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Sample records for chemically-induced rat mesenchymal

  1. Comparative sensitivity of human and rat neural cultures to chemical-induced inhibition of neurite outgrowth

    Energy Technology Data Exchange (ETDEWEB)

    Harrill, Joshua A.; Freudenrich, Theresa M.; Robinette, Brian L.; Mundy, William R., E-mail: mundy.william@epa.gov

    2011-11-15

    cultures were more sensitive to neurite outgrowth inhibitors, they also had a lower dynamic range for detecting chemical-induced neurite outgrowth inhibition and greater variability from culture-to-culture as compared to rat primary cortical cultures.

  2. Comparative sensitivity of human and rat neural cultures to chemical-induced inhibition of neurite outgrowth

    International Nuclear Information System (INIS)

    cultures were more sensitive to neurite outgrowth inhibitors, they also had a lower dynamic range for detecting chemical-induced neurite outgrowth inhibition and greater variability from culture-to-culture as compared to rat primary cortical cultures.

  3. Use of a chemically induced-colon carcinogenesis-prone Apc-mutant rat in a chemotherapeutic bioassay

    International Nuclear Information System (INIS)

    Chemotherapeutic bioassay for colorectal cancer (CRC) with a rat model bearing chemically-induced CRCs plays an important role in the development of new anti-tumor drugs and regimens. Although several protocols to induce CRCs have been developed, the incidence and number of CRCs are not much enough for the efficient bioassay. Recently, we established the very efficient system to induce CRCs with a chemically induced-colon carcinogenesis-prone Apc-mutant rat, Kyoto Apc Delta (KAD) rat. Here, we applied the KAD rat to the chemotherapeutic bioassay for CRC and showed the utility of the KAD rat. The KAD rat has been developed by the ENU mutagenesis and carries a homozygous nonsense mutation in the Apc gene (S2523X). Male KAD rats were given a single subcutaneous injection of AOM (20 mg/kg body weight) at 5 weeks of age. Starting at 1 week after the AOM injection, they were given 2% DSS in drinking water for 7 days. Tumor-bearing KAD rats were divided into experimental and control groups on the basis of the number of tumors observed by endoscopy at week 8. The 5-fluorouracil (5-FU) was administrated intravenously a dose of 50 or 75 mg/kg weekly at week 9, 10, and 11. After one-week interval, the 5-FU was given again at week 13, 14, and 15. At week 16, animals were sacrificed and tumor number and volume were measured macroscopically and microscopically. In total 48 tumors were observed in 27 KAD rats with a 100% incidence at week 8. The maximum tolerated dose for the KAD rat was 50 mg/kg of 5-FU. Macroscopically, the number or volume of tumors in the 5-FU treated rats was not significantly different from the control. Microscopically, the number of adenocarcinoma in the 5-FU treated rats was not significantly different (p < 0.02) from that of the control. However, the volume of adenocarcinomas was significantly lower than in the control. Anticancer effect of the 5-FU could be obtained only after the 16 weeks of experimental period. The use of the AOM/DSS-treated tumor

  4. Saffron Aqueous Extract Inhibits the Chemically-induced Gastric Cancer Progression in the Wistar Albino Rat

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    S. Zahra Bathaie

    2013-01-01

    Full Text Available Objective(s: Gastric cancer is the first and second leading cause of cancer related death in Iranian men and women, respectively. Gastric cancer management is based on the surgery, radiotherapy and chemotherapy. In the present study, for the first time, the beneficial effect of saffron (Crocus sativus L. aqueous extract (SAE on the 1-Methyl-3-nitro-1-nitrosoguanidine (MNNG-induced gastric cancer in rat was investigated. Materials and Methods: MNNG was used to induce gastric cancer and then, different concentrations of SAE were administered to rats. After sacrificing, the stomach tissue was investigated by both pathologist and flow cytometry, and several biochemical parameters was determined in the plasma (or serum and stomach of rats. Results: Pathologic data indicated the induction of cancer at different stages from hyperplasia to adenoma in rats; and the inhibition of cancer progression in the gastric tissue by SAE administration; so that, 20% of cancerous rats treated with higher doses of SAE was completely normal at the end of experiment and there was no rat with adenoma in the SAE treated groups. In addition, the results of the flow cytometry/ propidium iodide staining showed that the apoptosis/proliferation ratio was increased due to the SAE treatment of cancerous rats. Moreover, the significantly increased serum LDH and decreased plasma antioxidant activity due to cancer induction fell backwards after treatment of rats with SAE. But changes in the other parameters (Ca2+, tyrosine kinase activity and carcino-embryonic antigen were not significant. Conclusion: SAE inhibits the progression of gastric cancer in rats, in a dose dependent manner.

  5. Aqueous suspension of anise "Pimpinella anisum" protects rats against chemically induced gastric ulcers

    Institute of Scientific and Technical Information of China (English)

    Ibrahim A Al Mofleh; Abdulqader A Alhaider; Jaber S Mossa; Mohammed O Al-Soohaibani; Syed Rafatullah

    2007-01-01

    AIM:To substantiate the claims of Unani and Arabian traditional medicine practitioners on the gastroprotective potential effect of a popular spice anise,"Pimpinella anisum L." on experimentally-induced gastric ulceration and secretion in rats.METHODS:Acute gastric ulceration in rats was produced by various noxious chemicals including 80% ethanol,0.2 mol/L NaOH,25% NaCl and indomethacin.Anti-secretory studies were undertaken using pylorusligated Shay rat technique.Levels of gastric non-protein sulfhydryls(NP-SH)and wall mucus were estimated and gastric tissue was also examined histologically.Anise aqueous suspension was used in two doses(250 and 500 mg/kg body weight)in all experiments.RESULTS:Anise significantly inhibited gastric mucosal damage induced by necrotizing agents and indomethacin.The anti-ulcer effect was further confirmed histologically.In pylorus-ligated Shay rats,anise suspension significantly reduced the basal gastric acid secretion,acidity and completely inhibited the rumenal ulceration.On the other hand,the suspension significantly replenished ethanol-induced depleted levels of gastric mucosal NP-SH and gastric wall mucus concentration.CONCLUSION:Anise aqueous suspension possesses significant cytoprotective and anti-ulcer activities against experimentally-induced gastric lesions.The anti-ulcer effect of anise is possibly prostaglandin-mediated and/or through its anti-secretory and antioxidative properties.

  6. Efficacy of Mesenchymal Stem Cells in Suppression of Hepatocarcinorigenesis in Rats: Possible Role of Wnt Signaling

    LENUS (Irish Health Repository)

    Abdel Aziz, Mohamed T

    2011-05-05

    Abstract Background The present study was conducted to evaluate the tumor suppressive effects of bone marrow derived mesenchymal stem cells (MSCs) in an experimental hepatocellular carcinoma (HCC) model in rats and to investigate the possible role of Wnt signaling in hepato-carcinogenesis. Methods Ninety rats were included in the study and were divided equally into: Control group, rats which received MSCs only, rats which received MSCs vehicle only, HCC group induced by diethylnitroseamine (DENA) and CCl 4 , rats which received MSCs after HCC induction, rats which received MSCs before HCC induction. Histopathological examination and gene expression of Wnt signaling target genes by real time, reverse transcription-polymerase chain reaction (RT-PCR) in rat liver tissue, in addition to serum levels of ALT, AST and alpha fetoprotein were performed in all groups. Results Histopathological examination of liver tissue from animals which received DENA-CCl4 only, revealed the presence of anaplastic carcinoma cells and macro-regenerative nodules type II with foci of large and small cell dysplasia. Administration of MSCs into rats after induction of experimental HCC improved the histopathological picture which showed minimal liver cell damage, reversible changes, areas of cell drop out filled with stem cells. Gene expression in rat liver tissue demonstrated that MSCs downregulated β-catenin, proliferating cell nuclear antigen (PCNA), cyclin D and survivin genes expression in liver tissues after HCC induction. Amelioration of the liver status after administration of MSCs has been inferred by the significant decrease of ALT, AST and Alpha fetoprotein serum levels. Administration of MSCs before HCC induction did not show any tumor suppressive or protective effect. Conclusions Administration of MSCs in chemically induced HCC has tumor suppressive effects as evidenced by down regulation of Wnt signaling target genes concerned with antiapoptosis, mitogenesis, cell proliferation

  7. Effect of chronic administration of green tea extract on chemically induced electrocardiographic and biochemical changes in rat heart

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    Patil Leena

    2010-01-01

    Full Text Available Many chemicals induce cell-specific cytotoxicity. Chemicals like doxorubicin induce oxidative stress leading to cardiotoxicity causing abnormalities in ECG and increase in the biomarkers indicating toxicity. Green tea extract (GTE, Camellia sinensis (Theaceae, is reported to exert antioxidant activity mainly by means of its polyphenolic constituent, catechins. Our study was aimed to find out the effect of GTE (25, 50, 100 mg/kg/day p.o. for 30 days on doxorubicin-induced (3 mg/kg/week, i.p. for 5 weeks electrocardiographic and biochemical changes in rat heart. It is observed that GTE administered rats were less susceptible to doxorubicin-induced electrocardiographic changes and changes in biochemical markers like lactate dehydrogenase (LDH, creatine kinase (CK, and glutamic oxaloacetate transaminase (GOT in serum, and superoxide dismutase (SOD, catalase (CAT and reduced glutathione (GSH, membrane bound enzymes like Na + K + ATPase, Ca 2+ ATPase, Mg 2+ ATPase and decreased lipid peroxidation (LP in heart tissue, indicating the protection afforded by GTE administration.

  8. Extract of the Bark of Bathysa cuspidataAttenuates the Development of Chemically-Induced Preneoplastic Colorectal Lesions in Rats

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    Damiana Diniz Rosa

    2015-10-01

    Full Text Available ABSTRACTThe aim of this study was to investigate the effect of the bark extractBathysa cuspidata on chemically induced preneoplastic colorectal lesions in Wistar rats. Forty male rats were randomly divided into four groups (n = 10 each: saline (control group, oral administration of saline solution 0.9%; dimethylsulfoxide (DMSO, vehicle control, B200 (treated with 200 mg/kg bark extract ofB. cuspidata, and B400 (treated with 400 mg/kg bark extract ofB. cuspidata. Administration of treatments was carried out by the gavage. The animals received four subcutaneous injections of 1,2-dimethylhydrazine (DMH, 40 mg/kg in the initial two weeks of the experiment to induce preneoplastic colorectal lesions. After 15 weeks, the animals were euthanized and the presence of aberrant crypt foci (ACF, body weight, biochemical analyses, and oxidative stress markers were measured. The extract ofB. cuspidata decreased the levels of superoxide dismutase (SOD, but did not influence the levels of catalase (CAT, malondialdehyde (MDA, nitric oxide or protein carbonyl, compared with the saline group. The animals supplemented with a more concentratedB. cuspidata extract (B400 showed a significant reduction in the number of ACF in all the portions of the intestinal mucosa. The study demonstrated that the bark extract ofB. cuspidata at 400 mg/kg reduced the preneoplastic colorectal lesions in an animal model of colon cancer and that the effect could be dose-dependent.

  9. Therapeutic Efficacy of Ginger, Cisplatin and Radiation on Chemically-Induced Cancer in Male Albino Rats

    International Nuclear Information System (INIS)

    This study aimed to investigate the in vivo effect of dietary supplementation with ginger to evaluate its therapeutic effect against lung and kidney cancer and in combination with cisplatin as chemotherapy and radiotherapy in male albino rats. 54 male albino rats were divided into nine groups of 6 animals each, all animals were allowed to food and water ad libitum . Group I was treated with 0.5 ml saline, orlly for 12 consecutive weeks serve as con - trol group Group II injected with N-nitrosodimethylamine (NDMA) and carbon tetrachloride (CCl4 ); all groups were injected with NDMA + CCl4 for 6 weeks. Group III were given ginger for 6 consecutive weeks (200 mg/kg, b.wt./day). Group IV animals received cisplatin, group V irradiated with 2 Gy, group VI treated with ginger then irradiated, group VII treated with ginger then injected with cisplatin, group VIII injected with cisplatin then irradiated and group IX treated with ginger and cisplatin then irradiated. Antioxidant status in both kidney and lung tissues were estimated by determining the activity of antioxidant enzyme superoxide dismutase (SOD); as well as the level of reduced glutathione (GSH), Malondialdehyde (MDA) and Nitric oxide (NO). In parallel to histopathological investigations of lung and kidney tissues. In addition, Tumor Necrosis Factor Alpha (TNF-α) level, advanced oxidative protein product (AOPP), urea, creatinine and uric acid. Remarkable disturbances were observed in the levels of all tested parameters in NDMA + CCl4 group. On the other hand, rats injected with the cancer agents then treated with cisplatin+radiation showed moderate improvements in the studied parameters while, treatment with ginger + cisplatin + radiation ameliorated the levels of the disturbed bio

  10. The effect of probiotic microorganisms and bioactive compounds on chemically induced carcinogenesis in rats.

    Science.gov (United States)

    Bertkova, I; Hijova, E; Chmelarova, A; Mojzisova, G; Petrasova, D; Strojny, L; Bomba, A; Zitnan, R

    2010-01-01

    Diet interventions and natural bioactive supplements have now been extensively studied to reduce risks of colon cancer, which is one of the major public health problem throughout the world. The objective of our investigation was to study the effects of probiotic, prebiotic, nutritional plant extract, and plant oil on selected biochemical and immunological parameters in rats with colon cancer induced by N,N dimethylhydrazine (DMH). Male and female Wistar albino rats were were fed by a high-fat (HF) diet (10% fat in the diet) and were divided into 9 groups: Control group; PRO group - HF diet supplemented with probiotic Lactobacillus plantarum to provide 3 x 109 c.f.u. of strain/1 ml of medium; PRE group - HF diet supplemented with inulin enriched with oligofructose (2% of HF diet); HES group - HF diet supplemented with plant extract of Aesculus hippocastanum L. (1% of HF diet); OIL group - HF diet comprised Linioleum virginale (2% of HF diet); and combination of probiotic microorganisms and bioactive compounds in the groups - PRO-PRE, PRO-HES, PRO-OIL, PRE-OIL. Carcinogenesis was initiated with subcutaneous injection of DMH (20 mg/kg) two times at week interval and dietary treatments were continued for the six weeks. Application of probiotic microorganisms and bioactive compounds in all treated groups significantly decreased the activities of bacterial enzymes (p<0.001), the fecal bile acids concentration (p<0.01; p<0.001) and significantly increased serum TNFalpha level (p<0.001) in comparison to the control rats. The number of coliforms was reduced in PRO, PRO-PRE, PRO-OIL and PRE-OIL groups and significantly higher count of lactobacilli (p<0.05) was observed in PRO-PRE, PRO-OIL and PRE-OIL groups in compare with the controls. In conclusion, the results of this study indicate that probiotic microorganisms and bioactive compounds could exert a preventive effect on colon carcinogenesis induced by DMH. PMID:20568896

  11. Chemically induced immunotoxicity in a medium-term multiorgan bioassay for carcinogenesis with Wistar rats

    International Nuclear Information System (INIS)

    A variety of chemicals can adversely affect the immune system and influence tumor development. The modifying potential of chemical carcinogens on the lymphoid organs and cytokine production of rats submitted to a medium-term initiation-promotion bioassay for carcinogenesis was investigated. Male Wistar rats were sequentially initiated with N-nitrosodiethylamine (DEN), N-methyl-N-nitrosourea (MNU), N-butyl-N-(4hydroxybutyl)nitrosamine (BBN), dihydroxy-di-n-propylnitrosamine (DHPN), and 1,2-dimethylhydrazine (DMH) during 4 weeks. Two initiated groups received phenobarbital (PB) or 2-acetylaminofluorene (2-AAF) for 25 weeks and two noninitiated groups received only PB or 2-AAF. A nontreated group was used as control. Lymphohematopoietic organs, liver, kidneys, lung, intestines, and Zymbal's gland were removed for histological analysis. Interleukin (IL)-2, IL-12, interferon gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), IL-10, and transforming growth factor beta1 (TGF-β1) levels were determined by ELISA in spleen cell culture supernatants. At the fourth week, exposure to the initiating carcinogens resulted in cell depletion of the thymus, spleen and bone marrow, and impairment of IL-2, IL-12, and IFN-γ production. However, at the 30th week, no important alterations were observed both in lymphoid organs and cytokine production in the different groups. The results indicate that the initiating carcinogens used in the present protocol exert toxic effects on the lymphoid organs and affect the production of cytokines at the initiation step of carcinogenesis. This early and reversible depression of the immune surveillance may contribute to the survival of initiated cells facilitating the development of future neoplasia

  12. Rat epidermal keratinocyte organotypic culture (ROC) as a model for chemically induced skin irritation testing

    International Nuclear Information System (INIS)

    The potential of rat epidermal keratinocyte (REK) organotypic culture (ROC) with proper stratum corneum barrier as a model for screening skin irritants was evaluated. The test chemicals were selected from ECETOC database (1995) and the observed in vitro irritation potential was compared to ECETOC in vivo primary irritation index (PII), to EU risk phrases, and to the harmonized OECD criteria. Chemicals were applied onto the stratum corneum surface of ROC for 30 min and samples were taken from the underlying medium at 4 and 8 h after exposure. Cell membrane integrity (determined by LDH assay) and pro-inflammatory effect (determined by IL-1α release) were verified at both time points and correlated to PII values. The best correlation (R 2 = 0.831) was seen with LDH leakage test. Based on obtained data, chemicals were classified according to criteria defined by EU and OECD. From 12 chemicals, only two were incorrectly classified according to OECD criteria when using LDH leakage and IL-1α release as irritation markers. At the end of experiment, chemical-treated ROC cultures were fixed and histological changes were assessed. Typical signs for irritation were lightly stained cytoplasm, condensed nuclei, cellular vacuolization, eosinophilic cytoplasms, and blebbing. These irritation effects of chemicals were graded visually into four classes (A-D). The extent of morphological perturbations of the cultures mostly correlated with PII. The present results indicate the validity of the ROC model in predicting skin irritation potential of chemicals and show that the use of set of irritation markers with different mechanistic responses gives more information on irritation than if only one marker was used

  13. Heterologous mesenchymal stem cells successfully treat femoral pseudarthrosis in rats

    OpenAIRE

    Ferreira Manoel; Silva Paulo; Alvarez Silva Lucas; Bonfim Danielle; Conilho Macedo Müller Lucas; Espósito Christiano; Schanaider Alberto

    2012-01-01

    Abstract Background This study evaluated the effectiveness of treating pseudarthrosis in rats by using bone marrow cell suspensions or cultures of bone marrow mesenchymal stromal cells Methods Thirty-eight specific pathogen-free (SPF) animals were randomly assigned to four groups: Group 1, Control, without surgical intervention; Group 2 (Placebo), experimental model of femoral pseudarthrosis treated only with saline solution; Group 3, experimental model of femoral pseudarthrosis treated with ...

  14. Hepatoprotective and antioxidative effects of total phenolics from Laggera pterodonta on chemical-induced injury in primary cultured neonatal rat hepatocytes.

    Science.gov (United States)

    Wu, Yihang; Yang, Leixiang; Wang, Fang; Wu, Xiumei; Zhou, Changxin; Shi, Shuyun; Mo, Jianxia; Zhao, Yu

    2007-08-01

    Although Laggera pterodonta as a folk medicine has been widely used for several centuries to ameliorate some inflammatory ailments as hepatitis in China, there have been no studies of the hepatoprotective and antioxidative effects of this plant. In this paper, the hepatoprotective effect of total phenolics from L. pterodonta (TPLP) against CCI4-, D-GalN-, TAA-, and t-BHP-induced injury was examined in primary cultured neonatal rat hepatocytes. TPLP inhibited the cellular leakage of two enzymes, hepatocyte ASAT and ALAT, caused by these chemicals and improved cell viability. Moreover, TPLP afforded much stronger protection than the reference drug silibinin. Meanwhile, DPPH and superoxide radicals scavenging activities of TPLP were also determined. The present investigation is the first to report chemical-induced injury model in primary cultured neonatal rat hepatocytes and provide evidence for the hepatoprotective and antioxidative effects of L. pterodonta. Neutralizing reactive oxygen species by nonenzymatic mechanisms may be one of main mechanisms of TPLP against chemical-induced hepatocyte injury. Furthermore, The total phenolic content of L. pterodonta and its main component type were quantified, and its principle components isochlorogenic acids were isolated and authenticated. These data support the folkloric uses of L. pterodonta in the treatment of hepatitis. PMID:17329003

  15. Therapeutic efficacy of amniotic membrane stem cells and adipose tissue stem cells in rats with chemically induced ovarian failure.

    Science.gov (United States)

    Fouad, Hanan; Sabry, Dina; Elsetohy, Khaled; Fathy, Naglaa

    2016-03-01

    The present study was conducted to compare between the therapeutic efficacies of human amniotic membrane-derived stem cells (hAM-MSCs) vs. adipose tissue derived stem cells (AD-MSCs) in cyclophosphamide (CTX)-induced ovarian failure in rats. Forty-eight adult female rats were included in the study; 10 rats were used as control group. Thirty-eight rats were injected with CTX to induce ovarian failure and divided into four groups: ovarian failure (IOF) (IOF group), IOF + phosphate buffer saline (PBS group), IOF + hAM-MSCs group and IOF + AD-MSCs group. Serum levels of FSH and estradiol (E2) were assessed. Histopathological examination of the ovarian tissues was performed and quantitative gene expressions of Oct-4, Stra8 and integrin beta-1 genes were conducted by quantitative real time PCR. Results showed that IOF and IOF + PBS rat groups exhibited decreased ovarian follicles, increased interstitial fibrosis with significant decrease of serum E2, significant increase serum FSH level and significant down-regulation of Stra8 and integrin beta-1. In hAM-MSCs and AD-MSCs rat groups, there were increased follicles and corpora with evident the presence of oocytes, significant increase in serum E2, significant decrease in serum FSH levels (in hAM-MSCs treated group only) and significant up-regulation of the three studied genes with higher levels in hAM-MSCs treated rats group when compared to AD-MSCs treated rats group. In Conclusion, administration of either hAM-derived MSCs or AD-MSCs exerts a significant therapeutic efficacy in chemotherapy induced ovarian insult in rats. hAM-MSCs exert higher therapeutic efficacy as compared to AD-MSCs. PMID:26966564

  16. Characterization of chemically induced ovarian carcinomas in an ethanol-preferring rat model: influence of long-term melatonin treatment.

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    Luiz Gustavo A Chuffa

    Full Text Available Ovarian cancer is the fourth most common cause of cancer deaths among women, and chronic alcoholism may exert co-carcinogenic effects. Because melatonin (mel has oncostatic properties, we aimed to investigate and characterize the chemical induction of ovarian tumors in a model of ethanol-preferring rats and to verify the influence of mel treatment on the overall features of these tumors. After rats were selected to receive ethanol (EtOH, they were surgically injected with 100 µg of 7,12-dimethyl-benz[a]anthracene (DMBA plus sesame oil directly under the left ovarian bursa. At 260 days old, half of the animals received i.p. injections of 200 µg mel/100 g b.w. for 60 days. Four experimental groups were established: Group C, rats bearing ovarian carcinomas (OC; Group C+EtOH, rats voluntarily consuming 10% (v/v EtOH and bearing OC; Group C+M, rats bearing OC and receiving mel; and Group C+EtOH+M, rats with OC consuming EtOH and receiving mel. Estrous cycle and nutritional parameters were evaluated, and anatomopathological analyses of the ovarian tumors were conducted. The incidence of ovarian tumors was higher in EtOH drinking animals 120 days post-DMBA administration, and mel efficiently reduced the prevalence of some aggressive tumors. Although mel promoted high EtOH consumption, it was effective in synchronizing the estrous cycle and reducing ovarian tumor mass by 20%. While rats in the C group displayed cysts containing serous fluid, C+EtOH rats showed solid tumor masses. After mel treatment, the ovaries of these rats presented as soft and mobile tissues. EtOH consumption increased the incidence of serous papillary carcinomas and sarcomas but not clear cell carcinomas. In contrast, mel reduced the incidence of sarcomas, endometrioid carcinomas and cystic teratomas. Combination of DMBA with EtOH intake potentiated the incidence of OC with malignant histologic subtypes. We concluded that mel reduces ovarian masses and the incidence of

  17. Enhancement of mouse germ cell-associated genes expression by injection of human umbilical cord mesenchymal stem cells into the testis of chemical-induced azoospermic mice

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    Rui-Feng Yang

    2014-10-01

    Full Text Available Various methods are currently under investigation to preserve fertility in males treated with high-dose chemotherapy and radiation for malignant and nonmalignant disorders. Human umbilical cord mesenchymal stem cells (HUC-MSCs, which possess potent immunosuppressive function and secrete various cytokines and growth factors, have the potential clinical applications. As a potential alternative, we investigate whether injection of HUC-MSCs into the interstitial compartment of the testes to promote spermatogenic regeneration efficiently. HUC-MSCs were isolated from different sources of umbilical cords and injected into the interstitial space of one testis from 10 busulfan-treated mice (saline and HEK293 cells injections were performed in a separate set of mice and the other testis remained uninjected. Three weeks after MSCs injection, Relative quantitative reverse transcription polymerase chain reaction was used to identify the expression of 10 of germ cell associated, which are all related to meiosis, demonstrated higher levels of spermatogenic gene expression (2-8 fold in HUC-MSCs injected testes compared to the contralateral uninjected testes (five mice. Protein levels for germ cell-specific genes, miwi, vasa and synaptonemal complex protein (Scp3 were also higher in MSC-treated testes compared to injected controls 3 weeks after treatment. However, no different expression was detected in saline water and HEK293 cells injection control group. We have demonstrated HUC-MSCs could affect mouse germ cell-specific genes expression. The results also provide a possibility that the transplanted HUC-MSCs may promote the recovery of spermatogenesis. This study provides further evidence for preclinical therapeutic effects of HUC-MSCs, and explores a new approach to the treatment of azoospermia.

  18. Graptopetalum paraguayense ameliorates chemical-induced rat hepatic fibrosis in vivo and inactivates stellate cells and Kupffer cells in vitro.

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    Li-Jen Su

    Full Text Available BACKGROUND: Graptopetalum paraguayense (GP is a folk herbal medicine with hepatoprotective effects that is used in Taiwan. The aim of this study was to evaluate the hepatoprotective and antifibrotic effects of GP on experimental hepatic fibrosis in both dimethylnitrosamine (DMN- and carbon tetrachloride (CCl(4-induced liver injury rats. METHODS: Hepatic fibrosis-induced rats were fed with the methanolic extract of GP (MGP by oral administration every day. Immunohistochemistry, biochemical assays, and Western blot analysis were performed. The effects of MGP on the expression of fibrotic markers and cytokines in the primary cultured hepatic stellate cells (HSCs and Kupffer cells, respectively, were evaluated. RESULTS: Oral administration of MGP significantly alleviated DMN- or CCl(4-induced liver inflammation and fibrosis. High levels of alanine transaminase, aspartate transaminase, bilirubin, prothrombin activity and mortality rates also decreased in rats treated with MGP. There were significantly decreased hydroxyproline levels in therapeutic rats compared with those of the liver-damaged rats. Collagen I and alpha smooth muscle actin (α-SMA expression were all reduced by incubation with MGP in primary cultured rat HSCs. Furthermore, MGP induced apoptotic cell death in activated HSCs. MGP also suppressed lipopolysaccharide-stimulated rat Kupffer cell activation by decreasing nitric oxide, tumor necrosis factor-α and interleukin-6 production, and increasing interleukin-10 expression. CONCLUSIONS: The results show that the administration of MGP attenuated toxin-induced hepatic damage and fibrosis in vivo and inhibited HSC and Kupffer cell activation in vitro, suggesting that MGP might be a promising complementary or alternative therapeutic agent for liver inflammation and fibrosis.

  19. QUANTITATIVE CHANGES IN THE SYNAPTIC VESICLE PROTEINS SYNAPSIN I AND P38 AND THE ASTROCYTE-SPECIFIC PROTEIN GLIAL FIBRILLARY ACIDIC PROTEIN ARE ASSOCIATED WITH CHEMICAL-INDUCED INJURY TO THE RAT CENTRAL NERVOUS SYSTEM (JOURNAL VERSION)

    Science.gov (United States)

    Measurements of neuron-specific and glia-specific proteins were used to characterize chemical-induced injury to the rat CNS. Trimethyltin (TMT), a neurotoxicant which preferentially damages neurons in limbic structures, was employed to produce consistent, time-dependent, dose-rel...

  20. The Possible Effect Of Tamoxifen Vs Whole Body Irradiation Treatment On Thyroid Hormones in Female Rats Bearing Mammary Tumors Chemically Induced

    International Nuclear Information System (INIS)

    Breast cancer is the most common malignancy among women in most developed and developing regions of the world. In women, this drug has tissuespecific effects, acting as an estrogen antagonist on the breast, and as an estrogen agonist on bone, lipid metabolism (increasing high-density lipoprotein cholesterol and decreasing low-density lipoprotein cholesterol), and the endometrium. Thyroid hormones act on almost all organs throughout the body and regulate the basal metabolism of the organism. Thyroid hormone can also stimulate the proliferation in vitro of certain tumor cell lines. The aim of the present study is to evaluate the significant value of tamoxifen and/or irradiation treatment on thyroid hormones in breast cancer bearing female rats. Forty two female Sprague-Dawely rats randomly divided into seven groups and the effect of tamoxifen and post-irradiation was studied on breast cancer chemically induced. The results shows a T4 and estradiol levels not T3 were altered in different experimental groups. It could be concluded that irradiation-induced changes in the composition of the mammary microenvironment promote the expression of neoplastic potential by affecting both estradiol and thyroid hormones, and tamoxifen may alter the thyroid hormones. Irradiation and tamoxifen administration may have worth effects on T4 and estradiol levels and it is recommended to further studies towards the bystander effect of radiation and tamoxifen on the tissue culture and molecular biology scale.

  1. Edible oils for liver protection: hepatoprotective potentiality of Moringa oleifera seed oil against chemical-induced hepatitis in rats.

    Science.gov (United States)

    Al-Said, Mansour S; Mothana, Ramzi A; Al-Yahya, Mohammed A; Al-Blowi, Ali S; Al-Sohaibani, Mohammed; Ahmed, Atallah F; Rafatullah, Syed

    2012-07-01

    In the present study, in vitro antioxidant, antioxidative stress and hepatoprotective activity of Moringa oleifera Lam. seed oil (Ben oil; BO) was evaluated against carbon tetrachloride (CCl(4) ) induced lipid peroxidation and hepatic damage in rats. The oil at 0.2 and 0.4 mL/rat was administered orally for 21 consecutive days. The substantially elevated serum enzymatic (GOT, GPT, ALP, GGT) and bilirubin levels were significantly restored towards normalization by the oil. There was a significant elevation in the level of malondialdehyde (MDA), non-protein sulfhydryl (NP-SH), and total protein (TP) contents in the liver tissue. The results obtained indicated that BO possesses potent hepatoprotective action against CCl(4) -induced hepatic damage by lowering liver marker enzymes, MDA concentration, and elevating NP-SH and TP levels in liver tissue. The biochemical observations were supplemented with histopathological examination of rat liver. The results of this study showed that treatment with Ben oil or silymarin (as a reference) appears to enhance the recovery from hepatic damage induced by CCl(4) . The pentobarbital induced narcolepsy prolongation in mice was retarded by the Ben oil. Acute toxicity test in mice showed no morbidity or mortality. In vitro DPPH radical scavenging and β-carotene-linolic acid assay tests of the BO exhibited a moderate antioxidant activity in both tests used. The possible mechanism(s) of the liver protective activity of Ben oil activity may be due to free radical scavenging potential caused by the presence of antioxidant component(s) in the oil. Consequently, BO can be used as a therapeutic regime in treatment of some hepatic disorders. PMID:22757719

  2. A robust method for assessing chemically induced mutagenic effects in the oral cavity of transgenic Big Blue® rats.

    Science.gov (United States)

    Young, Robert R; Thompson, Chad M; Dinesdurage, Harshini R; Elbekai, Reem H; Suh, Mina; Rohr, Annette C; Proctor, Deborah M

    2015-08-01

    The Big Blue® (BB) in vivo mutation assay uses transgenic rodents to measure treatment-induced mutations in virtually any tissue. The BB assay can be conducted in rats or mice and is ideal for investigating tissue-specific mutagenic mode of action of tumor induction. Some tissues such as oral mucosa have not been thoroughly studied. Due to the small quantity and cartilaginous nature of oral cavity tissues, development of special prosection and DNA isolation methods was required to permit robust analysis of mutations in these tissues. Improved surgical methods permitted collection of adequate and reproducible quantities of tissue (∼45 mg gingiva/buccal and ∼30 mg gingiva/palate). Optimized DNA isolation methods included use of liquid nitrogen pulverization, homogenization, nuclei pelleting, digestion, and phenol/chloroform extraction, to yield sufficient quantities of DNA from these tissues. In preliminary optimization work, mutant frequency (MF) in tongue and gingiva was increased in rats exposed to the promutagen, benzo[a]pyrene, and the direct mutagen, N-ethyl-N-nitrosourea. The oral cavity carcinogen, 4-nitroquinoline-1-oxide (4-NQO; 10 ppm in drinking water; 28 days), was qualified as a positive control for mutagenesis in oral tissues since it caused significant increases in cII MFs in gingiva/palate (50.2-fold) and gingiva/buccal tissues (21.3-fold), but not in liver or bone marrow (0.9- and 1.4-fold, respectively). These results are consistent with the observation that 4-NQO primarily induces tumors in oral cavity. Results also demonstrate the utility of the BB rat mutation assay and optimized methods for investigation of oral cavity mutagenicity, and by extension, analysis of other small and cartilaginous tissues. PMID:25969955

  3. M1-/M2-macrophages contribute to the development of GST-P-positive preneoplastic lesions in chemically-induced rat cirrhosis.

    Science.gov (United States)

    Wijesundera, Kavindra Kumara; Izawa, Takeshi; Tennakoon, Anusha Hemamali; Golbar, Hossain M; Tanaka, Miyuu; Kuwamura, Mitsuru; Yamate, Jyoji

    2015-09-01

    Glutathione S-transferase placental form (GST-P) expression in hepatocyte foci is regarded as a preneoplastic change in rats. We aimed to reveal the contribution of polarized macrophages in development of GST-P-positive pseudolobules (PLs) in chemically-induced rat cirrhosis. F344 rats were injected with thioacetamide (100mg/kg BW, twice a week, intraperitoneally). Macrophage immunophenotypes and expression of M1-/M2-related factors were analyzed by immunohistochemistry, real-time RT-PCR and laser microdissection. GST-P-positive foci/clusters were clearly observed at post-first injection week 15. GST-P-positive PLs were distinguishable at weeks 20-32. Microarray analysis revealed upregulation of preneoplastic genes in GST-P-positive PLs at week 32. M1 (CD68(+), Iba1(+))-and M2 (CD163(+), CD204(+), Gal-3(+))-macrophages were greater in number in the GST-P-positive PLs, whereas MHC class II-positive (M1) macrophage number was fewer in the GST-P-positive PLs. Expression of both M1 (IFN-γ, IL-1β, TNF-α, Iba1)- and M2 (IL-4, TGF-β1, IL-10)-related factors were higher in GST-P-positive PLs. Our results showed that both M1- and M2-macrophage populations contribute to the development of hepatic preneoplastic lesions. MHC class II-positive macrophages may be related to anti-tumor progression, since their kinetics showed reverse pattern to other macrophage phenotypes. PMID:26205097

  4. Isolation, culturing and characterization of rat adipose tissue-derived mesenchymal stem cells: a simple technique

    OpenAIRE

    NİYAZ, Mehmet; Özer Aylin GÜRPINAR; GÜNAYDIN, Serdar; Onur, Mehmet Ali

    2012-01-01

    In this study, our aim was to develop a new simple technique for isolation of mesenchymal stem cells from adipose tissue. For this purpose, mesenchymal stem cells were isolated from rat adipose tissue by using the primary explant culture technique. When the cells became confluent, they were passaged 4 times by using the standard trypsinization method with trypsin/EDTA solution. Cells at second passage were characterized by using immunofluorescence staining against CD13 and CD29 markers. The r...

  5. Pelvic Organ Distribution of Mesenchymal Stem Cells Injected Intravenously after Simulated Childbirth Injury in Female Rats

    OpenAIRE

    Michelle Cruz; Charuspong Dissaranan; Anne Cotleur; Matthew Kiedrowski; Marc Penn; Margot Damaser

    2011-01-01

    The local route of stem cell administration utilized presently in clinical trials for stress incontinence may not take full advantage of the capabilities of these cells. The goal of this study was to evaluate if intravenously injected mesenchymal stem cells (MSCs) home to pelvic organs after simulated childbirth injury in a rat model. Female rats underwent either vaginal distension (VD) or sham VD. All rats received 2 million GFP-labeled MSCs intravenously 1 hour after injury. Four or 10 days...

  6. Reconstruction of the corneal epithelium with induced marrow mesenchymal stem cells in rats

    OpenAIRE

    Jiang, Ting-Shuai; Cai, Li; Ji, Wei-Ying; Hui, Yan-Nian; Wang, Yu-Sheng; Hu, Dan; Zhu, Jie

    2010-01-01

    Purpose To explore the feasibility of bone marrow mesenchymal stem cells (MSCs) transdifferentiating into corneal epithelial cells in a limbal stem cell deficiency (LSCD) model in rats. Methods Rat MSCs were isolated and purified using a gradient isolation procedure. The cells were induced by rat corneal stromal cells (CSCs) in a transwell co-culture system. The induced MSCs were identified by immunofluorescence staining, flow cytometry, and scanning electron microscopy (SEM). A corneal LSCD ...

  7. Use of FK506 and bone marrow mesenchymal stem cells for rat hind limb allografts

    Institute of Scientific and Technical Information of China (English)

    Youxin Song; Zhujun Wang; Zhixue Wang; Hong Zhang; Xiaohui Li; Bin Chen

    2012-01-01

    Dark Agouti rat donor hind limbs were orthotopically transplanted into Lewis rat recipients to verify the effects of bone marrow mesenchymal stem cells on neural regeneration and functional recovery of allotransplanted limbs in the microenvironment of immunotolerance. bone marrow mesenchymal stem cells were intramuscularly (gluteus maximus) injected with FK506 (tacrolimus) daily, and were transplanted to the injured nerves. Results indicated that the allograft group not receiving therapy showed severe rejection, with transplanted limbs detaching at 10 days after transplantation with complete necrosis. The number of myelinated axons and Schwann cells in the FK506 and FK506 + bone marrow mesenchymal stem cells groups were significantly increased. We observed a lesser degree of gastrocnemius muscle degeneration, and increased polymorphic fibers along with other pathological changes in the FK506 + bone marrow mesenchymal stem cells group. The FK506 + bone marrow mesenchymal stem cells group showed significantly better recovery than the autograft and FK506 groups. The results demonstrated that FK506 improved the immune microenvironment. FK506 combined with bone marrow mesenchymal stem cells significantly promoted sciatic nerve regeneration, and improved sensory recovery and motor function in hind limb allotransplant.

  8. Glucocorticoids induce autophagy in rat bone marrow mesenchymal stem cells

    DEFF Research Database (Denmark)

    Wang, L.; Fan, J.; Lin, Y. S.;

    2015-01-01

    and their responses to diverse stimuli, however, the role of autophagy in glucocorticoidinduced damage to bone marrow mesenchymal stem cells (BMSCs) remains unclear. The current study confirmed that glucocorticoid administration impaired the proliferation of BMSCs. Transmission electron microscopy...

  9. Bone marrow mesenchymal stem cells combined with minocycline improve spinal cord injury in a rat model

    OpenAIRE

    Chen, Dayong; Zeng, Wei; Fu, Yunfeng; Gao, Meng; Lv, Guohua

    2015-01-01

    The aims of this study were to assess that the effects of bone marrow mesenchymal stem cells (BMSCs) combination with minocycline improve spinal cord injury (SCI) in rat model. In the present study, the Wistar rats were randomly divided into five groups: control group, SCI group, BMSCs group, Minocycline group and BMSCs + minocycline group. Basso, Beattie and Bresnahan (BBB) test and MPO activity were used to assess the effect of combination therapy on locomotion and neutrophil infiltration. ...

  10. Spinal cord injury in rats treated using bone marrow mesenchymal stem-cell transplantation

    OpenAIRE

    Chen, Yu-Bing; Jia, Quan-Zhang; Li, Dong-Jun; Sun, Jing-Hai; Xi, Shuang; Liu, Li-ping; Gao, De-Xuan; Jiang, Da-Wei

    2015-01-01

    The aim of this study was to observe the effects of bone marrow mesenchymal stem-cell transplantation (BMSCs) in repairing acute spinal cord damage in rats and to examine the potential beneficial effects. 192 Wistar rats were randomized into 8 groups. Spinal cord injury was created. Behavior and limb functions were scored. Repairing effects of BMSCs transplantation was evaluated and compared. In vitro 4’,6-diamidino-2-phenylindole (DAPI)-tagged BMSCs were observed, and whether they migrated t...

  11. Lymphatic spread of mesenchymal renal tumor to metastatic parathymic lymph nodes in rat

    OpenAIRE

    Rozsa, David; Trencsenyi, Gyorgy; Kertai, Pal; Marian, Terez; Nagy, Gabor; Banfalvi, Gaspar

    2009-01-01

    Rat mesenchymal renal tumor cells (NeDe) transplanted under the kidney capsule of F344 rats resulted in metastases in the parathymic lymph nodes. Tumor cells were isolated from these tumor-bearing lymph nodes and 106 cells were implanted under the kidney capsule. Tumor growth after this implantation could be traced within six days. India ink was implanted to prove that there is a connection between the lymphatic vessels of the kidney capsule and the parathymic lymph ...

  12. Human bone marrow mesenchymal stem cell transplantation attenuates axonal injury in stroke rats

    OpenAIRE

    Xu, Yi; Du, Shiwei; Yu, Xinguang; HAN, XIAO; Hou, Jincai; Guo, Hao

    2014-01-01

    Previous studies have shown that transplantation of human bone marrow mesenchymal stem cells promotes neural functional recovery after stroke, but the neurorestorative mechanisms remain largely unknown. We hypothesized that functional recovery of myelinated axons may be one of underlying mechanisms. In this study, an ischemia/reperfusion rat model was established using the middle cerebral artery occlusion method. Rats were used to test the hypothesis that intravenous transplantation of human ...

  13. Human bone marrow mesenchymal stem cell transplantation attenuates axonal injur y in stroke rats

    Institute of Scientific and Technical Information of China (English)

    Yi Xu; Shiwei Du; Xinguang Yu; Xiao Han; Jincai Hou; Hao Guo

    2014-01-01

    Previous studies have shown that transplantation of human bone marrow mesenchymal stem cells promotes neural functional recovery after stroke, but the neurorestorative mechanisms remain largely unknown. We hypothesized that functional recovery of myelinated axons may be one of underlying mechanisms. In this study, an ischemia/reperfusion rat model was established using the middle cerebral artery occlusion method. Rats were used to test the hypothesis that in-travenous transplantation of human bone marrow mesenchymal stem cells through the femoral vein could exert neuroprotective effects against cerebral ischemia via a mechanism associated with the ability to attenuate axonal injury. The results of behavioral tests, infarction volume analysis and immunohistochemistry showed that cerebral ischemia caused severe damage to the myelin sheath and axons. After rats were intravenously transplanted with human bone marrow mesenchymal stem cells, the levels of axon and myelin sheath-related proteins, including mi-crotubule-associated protein 2, myelin basic protein, and growth-associated protein 43, were elevated, infarct volume was decreased and neural function was improved in cerebral ischemic rats. These ifndings suggest that intravenously transplanted human bone marrow mesenchymal stem cells promote neural function. Possible mechanisms underlying these beneifcial effects in-clude resistance to demyelination after cerebral ischemia, prevention of axonal degeneration, and promotion of axonal regeneration.

  14. Effect of intravenous transplantation of bone marrow mesenchymal stem cells on neurotransmitters and synapsins in rats with spinal cord injury

    OpenAIRE

    Chen, Shaoqiang; Wu, Bilian; Lin, Jianhua

    2012-01-01

    Bone marrow mesenchymal stem cells were isolated, purified and cultured in vitro by Percoll density gradient centrifugation combined with the cell adherence method. Passages 3–5 bone marrow mesenchymal stem cells were transplanted into rats with traumatic spinal cord injury via the caudal vein. Basso-Beattie-Bresnahan scores indicate that neurological function of experimental rats was significantly improved over transplantation time (1–5 weeks). Expressions of choline acetyltransferase, gluta...

  15. Autologous adipose tissue-derived mesenchymal stem cells are involved in rat liver regeneration following repeat partial hepatectomy

    OpenAIRE

    Liu, Tao; Mu, Hong; SHEN, ZHONGYANG; Song, Zhuolun; CHEN, XIAOBO; Wang, Yuliang

    2016-01-01

    Adipose tissue-derived mesenchymal stem cells (ADSCs) have been considered to be attractive and readily available adult mesenchymal stem cells, and they are becoming increasingly popular for use in regenerative cell therapy, as they are readily accessible through minimally invasive techniques. The present study investigated whether autologous ADSC transplantation promoted liver regeneration following a repeat partial hepatectomy in rats. The rats were divided into three groups as follows: 70%...

  16. AB114. Myogenic differentiation of rat bone mesenchymal stem cells in vitro

    Science.gov (United States)

    Xiao, Heng-Jun; Yan, Wei-Xin; Chen, Jun; Gao, Xin; Zhuan, Li; Zang, Guang-Hui; Yang, Jun; Zhang, Yan; Liu, Ji-Hong

    2016-01-01

    Objective To culture and myogenic differentiation of rat bone mesenchymal stem cells (BMSCs) in vitro, and provide the available seed cells for erectile dysfunction (ED) therapy. Methods Rat BMSCs were isolated and cultured from the femur and tibia of Sprague Dawley (SD) rat. Mesenchymal stem cell positive cellular markers CD49d, CD73, CD90, CD105, CD106 and negative cellular markers CD31, CD34 and CD45 were arrayed by flow cytometry analysis. Furthermore, the fourth passage cells were induced and identified by their capacities in the myogenic differentiation. Results The cultured cells expressed mesenchymal stem cell positive cellular markers CD49d, CD73, CD90, CD105 and CD106, and lacked negative cellular markers CD31, CD34 and CD45. Myogenic differentiation cells can be stained with alpha-smooth muscle actin (α-SMA) and desmin, respectively. Conclusions Rat BMSCs have been successfully isolated, cultured and myogenic differentiation in vitro. They could be used as autogenous BMSCs and gene modified BMSCs for ED therapy.

  17. Therapeutic effect of bone marrow mesenchymal stem cells on cold stress induced changes in the hippocampus of rats

    OpenAIRE

    Kumar, Saravana Kumar Sampath; Perumal, Saraswathi; Rajagopalan, Vijayaraghavan

    2014-01-01

    The present study aims to evaluate the effect of bone marrow mesenchymal stem cells on cold stress induced neuronal changes in hippocampal CA1 region of Wistar rats. Bone marrow mesenchymal stem cells were isolated from a 6-week-old Wistar rat. Bone marrow from adult femora and tibia was collected and mesenchymal stem cells were cultured in minimal essential medium containing 10% heat-inactivated fetal bovine serum and were sub-cultured. Passage 3 cells were analyzed by flow cytometry for pos...

  18. Infusion of Mesenchymal Stem Cells Ameliorates Hyperglycemia in Type 2 Diabetic Rats

    OpenAIRE

    Si, Yiling; Zhao, Yali; Hao, Haojie; Liu, Jiejie; Guo, Yelei; Mu, Yiming; Shen, Jing; Cheng, Yu; Fu, Xiaobing; Han, Weidong

    2012-01-01

    Infusion of mesenchymal stem cells (MSCs) has been shown to effectively lower blood glucose in diabetic individuals, but the mechanism involved could not be adequately explained by their potential role in promoting islet regeneration. We therefore hypothesized that infused MSCs might also contribute to amelioration of the insulin resistance of peripheral insulin target tissues. To test the hypothesis, we induced a diabetic rat model by high-fat diet/streptozotocin (STZ) administration, perfor...

  19. Therapeutic effect of bone marrow mesenchymal stem cells on cold stress induced changes in the hippocampus of rats

    Institute of Scientific and Technical Information of China (English)

    Saravana Kumar Sampath Kumar; Saraswathi Perumal; Vijayaraghavan Rajagopalan

    2014-01-01

    The present study aims to evaluate the effect of bone marrow mesenchymal stem cells on cold stress induced neuronal changes in hippocampal CA1 region of Wistar rats. Bone marrow mes-enchymal stem cells were isolated from a 6-week-old Wistar rat. Bone marrow from adult femora and tibia was collected and mesenchymal stem cells were cultured in minimal essential medium containing 10% heat-inactivated fetal bovine serum and were sub-cultured. Passage 3 cells were analyzed by lfow cytometry for positive expression of CD44 and CD90 and negative expression of CD45. Once CD44 and CD90 positive expression was achieved, the cells were cultured again to 90% conlfuence for later experiments. Twenty-four rats aged 8 weeks old were randomly and evenly divided into normal control, cold water swim stress (cold stress), cold stress + PBS (intra-venous infusion), and cold stress + bone marrow mesenchymal stem cells (1 × 106; intravenous infusion) groups. The total period of study was 60 days which included 1 month stress period followed by 1 month treatment. Behavioral functional test was performed during the entire study period. After treatment, rats were sacriifced for histological studies. Treatment with bone marrow mesenchymal stem cells signiifcantly increased the number of neuronal cells in hippocampal CA1 region. Adult bone marrow mesenchymal stem cells injected by intravenous administration show potential therapeutic effects in cognitive decline associated with stress-related lesions.

  20. Transplanted Bone Marrow Mesenchymal Stem Cells Improve Memory in Rat Models of Alzheimer's Disease

    Directory of Open Access Journals (Sweden)

    Parvin Babaei

    2012-01-01

    Full Text Available The present study aims to evaluate the effect of bone marrow mesenchymal stem cells (MSCs grafts on cognition deficit in chemically and age-induced Alzheimer's models of rats. In the first experiments aged animals (30 months were tested in Morris water maze (MWM and divided into two groups: impaired memory and unimpaired memory. Impaired groups were divided into two groups and cannulated bilaterally at the CA1 of the hippocampus for delivery of mesenchymal stem cells (500×103/ and PBS (phosphate buffer saline. In the second experiment, Ibotenic acid (Ibo was injected bilaterally into the nucleus basalis magnocellularis (NBM of young rats (3 months and animals were tested in MWM. Then, animals with memory impairment received the following treatments: MSCs (500×103/ and PBS. Two months after the treatments, cognitive recovery was assessed by MWM in relearning paradigm in both experiments. Results showed that MSCs treatment significantly increased learning ability and memory in both age- and Ibo-induced memory impairment. Adult bone marrow mesenchymal stem cells show promise in treating cognitive decline associated with aging and NBM lesions.

  1. Human umbilical cord mesenchymal stem cells reduce systemic inflammation and attenuate LPS-induced acute lung injury in rats

    OpenAIRE

    Li Jianjun; Li Dong; Liu Xiaomei; Tang Shuhai; Wei Fengcai

    2012-01-01

    Abstract Background Mesenchymal stem cells (MSCs) possess potent immunomodulatory properties and simultaneously lack the ability to illicit immune responses. Hence, MSCs have emerged as a promising candidate for cellular therapeutics for inflammatory diseases. Within the context of this study, we investigated whether human umbilical cord-derived mesenchymal stem cells (UC-MSCs) could ameliorate lipopolysaccharide- (LPS-) induced acute lung injury (ALI) in a rat model. Methods ALI was induced ...

  2. Epithelial growth by rat vibrissae follicles in vitro requires mesenchymal contact via native extracellular matrix

    International Nuclear Information System (INIS)

    An in vitro assay utilizing the rat vibrissa anagen follicle as a model for studying the epithelial-mesenchymal interactions (EMI) in hair growth is described. Through selective disruption of the epithelial-mesenchymal interface, we investigate whether the specialized extracellular matrix (ECM) of the dermal papilla and basement membrane zone (BMZ) serves a crucial function in hair follicle EMI. Epithelial bulbs incubated intact within their follicular sheaths incorporate thymidine primarily into cells of the hair matrix and outer root sheath, as shown by autoradiography. However, after removal of its mesenchymal associations (dermal papilla and extrabulbar connective tissue), the epithelial bulb showed no incorporation. Neither externally added collagen (type I or IV) nor the basement membrane components in Matrigel could substitute for the growth supporting influence of native surrounding stroma. Mechanical separation of the bulb from the dermal papilla in the basement membrane zone inhibited thymidine incorporation by the epithelium even though mesenchyme was still in close proximity. Enzymatic digestion of the dermal papilla ECM and the basal lamina by Dispase, a fibronectinase and type IV collagenase, also inhibited bulb growth without evidence of cytotoxicity. These experiments suggest that direct epithelial to mesenchymal contact is required for the support of follicular epithelial growth in vitro and that specific ECM components, possibly fibronectin and/or type IV collagen, rather than diffusable factors alone, play a crucial role in the mechanism of hair follicle EMI. The in vitro system described here provides an alternative to developmental EMI models and may serve as a valuable tool for studying EMI in the adult mammalian organism

  3. Comparative study of the differentiation potential of rat bone marrow mesenchymal stem cells and rat muscle-derived stem cells

    Directory of Open Access Journals (Sweden)

    Ivan Alexandra

    2013-01-01

    Full Text Available We present a comparative study of the plasticity of rat bone marrow mesenchymal stem cells (MSCs and rat muscle-derived stem cells (MDSCs. The study was performed on two cell populations that were isolated by aspiration from the femur bone marrow and gastrocnemius muscle biopsy of 6-week-old albino rats. Both cell populations were exposed to identical stimulation conditions. The cells were capable of undergoing osteogenic, chondrogenic, adipogenic and epithelial differentiation, as shown by histochemistry and immunostaining techniques. The MDSC population showed behavior and characteristics similar to the bone marrow MSC population; however, the osteogenic and adipogenic potential was more reduced compared to MSCs. Our results indicate a positive expression of E cadherin and Cytokeratin 10 after 28 days under epithelial stimulation, suggesting a potential use for gastrocnemius muscle MDSCs as a promising source for regenerative therapies, including re-epithelialization and skin regeneration.

  4. Pelvic Organ Distribution of Mesenchymal Stem Cells Injected Intravenously after Simulated Childbirth Injury in Female Rats

    Directory of Open Access Journals (Sweden)

    Michelle Cruz

    2012-01-01

    Full Text Available The local route of stem cell administration utilized presently in clinical trials for stress incontinence may not take full advantage of the capabilities of these cells. The goal of this study was to evaluate if intravenously injected mesenchymal stem cells (MSCs home to pelvic organs after simulated childbirth injury in a rat model. Female rats underwent either vaginal distension (VD or sham VD. All rats received 2 million GFP-labeled MSCs intravenously 1 hour after injury. Four or 10 days later pelvic organs and muscles were imaged for visualization of GFP-positive cells. Significantly more MSCs home to the urethra, vagina, rectum, and levator ani muscle 4 days after VD than after sham VD. MSCs were present 10 days after injection but GFP intensity had decreased. This study provides basic science evidence that intravenous administration of MSCs could provide an effective route for cell-based therapy to facilitate repair after injury and treat stress incontinence.

  5. Bone marrow mesenchymal stem cells protect against retinal ganglion cell loss in aged rats with glaucoma

    Directory of Open Access Journals (Sweden)

    Hu Y

    2013-10-01

    Full Text Available Ying Hu,1,2 Hai Bo Tan,1 Xin Mei Wang,3 Hua Rong,1 Hong Ping Cui,1 Hao Cui2 Departments of Ophthalmology, 1Shanghai East Hospital of Tongji University, Shanghai, 2First Affiliated Hospital, 3Fourth Affiliated Hospital, Harbin Medical University, Harbin, People's Republic of China Abstract: Glaucoma is a common eye disease in the aged population and has severe consequences. The present study examined the therapeutic effects of bone marrow mesenchymal stem cell (BMSC transplantation in preventing loss of visual function in aged rats with glaucoma caused by laser-induced ocular hypertension. We found that BMSCs promoted survival of retinal ganglion cells in the transplanted eye as compared with the control eye. Further, in swimming tests guided by visual cues, the rats with a BMSC transplant performed significantly better. We believe that BMSC transplantation therapy is effective in treating aged rats with glaucoma. Keywords: glaucoma, stem cell, transplantation, cell therapy, aging

  6. Effect of intravenous transplantation of bone marrow mesenchymal stem cells on neurotransmitters and synapsins in rats with spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    Shaoqiang Chen; Bilian Wu; Jianhua Lin

    2012-01-01

    Bone marrow mesenchymal stem cells were isolated,purified and cultured in vitro by Percoll density gradient centrifugation combined with the cell adherence method.Passages 3-5 bone marrow mesenchymal stem cells were transplanted into rats with traumatic spinal cord injury via the caudal vein.Basso-Beattie-Bresnahan scores indicate that neurological function of experimental rats was significantly improved over transplantation time (1-5 weeks).Expressions of choline acetyltransferase,glutamic acid decarboxylase and synapsins in the damaged spinal cord of rats was significantly increased after transplantation,determined by immunofluorescence staining and laser confocal scanning microscopy.Bone marrow mesenchymal stem cells that had migrated into the damaged area of rats in the experimental group began to express choline acetyltransferase,glutamic acid decarboxylase and synapsins,3 weeks after transplantation.The Basso-Beattie-Bresnahan scores positively correlated with expression of choline acetyltransferase and synapsins.Experimental findings indicate that intravenously transplanted bone marrow mesenchymal stem cells traverse into the damaged spinal cord of rats,promote expression of choline acetyltransferase,glutamic acid decarboxylase and synapsins,and improve nerve function in rats with spinal cord injury.

  7. Mesenchymal stem cells that located in the electromagnetic fields improves rat model of Parkinson's disease

    Directory of Open Access Journals (Sweden)

    Majid Jadidi

    2016-07-01

    Full Text Available Objective(s: The main characteristic of mesenchymal stem cells (MSCs is their ability to produce other cell types. Electromagnetic field (EMF stimulates differentiation of MSCs into other cells. In this study, we investigated whether EMF can effect on the differentiation of MSCs into dopaminergic (DA neurons. Materials and Methods: An EMF with a frequency of 50 Hz and two intensities of 40 and 400 µT 1hr/day was generated around the cells for a week. Afterwards, these cells were injected into the left ventricle of Parkinsonian rats. The rats survived for 2 weeks, and then sampling was performed. Results: The injected cells differentiated into DA neurons and sporadically settled in the substantia nigra pars compacta (SNpc. Transplanted rats exhibited significant partial correction apomorphine-induced rotational behavior compared to Parkinsonian rats (5.0±0.1 vs 7.57±0.08. Results demonstrated that endogenous serum and brain derived neurotrophic factor (BDNF were altered in all experimental groups. The greatest increase was in group of 400 µT EMF in comparison with Parkinsonian rats (398±15 vs. 312±11.79 pg ⁄ mg. Current study have shown that 6-Hydroxydopamine can cause severe loss of dopaminergic neurons (68±6.58, but injected MSCs that exposed to 40 and 400 µT EMF increased dopaminergic neurons in SNpc ( 108±2.33  & 126±3.89 (P

  8. Does injection of metanephric mesenchymal cells improve renal function in rats?

    Directory of Open Access Journals (Sweden)

    Yu-qing Jiao

    2011-01-01

    Full Text Available Chronic kidney disease (CKD is a massive global health-care problem. Cell therapy offers a potential treatment for CKD. The aim of this study was to investigate whether the administration of a population of stem cells could be used to treat adriamycin (ADR-induced glomerulopathy in rats, a form of CKD. We intravenously transplanted metanephric mesenchymal cells (MMCs into rats treated with ADR. We also induced MMC differentiation in vitro using a medium derived from serum and homogenates of ADR-induced glomerulopathy rats. We detected the induction of an early epithelial phenotype (cytokeratin-18 expression and a proximal tubule phenotype (vitamin D receptor expression in vitro, and MMC-derived epithelial cells corresponding to the proximal tubule and glomeruli in vivo. Transplantation of MMCs after induction of glomerulopathy significantly increased the creatinine clearance rate (Ccr, a marker for glomerular filtration rate, but had no significant effect on other parameters (24-hour urinary protein excretion, serum albumin, total cholesterol. In addition, there was no significant difference in blood urea nitrogen or serum creatinine levels in rats with and without ADR administration. Our results indicate that MMCs might survive, engraft and differentiate into renal epithelia in vivo when transplanted into ADR-treated rats. However, further studies are needed to determine whether MMC transplantation improves renal function and causes renal repair in this model.

  9. Mesenchymal Stem Cells Enhance Nerve Regeneration in a Rat Sciatic Nerve Repair and Hindlimb Transplant Model

    Science.gov (United States)

    Cooney, Damon S.; Wimmers, Eric G.; Ibrahim, Zuhaib; Grahammer, Johanna; Christensen, Joani M.; Brat, Gabriel A.; Wu, Lehao W.; Sarhane, Karim A.; Lopez, Joseph; Wallner, Christoph; Furtmüller, Georg J.; Yuan, Nance; Pang, John; Sarkar, Kakali; Lee, W. P. Andrew; Brandacher, Gerald

    2016-01-01

    This study investigates the efficacy of local and intravenous mesenchymal stem cell (MSC) administration to augment neuroregeneration in both a sciatic nerve cut-and-repair and rat hindlimb transplant model. Bone marrow-derived MSCs were harvested and purified from Brown-Norway (BN) rats. Sciatic nerve transections and repairs were performed in three groups of Lewis (LEW) rats: negative controls (n = 4), local MSCs (epineural) injection (n = 4), and systemic MSCs (intravenous) injection (n = 4). Syngeneic (LEW-LEW) (n = 4) and allogeneic (BN-LEW) (n = 4) hindlimb transplants were performed and assessed for neuroregeneration after local or systemic MSC treatment. Rats undergoing sciatic nerve cut-and-repair and treated with either local or systemic injection of MSCs had significant improvement in the speed of recovery of compound muscle action potential amplitudes and axon counts when compared with negative controls. Similarly, rats undergoing allogeneic hindlimb transplants treated with local injection of MSCs exhibited significantly increased axon counts. Similarly, systemic MSC treatment resulted in improved nerve regeneration following allogeneic hindlimb transplants. Systemic administration had a more pronounced effect on electromotor recovery while local injection was more effective at increasing fiber counts, suggesting different targets of action. Local and systemic MSC injections significantly improve the pace and degree of nerve regeneration after nerve injury and hindlimb transplantation. PMID:27510321

  10. Calcium and α-tocopherol suppress cured-meat promotion of chemically induced colon carcinogenesis in rats and reduce associated biomarkers in human volunteers123

    OpenAIRE

    Pierre, Fabrice HF; Martin, Océane CB; Santarelli, Raphaelle L; Taché, Sylviane; Naud, Nathalie; Guéraud, Françoise; Audebert, Marc; Dupuy, Jacques; Meunier, Nathalie; Attaix, Didier; Vendeuvre, Jean-Luc; Mirvish, Sidney S; Kuhnle, Gunter CG; Cano, Noel; Corpet, Denis E

    2013-01-01

    Background: Processed meat intake has been associated with increased colorectal cancer risk. We have shown that cured meat promotes carcinogen-induced preneoplastic lesions and increases specific biomarkers in the colon of rats.

  11. Umbilical cord mesenchymal stem cell transplantation ameliorates burn-induced acute kidney injury in rats.

    Science.gov (United States)

    Lu, Gang; Huang, Sha; Chen, Yongbin; Ma, Kui

    2013-09-01

    Excessive systemic inflammation following burns could lead to acute kidney injury (AKI). Mesenchymal stromal cells (MSCs) suppress immune cell responses and have beneficial effects in various inflammatory-related immune disorders. However, autologous MSCs are not vital enough for the treatment because of the severely burned patients' deleterious condition. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) could be a suitable substitute cell candidate but no data are available on the therapeutic effectiveness of UC-MSCs transplantation for burn injury and its consequences. In this study, UC-MSCs or ulinastatin was administered intravenously in the rats with burn trauma, and the therapeutic effects of UC-MSCs on the survival of severe burn-induced AKI rats and functional protection of kidney were analyzed. Results showed that UC-MSCs promoted the survival and prevented commitment to apoptosis of resident kidney cells and reduced organ microscopic damage in kidneys after thermal trauma. Thus, our study demonstrates that intravenously delivered UC-MSCs protected the host from death caused by kidney injury subsequent to severe burn, identifying UC-MSCs transplantation may be an attractive candidate for cell-based treatments for burns and induced organ damage. PMID:24043673

  12. Effect of dietary supplementation on the prognostic value of urinary and serum 8-isoprostaglandin F2α in chemically-induced mammary carcinogenesis in the rat

    Directory of Open Access Journals (Sweden)

    Białek Sławomir

    2011-03-01

    Full Text Available Abstract Backround The aim of the present study was to assess the effects of zinc or copper and polyphenolic compounds on the 8-isoprostaglandin F2α concentration in the serum and urine of rats with mammary cancer (adenocarcinoma induced with 7,12-dimethylbenz[a]antracene. The research focused on the kinetics of alterations in urinary 8-isoPGF2α at the early stage of carcinogenesis as well as the influence of dietary factors on the process. The impact of selected compounds on the intensity of DMBA - induced carcinogenesis was also assessed. Result and conclusions Administration of DMBA, a compound that inducers mammary tumors in experimental animals, increased the serum and urinary 8-isoPGF2α levels in study rats. In the rat model, diet supplementation with zinc, combined with selected polyphenolic compounds (resveratrol or genistein yielded a statistically significant decrease in the rat serum and urinary biomarker concentration with a simultaneously significant stimulation of carcinogenesis. The results indicate that there is an inverse correlation between the intensity of DMBA-induced carcinogenicity and the level of 8-isoPGF2α in urine and serum of rats.

  13. Early combined treatment with sildenafil and adipose-derived mesenchymal stem cells preserves heart function in rat dilated cardiomyopathy

    Directory of Open Access Journals (Sweden)

    Fu Morgan

    2010-09-01

    Full Text Available Abstract Background We investigated whether early combined autologous adipose-derived mesenchymal stem cell (ADMSC and sildenafil therapy offers an additive benefit in preserving heart function in rat dilated cardiomyopathy (DCM. Methods Adult Lewis rats (n = 8 per group were divided into group 1 (normal control, group 2 (saline-treated DCM rats, group 3 [2.0 × 106 ADMSC implanted into left ventricular (LV myocardium of DCM rats], group 4 (DCM rats with sildenafil 30 mg/kg/day, orally, and group 5 (DCM rats with combined ADMSC-sildenafil. Treatment was started 1 week after DCM induction and the rats were sacrificed on day 90. Results The results showed that mitochondrial protein expressions of connexin43 and cytochrome-C were lowest in group 2, and lower in groups 3 and 4 than in group 5 (p Conclusion Early combined ADMSC/sildenafil is superior to either treatment alone in preserving LV function.

  14. Bone marrow-derived mesenchymal stem cells protect against experimental liver fibrosis in rats

    Institute of Scientific and Technical Information of China (English)

    Dong-Chang Zhao; Jun-Xia Lei; Rui Chen; Wei-Hua Yu; Xiu-Ming Zhang; Shu-Nong Li; Peng Xiang

    2005-01-01

    AIM: Recent reports have shown the capacity of mesenchymal stem cells (MSCs) to differentiate into hepatocytes in vitro and in vivo. MSCs administration could repair injured liver, lung, or heart through reducing inflammation, collagen deposition, and remodeling. These results provide a clue to treatment of liver fibrosis. The aim of this study was to investigate the effect of infusion of bone marrow (BM)-derived MSCs on the experimental liver fibrosis in rats.METHODS: MSCs isolated from BM in male Fischer 344 rats were infused to female Wistar rats induced with carbon tetrachloride (CCl4) or dimethylnitrosamine (DMN).There were two random groups on the 42nd d of CCl4:CCl4/MSCs, to infuse a dose of MSCs alone; CCl4/saline,to infuse the same volume of saline as control. There were another three random groups after exposure to DMN: DMN10/MSCs, to infuse the same dose of MSCs on d 10; DMN10/saline, to infuse the same volume of saline on d 10; DMN20/MSCs, to infuse the same dose of MSCson d 20. The morphological and behavioral changes ofrats were monitored everyday. After 4-6 wk of MSCs administration, all rats were killed and fibrosis index were assessed by histopathology and radioimmunoassay. Smooth muscle alpha-actin (alpha-SMA) of liver were tested by immunohistochemistry and quantified by IBAS 2.5 software. Male rats sex determination region on the Y chromosome (sry) gene were explored by PCR.RESULTS: Compared to controls, infusion of MSCsreduced the mortality rates of incidence in CCl4-induced model (10% vs 20%) and in DMN-induced model (2040% vs 90%).The amount of collagen deposition and alpha-SMA staining was about 40-50% lower in liver of rats with MSCs than that of rats without MSCs. The similar results were observed in fibrosis index. And the effect of the inhibition of fibrogenesis was greater in DMN10/MSCs than in DMN20/MSCs. The sry gene was positive in the liver of rats with MSCs treatment by PCR.CONCLUSION: MSCs treatment can protect against

  15. The Three-Dimensional Collagen Scaffold Improves the Stemness of Rat Bone Marrow Mesenchymal Stem Cells

    Institute of Scientific and Technical Information of China (English)

    Sufang Han; Yannan Zhao; Zhifeng Xiao; Jin Han; Bing Chen; Lei Chen; Jianwu Dai

    2012-01-01

    Mesenchymal stem cells (MSCs) show the great promise for the treatment of a variety of diseases because of their self-renewal and multipotential abilities.MSCs are generally cultured on two-dimensional (2D) substrate in vitro.There are indications that they may simultaneously lose their stemness and multipotentiality as the result of prolonged 2D culture.In this study,we used three-dimensional (3D) collagen scaffolds as rat MSCs carrier and compared the properties of MSCs on 3D collagen scaffolds with monolayer cultured MSCs.The results demonstrated that collagen scaffolds were suitable for rat MSCs adherence and proliferation.More importantly,compared to MSCs under 2D culture,3D MSCs significantly maintained higher expression levels of stemness genes (Oct4,Sox2,Rex-1 and Nanog),yielded high frequencies of colony-forming units-fibroblastic (CFU-F) and showed enhanced osteogenic and adipogenic differentiation efficiency upon induction.Thus,3D collagen scaffolds may be beneficial for expanding rat MSCs while maintaining the stem cell properties in vitro.

  16. The distribution of mesenchymal stem cells after total-body irradiation in rats

    International Nuclear Information System (INIS)

    Objective: To detect the distribution of mesenchymal stem cells(MSCs) after total-body irradiation in rats. Methods: MSCs were cultured and labeled with green fluorescent protein(GFP). Rats were exposed to total-body irradiation(TBI) or TBI plus total brain irradiation, and then MSCs were injected through the tail vein. The Fluorescent MSCs were observed by fluorescence microscope. The MSCs numbers in different organs were determined by quantitative RT-PCR method. Results: GFP-labeled MSCs were obtained. After MSCs were infused to the rats, few of them were observed in the organs of nonirradiated group except for a very low number in the lungs ,bone marrow(BM) and spleen. TBI of 6 Gy increased the engraftment of MSCs in almost all the organs, especially in early response tissues such as the small intestine and BM. TBI of 7 Gy further increased the number of MSCs. The MSCs numbers in the brain and other organs were significantly increased after 20 Gy total brain irradiation in addition to 6 Gy TBI. Conclusions: Radiation injury can induce the aggregation of MSCs. With the increase of radiation dose and severity of radiation injury, a significant increase of MSCs in different organs were observed. Local irradiation can increase the MSCs distribution in the radiation field as well as other organs. (authors)

  17. Therapeutic effect of mesenchymal stem cells transplantation on pulmonary hypertension model rats

    Directory of Open Access Journals (Sweden)

    Ke-yan ZHAO

    2013-11-01

    Full Text Available Objective  To observe the therapeutic effect of mesenchymal stem cells (MSCs transplantation on monocrotaline (MCT-induced pulmonary hypertension in rats. Methods MSCs were isolated from Wistar rats and cultivated by bone marrow adherent culture. The third to fifth passages of MSCs were used for cell transplantation. Forty male Wistar rats were randomly divided into control group, MCT group, MCT/MSCs 5×105 group and MCT/MSCs 1×106 group (10 each. MCT was injected intraperitoneally (60mg/kg, and MSCs were transplanted into rats through external jugular vein. Right ventricular systolic pressure (RVSP of rats was determined, then the animals were sacrificed and the ventricular ratio (i.e. the mass ratio of right to left ventricle plus interventricular septum was calculated. The lung tissue was observed by light microscopy after HE staining, orcein staining and smooth muscle actin immunohistochemical staining. Results Four weeks after MSCs administration, the RVSP and ventricular ratio were 35.6±8.4mmHg and 0.357±0.092 in MCT/MSCs 1×106 group respectively, and significantly lower than those in MCT group (47.2±10.5mmHg and 0.445±0.094, respectively, P0.05. Pathological observation found the intima-media thickness of pulmonary arterioles was 19.2%±3.8% in MCT/MSCs 1×106 group, and significantly thinner than that in MCT group (26.4%±4.9%, P0.05. Conclusion  Intravenous MSCs administration could inhibit the MCT-induced pulmonary hypertension and 1×106 is the optimal number of MSCs for transplantation. DOI: 10.11855/j.issn.0577-7402.2013.10.010

  18. Assessment of the effect of betaine on p16 and c-myc DNA methylation and mRNA expression in a chemical induced rat liver cancer model

    Directory of Open Access Journals (Sweden)

    Ling Wen-hua

    2009-07-01

    Full Text Available Abstract Background The development and progression of liver cancer may involve abnormal changes in DNA methylation, which lead to the activation of certain proto-oncogenes, such as c-myc, as well as the inactivation of certain tumor suppressors, such as p16. Betaine, as an active methyl-donor, maintains normal DNA methylation patterns. However, there are few investigations on the protective effect of betaine in hepatocarcinogenesis. Methods Four groups of rats were given diethylinitrosamine (DEN and fed with AIN-93G diets supplemented with 0, 10, 20 or 40 g betaine/kg (model, 1%, 2%, and 4% betaine, respectively, while the control group, received no DEN, fed with AIN-93G diet. Eight or 15 weeks later, the expression of p16 and c-myc mRNA was examined by Real-time PCR (Q-PCR. The DNA methylation status within the p16 and c-myc promoter was analyzed using methylation-specific PCR. Results Compared with the model group, numbers and areas of glutathione S-transferase placental form (GST-p-positive foci were decreased in the livers of the rats treated with betaine (P . Although the frequency of p16 promoter methylation in livers of the four DEN-fed groups appeared to increase, there is no difference among these groups after 8 or 15 weeks (P > 0.05. Betaine supplementation attenuated the down-regulation of p16 and inhibited the up-regulation of c-myc induced by DEN in a dose-dependent manner (P P . Finally, enhanced antioxidative capacity (T-AOC was observed in both the 2% and 4% betaine groups. Conclusion Our data suggest that betaine attenuates DEN-induced damage in rat liver and reverses DEN-induced changes in mRNA levels.

  19. Assessment of the effect of betaine on p16 and c-myc DNA methylation and mRNA expression in a chemical induced rat liver cancer model

    International Nuclear Information System (INIS)

    The development and progression of liver cancer may involve abnormal changes in DNA methylation, which lead to the activation of certain proto-oncogenes, such as c-myc, as well as the inactivation of certain tumor suppressors, such as p16. Betaine, as an active methyl-donor, maintains normal DNA methylation patterns. However, there are few investigations on the protective effect of betaine in hepatocarcinogenesis. Four groups of rats were given diethylinitrosamine (DEN) and fed with AIN-93G diets supplemented with 0, 10, 20 or 40 g betaine/kg (model, 1%, 2%, and 4% betaine, respectively), while the control group, received no DEN, fed with AIN-93G diet. Eight or 15 weeks later, the expression of p16 and c-myc mRNA was examined by Real-time PCR (Q-PCR). The DNA methylation status within the p16 and c-myc promoter was analyzed using methylation-specific PCR. Compared with the model group, numbers and areas of glutathione S-transferase placental form (GST-p)-positive foci were decreased in the livers of the rats treated with betaine (P < 0.05). Although the frequency of p16 promoter methylation in livers of the four DEN-fed groups appeared to increase, there is no difference among these groups after 8 or 15 weeks (P > 0.05). Betaine supplementation attenuated the down-regulation of p16 and inhibited the up-regulation of c-myc induced by DEN in a dose-dependent manner (P < 0.01). Meanwhile, increases in levels of malondialdehyde (MDA) and glutathione S-transferase (GST) in model, 2% and 4% betaine groups were observed (P < 0.05). Finally, enhanced antioxidative capacity (T-AOC) was observed in both the 2% and 4% betaine groups. Our data suggest that betaine attenuates DEN-induced damage in rat liver and reverses DEN-induced changes in mRNA levels

  20. Genotoxicity of copper oxide nanoparticles with different surface chemistry on rat bone marrow mesenchymal stem cells

    DEFF Research Database (Denmark)

    Zhang, Wenjing; Jiang, Pengfei; Chen, Wei;

    2016-01-01

    The surface chemistry of nanoparticles (NPs) is one of the critical factors determining their cellular responses. In this study, the cytotoxicity and genotoxicity of copper oxide (CuO) NPs with a similar size but different surface chemistry to rat bone marrow mesenchymal stem cells (MSCs) were...... chemistry had influence on the toxicity to some extent too. The intracellular reactive oxygen species (ROS) level of MSCs was then quantified. Finally, the genotoxicity of the CuO NPs was studied by comet assay. The results suggest that the genotoxicity of CuO NPs was mainly dependent on NPs concentration......, and was only slightly influenced by their surface chemistry. The osteogenic and adipogenic differentiation abilities of the MSCs exposed to different CuO NPs were studied by Alizarin Res S and Oil Red O staining. The preliminary results showed that the exposure to 10 μg/mL CuO NPs will not lead to...

  1. Propofol injection combined with bone marrow mesenchymal stem cell transplantation better improves electrophysiological function in the hindlimb of rats with spinal cord injury than monotherapy

    OpenAIRE

    Wang, Yue-Xin; Sun, Jing-Jing; Zhang, Mei; Hou, Xiao-hua; Hong, Jun; Zhou, Ya-Jing; Zhang, Zhi-Yong

    2015-01-01

    The repair effects of bone marrow mesenchymal stem cell transplantation on nervous system damage are not satisfactory. Propofol has been shown to protect against spinal cord injury. Therefore, this study sought to explore the therapeutic effects of their combination on spinal cord injury. Rat models of spinal cord injury were established using the weight drop method. Rats were subjected to bone marrow mesenchymal stem cell transplantation via tail vein injection and/or propofol injection via ...

  2. Propofol combined with bone marrow mesenchymal stem cell transplantation improves electrophysiological function in the hindlimb of rats with spinal cord injury better than monotherapy

    OpenAIRE

    Yue-xin Wang; Jing-jing Sun; Mei Zhang; Xiao-hua Hou; Jun Hong; Ya-jing Zhou; Zhi-yong Zhang

    2015-01-01

    The repair effects of bone marrow mesenchymal stem cell transplantation on nervous system damage are not satisfactory. Propofol has been shown to protect against spinal cord injury. Therefore, this study sought to explore the therapeutic effects of their combination on spinal cord injury. Rat models of spinal cord injury were established using the weight drop method. Rats were subjected to bone marrow mesenchymal stem cell transplantation via tail vein injection and/or propofol injection via ...

  3. In vitro behaviors of rat mesenchymal stem cells on bacterial celluloses with different moduli

    International Nuclear Information System (INIS)

    Compressive moduli of bacteria-synthesized cellulose (BC) were altered by two drying techniques: ambient-air drying and freeze drying. While no significant differences in dry weight were found, their cross-sectional structures and thickness varied greatly. Freeze dried BCs had loose cross-sectional structures and a thickness of ∼ 4.7 mm, whereas air dried BCs had more compacted cross-sectional structures and a thickness of ∼ 0.1 mm. The compressive moduli of the rehydrated freeze dried and rehydrated air dried BCs were measured to be 21.06 ± 0.22 kPa and 90.09 ± 21.07 kPa, respectively. When rat mesenchymal stem cells (rMSCs) were seeded on these BCs, they maintained a round morphology in the first 3 days of cultivation. More spread-out morphology and considerable proliferation on freeze dried BCs were observed in 7 days, but not on air-dried BCs. The cells were further grown for 3 weeks in the absence and presence of differentiation agents. Without using any differentiation agents, no detectable differentiation was noticed for rMSCs further cultivated on both types of BC. With differentiation inducing agents, chondrogenic differentiation, visualized by histological staining, was observed in some area of the rehydrated freeze dried BCs; while osteogenic differentiation was noticed on the stiffer rehydrated air dried BCs. - Graphical abstract: In the presence of induction agents, rat mesenchymal stem cells (rMSCs) preferentially differentiated into osteocytes on stiffer air dried BC films. - Highlights: • Bacterial cellulose (BC) sheets with different moduli generated by drying differently • Air-dried BC exhibited a modulus similar to that of bone. • Freeze-dried BC showed a modulus in the range of that of muscle. • Air-dried BC promoted the differentiation of rMSCs into osteocytes. • Freeze-dried BC promoted the differentiation of rMSCs into chondrocytes

  4. In vitro behaviors of rat mesenchymal stem cells on bacterial celluloses with different moduli

    Energy Technology Data Exchange (ETDEWEB)

    Taokaew, Siriporn [Department of Chemical Engineering, Faculty of Engineering, Chulalongkorn University, Bangkok 10330 (Thailand); Department of Chemical and Biomolecular Engineering, The University of Akron, Akron, OH 44325-3906 (United States); Phisalaphong, Muenduen [Department of Chemical Engineering, Faculty of Engineering, Chulalongkorn University, Bangkok 10330 (Thailand); Zhang Newby, Bi-min, E-mail: bimin@uakron.edu [Department of Chemical and Biomolecular Engineering, The University of Akron, Akron, OH 44325-3906 (United States)

    2014-05-01

    Compressive moduli of bacteria-synthesized cellulose (BC) were altered by two drying techniques: ambient-air drying and freeze drying. While no significant differences in dry weight were found, their cross-sectional structures and thickness varied greatly. Freeze dried BCs had loose cross-sectional structures and a thickness of ∼ 4.7 mm, whereas air dried BCs had more compacted cross-sectional structures and a thickness of ∼ 0.1 mm. The compressive moduli of the rehydrated freeze dried and rehydrated air dried BCs were measured to be 21.06 ± 0.22 kPa and 90.09 ± 21.07 kPa, respectively. When rat mesenchymal stem cells (rMSCs) were seeded on these BCs, they maintained a round morphology in the first 3 days of cultivation. More spread-out morphology and considerable proliferation on freeze dried BCs were observed in 7 days, but not on air-dried BCs. The cells were further grown for 3 weeks in the absence and presence of differentiation agents. Without using any differentiation agents, no detectable differentiation was noticed for rMSCs further cultivated on both types of BC. With differentiation inducing agents, chondrogenic differentiation, visualized by histological staining, was observed in some area of the rehydrated freeze dried BCs; while osteogenic differentiation was noticed on the stiffer rehydrated air dried BCs. - Graphical abstract: In the presence of induction agents, rat mesenchymal stem cells (rMSCs) preferentially differentiated into osteocytes on stiffer air dried BC films. - Highlights: • Bacterial cellulose (BC) sheets with different moduli generated by drying differently • Air-dried BC exhibited a modulus similar to that of bone. • Freeze-dried BC showed a modulus in the range of that of muscle. • Air-dried BC promoted the differentiation of rMSCs into osteocytes. • Freeze-dried BC promoted the differentiation of rMSCs into chondrocytes.

  5. Bone Marrow Mesenchymal Stem Cell and Vein Conduit on Sciatic Nerve Repair in Rats

    Science.gov (United States)

    Seyed Foroutan, Kamal; Khodarahmi, Ali; Alavi, Hootan; Pedram, Sepehr; Baghaban Eslaminejad, Mohamad Reza; Bordbar, Sima

    2015-01-01

    Background: Peripheral nerve repair with sufficient functional recovery is an important issue in reconstructive surgery. Stem cells have attracted extensive research interest in recent years. Objectives: The purpose of this study was to compare the vein conduit technique, with and without the addition of mesenchymal stem cells in gap-less nerve injury repair in rats. Materials and Methods: In this study, 36 Wistar rats were randomly allocated to three groups: In the first group, nerve repair was performed with simple neurorrhaphy (control group), in the second group, nerve repair was done with vein conduit over site (vein conduit group) and in the third group, bone marrow stem cells were instilled into the vein conduit (stem cell group) after nerve repair with vein conduit over site. Six weeks after the intervention, the sciatic function index, electrophysiological study and histological examination were performed. Results: All animals tolerated the surgical procedures and survived well. The sciatic function index and latency were significantly improved in the vein conduit (P = 0.04 and 0.03, respectively) and stem cell group (P = 0.02 and 0.03, respectively) compared with the control group. No significant difference was observed in sciatic function and latency between the vein conduit and stem-cell groups. Moreover, histological analysis showed no significant difference in regenerative density between these two groups. Conclusions: The results of this study showed that the meticulous microsurgical nerve repair, which was performed using the vein tubulization induced significantly better sciatic nerve regeneration. However, the addition of bone marrow mesenchymal stem cell to vein conduit failed to promote any significant changes in regeneration outcome. PMID:25825699

  6. The phytoestrogenic Cyclopia extract, SM6Met, increases median tumor free survival and reduces tumor mass and volume in chemically induced rat mammary gland carcinogenesis.

    Science.gov (United States)

    Visser, Koch; Zierau, Oliver; Macejová, Dana; Goerl, Florian; Muders, Michael; Baretton, Gustavo B; Vollmer, Günter; Louw, Ann

    2016-10-01

    SM6Met, a phytoestrogenic extract of Cyclopia subternata indigenous to the Western Cape province of South Africa, displays estrogenic attributes with potential for breast cancer chemoprevention. In this study, we report that SM6Met, in the presence of estradiol, induces a significant cell cycle G0/G1 phase arrest similar to the selective estrogen receptor modulator, tamoxifen. Furthermore, as a proof of concept, in the N-Methyl-N-nitrosourea induced rat mammary gland carcinogenesis model, SM6Met increases tumor latency by 7days and median tumor free survival by 42 days, while decreasing palpable tumor frequency by 32%, tumor mass by 40%, and tumor volume by 53%. Therefore, the current study provides proof of concept that SM6Met has definite potential as a chemopreventative agent against the development and progression of breast cancer. PMID:27142456

  7. Human umbilical cord blood-derived mesenchymal stem cells promote regeneration of crush-injured rat sciatic nerves

    Institute of Scientific and Technical Information of China (English)

    Mi-Ae Sung; Jong-Ho Lee; Hun Jong Jung; Jung-Woo Lee; Jin-Yong Lee; Kang-Mi Pang; Sang Bae Yoo; Mohammad S. Alrashdan; Soung-Min Kim; Jeong Won Jahng

    2012-01-01

    Several studies have demonstrated that human umbilical cord blood-derived mesenchymal stem cells can promote neural regeneration following brain injury. However, the therapeutic effects of human umbilical cord blood-derived mesenchymal stem cells in guiding peripheral nerve regeneration remain poorly understood. This study was designed to investigate the effects of human umbilical cord blood-derived mesenchymal stem cells on neural regeneration using a rat sciatic nerve crush injury model. Human umbilical cord blood-derived mesenchymal stem cells (1 × 106) or a PBS control were injected into the crush-injured segment of the sciatic nerve. Four weeks after cell injection, brain-derived neurotrophic factor and tyrosine kinase receptor B mRNA expression at the lesion site was increased in comparison to control. Furthermore, sciatic function index, Fluoro Gold-labeled neuron counts and axon density were also significantly increased when compared with control. Our results indicate that human umbilical cord blood-derived mesenchymal stem cells promote the functional recovery of crush-injured sciatic nerves.

  8. Mesenchymal stem cells protect against the tissue fibrosis of ketamine-induced cystitis in rat bladder.

    Science.gov (United States)

    Kim, Aram; Yu, Hwan Yeul; Heo, Jinbeom; Song, Miho; Shin, Jung-Hyun; Lim, Jisun; Yoon, Soo-Jung; Kim, YongHwan; Lee, Seungun; Kim, Seong Who; Oh, Wonil; Choi, Soo Jin; Shin, Dong-Myung; Choo, Myung-Soo

    2016-01-01

    Abuse of the hallucinogenic drug ketamine promotes the development of lower urinary tract symptoms that resemble interstitial cystitis. The pathophysiology of ketamine-induced cystitis (KC) is largely unknown and effective therapies are lacking. Here, using a KC rat model, we show the therapeutic effects of human umbilical cord-blood (UCB)-derived mesenchymal stem cells (MSCs). Daily injection of ketamine to Sprague-Dawley rats for 2-weeks resulted in defective bladder function, indicated by irregular voiding frequency, increased maximum contraction pressure, and decreased intercontraction intervals and bladder capacity. KC bladders were characterized by severe mast-cell infiltration, tissue fibrosis, apoptosis, upregulation of transforming growth factor-β signaling related genes, and phosphorylation of Smad2 and Smad3 proteins. A single administration of MSCs (1 × 10(6)) into bladder tissue not only significantly ameliorated the aforementioned bladder voiding parameters, but also reversed the characteristic histological and gene-expression alterations of KC bladder. Treatment with the antifibrotic compound N-acetylcysteine also alleviated the symptoms and pathological characteristics of KC bladder, indicating that the antifibrotic capacity of MSC therapy underlies its benefits. Thus, this study for the first-time shows that MSC therapy might help to cure KC by protecting against tissue fibrosis in a KC animal model and provides a foundation for clinical trials of MSC therapy. PMID:27481042

  9. Umbilical cord-derived mesenchymal stem cells alleviate liver fibrosis in rats

    Science.gov (United States)

    Chai, Ning-Li; Zhang, Xiao-Bin; Chen, Si-Wen; Fan, Ke-Xing; Linghu, En-Qiang

    2016-01-01

    AIM: To evaluate the efficacy of umbilical cord-derived mesenchymal stem cells (UC-MSCs) transplantation in the treatment of liver fibrosis. METHODS: Cultured human UC-MSCs were isolated and transfused into rats with liver fibrosis induced by dimethylnitrosamine (DMN). The effects of UC-MSCs transfusion on liver fibrosis were then evaluated by histopathology; serum interleukin (IL)-4 and IL-10 levels were also measured. Furthermore, Kupffer cells (KCs) in fibrotic livers were isolated and cultured to analyze their phenotype. Moreover, UC-MSCs were co-cultured with KCs in vitro to assess the effects of UC-MSCs on KCs’ phenotype, and IL-4 and IL-10 levels were measured in cell culture supernatants. Finally, UC-MSCs and KCs were cultured in the presence of IL-4 antibodies to block the effects of this cytokine, followed by phenotypical analysis of KCs. RESULTS: UC-MSCs transfused into rats were recruited by the injured liver and alleviated liver fibrosis, increasing serum IL-4 and IL-10 levels. Interestingly, UC-MSCs promoted mobilization of KCs not only in fibrotic livers, but also in vitro. Co-culture of UC-MSCs with KCs resulted in increased production of IL-4 and IL-10. The addition of IL-4 antibodies into the co-culture system resulted in decreased KC mobilization. CONCLUSION: UC-MSCs could increase IL-4 and promote mobilization of KCs both in vitro and in vivo, subsequently alleviating the liver fibrosis induced by DMN.

  10. Cysteine: A Novel Neural Inducer for Rat Bone Marrow Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Malek Soleimani Mehranjani

    2014-06-01

    Full Text Available Objective: Mesenchymal stem cells (MSCs can differentiate into various cell types. Since cysteine has structural similarities to neuronal inducers β-mercaptoethanol and glutathione, we examined its effect on neural induction of rat bone marrow MSCs. Materials and Methods: In this experimental study, cells were treated in a medium containing 1mM cysteine for 24 hours prior to treatment with neuron inducing medium containing 10 mM cysteine for 1, 2 and 3 hours. Cell viability and morphology were assessed by 3-(4,5-dimethylthiazol-2-Yl-2,5-diphenyltetrazolium bromide (MTT assay and, Hoechst, propidium iodide and acridine orange staining respectively. Expression of nestin and β-Tubulin III genes, as neural cell-specific markers, was studied reverse transcription polymerase chain reaction (RT-PCR. The data was statistically analyzed using One-Way ANOVA and Tukey’s test and p<0.05 was considered significant. Results: After 3 hours of treatment, neuron like morphology with a considerable expression of nestin and β-Tubulin III genes was apparent. The mean cell viability was not significantly different at 1, 2 and 3 hours following induction, compared with the control cells. Conclusion: Cysteine can induce neural features in rat bone marrow MSCs without reducing cell viability. Therefore, it can be considered as a safer alternative to toxic neural inducer agents such as β-mercaptoethanol.

  11. Magnetic labeling and in vitro MR imaging of rat bone marrow mesenchymal stem cells

    International Nuclear Information System (INIS)

    Objective: To label rat bone marrow mesenchymal stem cells with feridex combined with poly-l-lysine (PLL), and to determine the feasibility of detection of magnetically labeled stem cells with MR imaging. Methods: Feridex were incubated with PLL for 1 hour to obtain a complex of feridex-PLL. Mesenchymal stem cells isolated from the bone marrows of Wistar rats were cultured and expanded. By the 4th passage, cells were co-incubated overnight with the feridex-PLL complex. Prussian blue staining for demonstrating intracytoplastic nanoparticles and trypan-blue exclusion test for cell viability were performed respectively at 24 h, 1 w, 2 w, 3 w after labeling. MR imaging of cell suspensions was performed by using T1WI, T2WI and T2* WI sequences at a clinical 1.5 T MR system. Results: Numerous intracytoplastic iron particles were stained with Prussian blue. With division of stern cells, the stained particles were seen decreased gradually. Trypan blue exclusion test at 24 h, 1 w, 2 w and 3 w showed that the viability of the labeled cells was 91.00%, 93.00%, 91.75%, and 92.50%, not significantly different with that of nonlabeled cells (P>0.05). For 103, 104 and l05 cells, T2 signal intensity decreased by 63.75%, 82.31% and 91.92% respectively, T2* signal intensity decreased by 68.24%, 83.01%, and 93.94% respectively. For 105 labeled cells, T2* signal intensity decreased by 93.75%, 75.92%, 41.75% and 8.83 % respectively at 24 h, 1 w, 2 w and 3 w after labeling. Conclusion: Magnetic labeling of rat bone marrow stem cells with feridex-PLL complex is feasible, efficient and safe. T2* WI is the most sensitive sequence to detect the labeled cells. The degree of T2 signal decreasing may be related to the cell count and division phase. (authors)

  12. The osteogenic differentiation stimulating activity of Sea cucumber methanolic crude extraction on rat bone marrow mesenchymal stem cells

    OpenAIRE

    Javad Baharara; Elaheh Amini; Mohammad Amin Kerachian; Mozhgan Soltani

    2014-01-01

    Objective(s): Sea cucumber derived bioactive compound is considered efficient in treatment of bone disorders. This study was performed to evaluate the effect of this extract on differentiation of rat bone marrow mesenchymal stem cells (rBMMSc) into osteogenic lineage. Materials and Methods: Isolated rBMMSc were grown in DMEM supplemented with 10% FBS. The cells were exposed to different concentration of extract. After 21 days, Alizarin red staining, alkaline phosphatase assay and RT-PCR were ...

  13. Effects of immunosuppressants, FK506 and cyclosporin A, on the osteogenic differentiation of rat mesenchymal stem cells

    OpenAIRE

    Byun, Yu-Kyung; Kim, Kyoung-Hwa; Kim, Su-Hwan; Kim, Young-Sung; Koo, Ki-Tae; Kim, Tai-Il; Seol, Yang-Jo; Ku, Young; Rhyu, In-Chul; Lee, Yong-Moo

    2012-01-01

    Purpose The purpose of this study was to investigate the effects of the immunosuppressants FK506 and cyclosporin A (CsA) on the osteogenic differentiation of rat mesenchymal stem cells (MSCs). Methods The effect of FK506 and CsA on rat MSCs was assessed in vitro. The MTT assay was used to determine the deleterious effect of immunosuppressants on stem cell proliferation at 1, 3, and 7 days. Alkaline phosphatase (ALP) activity was analyzed on days 3, 7, and 14. Alizarin red S staining was done ...

  14. Do adipose tissue-derived mesenchymal stem cells ameliorate Parkinson's disease in rat model?

    Science.gov (United States)

    Ahmed, Hh; Salem, Am; Atta, Hm; Ghazy, Ma; Aglan, Ha

    2014-12-01

    Parkinson's disease (PD) is a common neurodegenerative disorder in middle-aged and elderly people. This study aimed to elucidate the role of mesenchymal stem cells (MSCs) in management of PD in ovariectomized rat model. MSCs were excised from adipose tissue of both the omentum and the inguinal fat pad of male rats, grown, and propagated in culture; then characterized morphologically; and by the detection of surface markers gene expression. In this study, 40 ovariectomized animals were classified into 5 groups; group 1 was ovariectomized control, groups 2 to 5 were subcutaneously administered with rotenone for 14 days after 1 month of ovariectomy for induction of PD. Group 2 was left untreated; groups 3, 4, and 5 were treated with Sinemet(®), Cerebrolysin(®), and a single dose of adipose tissue-derived MSCs (ADMSCs), respectively. Y-chromosome gene (sry) was assessed by polymerase chain reaction (PCR) in brain tissue of the female rats. Serum transforming growth factor β (TGF-β), monocyte chemoattractant protein 1 (MCP-1), and brain-derived neurotrophic factor (BDNF) levels were assayed using enzyme-linked immunosorbent assay technique. Brain dopamine level was assayed fluorometrically, while brain tyrosine hydroxylase (TH) gene expression was detected by semiquantitative real-time PCR. The PD group showed significant increase in serum TGF-β and MCP-1 levels associated with significant decrease in serum BDNF, brain dopamine, and brain TH gene expression levels. In contrast, all treatments produce significant decrease in serum TGF-β and MCP-1 levels in concomitant with significant increase in serum BDNF, brain dopamine, and brain TH gene expression levels. In conclusion, the observed improvements in the studied biomarkers due to ADMSCs infusion might be attributed to their immunomodulatory, anti-inflammatory, and neurotrophic effects. PMID:24567299

  15. Transplantation of human amniotic mesenchymal stem cells promotes neurological recovery in an intracerebral hemorrhage rat model.

    Science.gov (United States)

    Zhou, Honglong; Zhang, Hongri; Yan, Zhongjie; Xu, Ruxiang

    2016-06-24

    Human amniotic membrane mesenchymal stem cells (hAMSCs) have recently been suggested as ideal candidate stem cells for cell-based therapy. Many studies have reported the therapeutic effects of hAMSCs in numerous disease models. However, no studies have used hAMSCs to treat intracerebral hemorrhage (ICH). In the present study, we examined the therapeutic potential of hAMSCs in a rat model of ICH, and characterized the possible mechanisms of action. Adult male Wistar rats were subjected to ICH by intrastriatal injection of VII collagenase, and then were intracerebrally administered hAMSCs, fibroblasts, or phosphate-buffered saline (PBS) at 24 h after ICH. Compared with the fibroblasts and the PBS control, hAMSCs treatment significantly promoted neurological recovery, and reduced the numbers of ED1(+) activated microglia, as well as myeloperoxidase (MPO(+)), and caspase-3(+) cells in the brain injury model. In addition, hAMSCs treatment significantly increased the expression of brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF) in the injured brain, and promoted neurogenesis and angiogenesis, compared with the fibroblasts and the PBS control. The transplanted hAMSCs survived for at least 27 days and were negative for β-tubulin III and glial fibrillary acidic protein (GFAP). Taken together, the results suggest that hAMSCs treatment significantly promotes neurological recovery in rats after ICH. The mechanism of action could be mediated by inhibition of inflammation and apoptosis, increasing neurotrophic factor expression, and promotion of neurogenesis and angiogenesis. Thus, hAMSCs are candidate stem cells for the treatment of ICH. PMID:27188654

  16. In vivo MR tracking of magnetically labeled mesenchymal stem cells in rat liver after intrasplenic transplantation

    International Nuclear Information System (INIS)

    Objective: To evaluate the 1.5-T magnetic resonance imaging system to depict and track in vivo of magnetically labeled bone mesenchymal stem cells (BMSCs) in rat liver after intrasplenic transplantation. Methods: Rat BMSCs were isolated, purified, expanded and then incubated with home synthesized Fe2O3-PLL. Prussian blue stain was performed for showing intracellular irons. Acute liver damage was induced by subcutaneous injection of carbon tetrachloride in 12 recipient rats. The cells allotransplantation was performed by intrasplenic injection with magnetically labeled (experimental group, n=6) or unlabeled BMSCs (control group, n=6). Serial MRI were performed before and 3 hours, 3, 7, 14 days after transplantation. Signal-to-noise ratio (SNR) of liver on T2*-weighted MR imaging obtained before and after injection were measured and compared. MR imaging findings were compared histologically with histology. Results: Rats BMSCs could be effectively labeled and the labeling efficiency was almost 100%. Iron-containing intracytoplasmatic vesicles could be observed clearly with prussian blue staining. SNR of rat livers in experimental group and control group before and 3 hours, 3, 7, 14 days after transplantation were 19.53±2.30, 3.28±1.06, 7.34±2.10, 10.25±3.96, 15.50±3.73; 20.20±4.35, 21.20±4.43, 19.13±2.80, 21.43±5.45, 19.07±4.80, respectively. SNR decreased significantly in the experimental group liver 3 hours, 3, 7 days after injection of BMSCs (Dunnett test, P0.05). In control group, SNR demonstrated no significant differences among different time points (ANOVA, P>0.05). Results of histological analysis confirmed homing of labeled BMSCs in liver, primarily distributing in areas around centrolobular vein. Conclusion: The BMSCs can be effectively labeled with Fe2O3-PLL. 1.5-T MR imaging can monitor in vivo of magnetically labeled BMSCs in liver after intrasplenic transplantation. It potentially opens a new area of investigation for delivering stem cells

  17. Chinese preparation Xuesaitong promotes the mobilization of bone marrow mesenchymal stem cells in rats with cerebral infarction.

    Science.gov (United States)

    Zhang, Jin-Sheng; Zhang, Bao-Xia; Du, Mei-Mei; Wang, Xiao-Ya; Li, Wei

    2016-02-01

    After cerebral ischemia, bone marrow mesenchymal stem cells are mobilized and travel from the bone marrow through peripheral circulation to the focal point of ischemia to initiate tissue regeneration. However, the number of bone marrow mesenchymal stem cells mobilized into peripheral circulation is not enough to exert therapeutic effects, and the method by which blood circulation is promoted to remove blood stasis influences stem cell homing. The main ingredient of Xuesaitong capsules is Panax notoginseng saponins, and Xuesaitong is one of the main drugs used for promoting blood circulation and removing blood stasis. We established rat models of cerebral infarction by occlusion of the middle cerebral artery and then intragastrically administered Xuesaitong capsules (20, 40 and 60 mg/kg per day) for 28 successive days. Enzyme-linked immunosorbent assay showed that in rats with cerebral infarction, middle- and high-dose Xuesaitong significantly increased the level of stem cell factors and the number of CD117-positive cells in plasma and bone marrow and significantly decreased the number of CD54- and CD106-positive cells in plasma and bone marrow. The effect of low-dose Xuesaitong on these factors was not obvious. These findings demonstrate that middle- and high-dose Xuesaitong and hence Panax notoginseng saponins promote and increase the level and mobilization of bone marrow mesenchymal stem cells in peripheral blood. PMID:27073383

  18. A comparative study on the transplantation of different concentrations of human umbilical mesenchymal cells into diabetic rats

    Institute of Scientific and Technical Information of China (English)

    Jia-Hui; Kong; Dan; Zheng; Song; Chen; Hong-Tao; Duan; Yue-Xin; Wang; Meng; Dong; Jian; Song

    2015-01-01

    AIM: To observe the effects of intravitreal injections of different concentrations of human umbilical mesenchymal stem cells on retinopathy in rats with diabetes mellitus.METHODS: Healthy and adult male Sprague-Dawley(SD) rats were randomly assigned to a normal control group(group A), a diabetic retinopathy(DR) blank control group(group B), a high-concentration transplantation group(group C), a low-concentration transplantation group(group D) and a placebo transplantation group(group E). The expression of nerve growth factor(NGF)protein in the retinal layers was detected by immunohistochemical staining at 2, 4, 6 and 8wk.RESULTS: The expression of NGF was positive in group A and most positive in the retinal ganglion cell layer. In groups B and E, the expression of NGF was positive 2wk after transplantation and showed an increase in all layers. However, the level of expression had decreased in all layers at 4wk and was significantly reduced at 8wk. In groups C and D, the expression of NGF had increased at 2wk and continued to increase up to 8wk. The level of expression in group C was much higher than that in group D.CONCLUSION: DR can be improved by intravitreal injection of human umbilical mesenchymal stem cells.High concentrations of human umbilical mesenchymal stem cells confer a better protective effect on DR than low concentrations.

  19. Curcumin increases rat mesenchymal stem cell osteoblast differentiation but inhibits adipocyte differentiation

    Directory of Open Access Journals (Sweden)

    Qiaoli Gu

    2012-01-01

    Full Text Available Background: Curcumin is a phenolic natural product isolated from the rhizome of Curcuma longa (turmeric and has effects on bone health and fat formation. The bone marrow mesenchymal stem cells (MSCs are multipotent cells capable of differentiating into osteoblasts and adipocytes. Osteoblast differentiation of MSCs can be a result of upregulation of heme oxygenase (HO-1 expression. Curcumin can potently induce HO-1 expression. Objective: The present study describes the effects of curcumin on rat MSC (rMSCs differentiation into osteoblasts and adipocytes. Materials and Methods: Rat bone marrow MSCs were isolated and treated with or without curcumin. Osteoblast differentiation was confirmed and determined by alkaline phosphatase (ALP activity, mineralized nodule formation, the expression of Runx2 (runt-related transcription factor 2 and osteocalcin. Adipocyte differentiation was determined by Oil red O staining and the expression of peroxisome proliferator-activated receptor-γ 2 (PPARγ2 and CCAAT/enhancer-binding protein (C/EBP α. Results: Curcumin increased ALP activity and osteoblast-specific mRNA expression of Runx2 and osteocalcin when rMSCs were cultured in osteogenic medium. In contrast, curcumin decreased adipocyte differentiation and inhibited adipocyte-specific mRNA expression of PPARγ2 and C/EBPα when rMSCs were cultured in adipogenic medium. HO-1 expression was increased during osteogenic differentiation of rMSCs. Conclusions: These findings demonstrate that curcumin can promote osteogenic differentiation of rMSCs and inhibit adipocyte formation. The effect of curcumin on osteogenic differentiation of rMSCs is correlated with HO-1 expression.

  20. The effect of rat bone marrow derived mesenchymal stem cells transplantation for restoration of olfactory disorder.

    Science.gov (United States)

    Jo, Hyogyeong; Jung, Minyoung; Seo, Dong Jin; Park, Dong Joon

    2015-11-13

    The purpose of the study was to investigate the effect of bone marrow-derived mesenchymal stem cells (BMSCs) transplantation on olfactory epithelium (OE) of morphologic and functional restoration following neural Sensorineural Disorder in rats. Except the Normal group, twenty-one rats underwent Triton X-100 (TX-100) irrigation to induce degeneration of OE, and then BMSCs and PBS were treated from the both medial canthus to the rear part of the both nasal cavity into the experimental group and then were observed for restoration according to time point. At two and four weeks after transplantation with BMSCs, restoration of OE was observed with olfactory marker protein (OMP) and behavioral test. And we observed the expression of OMP, nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF). After TX-100 irrigation, the OE almost disappeared in 3 days. At four weeks after transplantation with BMSCs, the thickness and cellular composition of OE was considerably restored to normal group and expression of OMP was markedly increased when compared with PBS group and reduced the searching time in the behavioral test. Furthermore at two weeks after treatment with BMSCs, expression of NGF and BDNF was greatly increased when compared with PBS group. However at four weeks after treatment with BMSCs, expression of NGF and BDNF was slightly decreased. Our results suggest the BMSCs transplantation affect restoration of OE and olfaction, most likely via regulation of the neurotrophic factor expression, especially the expression of NGF and BDNF and has a possibility of a new therapeutic strategy for the treatment of olfactory disorder caused by the degeneration of OE. PMID:26427869

  1. Effects of strontium on proliferation and differentiation of rat bone marrow mesenchymal stem cells

    International Nuclear Information System (INIS)

    Highlights: ► Strontium ranelate (SrR) inhibits proliferation of BMMSCs. ► SrR increases osteoblastic but decreases adipocytic differentiation of BMMSCs. ► SrR increases expression of Runx2, BSP and OCN by BMMSCs in osteogenic medium. ► SrR decreases expression of PPARγ, aP2/ALBP and LPL by BMMSCs in adipogenic medium. -- Abstract: Strontium ranelate (SrR) was an effective anti-osteoporotic drug to increase bone formation and decrease bone resorption. However, reports about the effect of SR on osteoblastic and adipocytic differentiation from bone marrow mesenchymal stem cells (BMMSCs) are limited. The purpose of this study is to evaluate whether SrR affects the ability of BMMSCs to differentiate into osteoblasts or adipocytes. Rat BMMSCs were identified by flow cytometry and exposed to SR (0.1 and 1.0 mM Sr2+) under osteogenic or adipogenic medium for 1 and 2 weeks. The proliferation and differentiation of BMMSCs were analyzed by MTT, alkaline phosphatase (ALP), Oil red O staining, quantitative real-time RT-PCR and Western blot assays. SrR significantly inhibited the proliferation, increased osteoblastic but decreased adipocytic differentiation of rat BMMSCs dose-dependently. In osteogenic medium, SrR increased the expression of ALP, the mRNA levels of Cbfa1/Runx2, bone sialoprotein, and osteocalcin by RT-PCR, and the protein levels of Cbfa1/Runx2 by Western blot. In adipogenic medium, SrR decreased the mRNA levels of PPARγ2, adipocyte lipid-binding protein 2 (aP2/ALBP), and lipoprotein lipase (LPL) by RT-PCR, and the protein expression of PPARγ in Western blot analysis. These results indicated that the effects of SrR to promote osteoblastic but inhibit adipocytic differentiation of BMMSCs might contribute to its effect on osteoporosis treatment.

  2. Synergistic Effects of Electroacupuncture and Mesenchymal Stem Cells on Intestinal Ischemia/Reperfusion Injury in Rats.

    Science.gov (United States)

    Geng, Yanxia; Chen, Dong; Zhou, Jiang; Lu, Jun; Chen, Mingqi; Zhang, Haidong; Wang, Xing

    2016-08-01

    Electroacupuncture (EA) and transplantation of bone marrow mesenchymal stem cells (MSCs) are both promising therapeutic applications for intestinal disorders. The current study examined their combined effect on rat intestinal ischemia/reperfusion (I/R) injury and the possible mechanism. Five groups were performed: con group (shame operation),I/R group (model group), MSC group (I/R + MSC), EA group (I/R + EA), and combined group (I/R + MSC + EA). Intestinal histological damage, crypt cell proliferation degree, mucosal cytokines expression, and levels of inflammation factors were studied for each group. Compared with the I/R group, crypt cell proliferation index and mucosal mRNA concentration of SDF-1, CXCR4, EGF, EGFR in MSC group and EA group were significantly increased, with mucosal NF-кBp65 and serum inflammation factor (TNF-α, IL-6) levels significantly decreased. Above all of these indicators except NF-кBp65 were improved more notably in combined group than the other two treatment groups. Chiu's score was only ameliorated remarkably in the combined group. The combined treatment of MSC transplantion and electroacupuncture could protect intestinal mucosal barrier from I/R injury. PMID:27221138

  3. Bone marrow mesenchymal stem cells overexpressing human basic fibroblast growth factor increase vasculogenesis in ischemic rats

    Directory of Open Access Journals (Sweden)

    J.C. Zhang

    2014-10-01

    Full Text Available Administration or expression of growth factors, as well as implantation of autologous bone marrow cells, promote in vivo angiogenesis. This study investigated the angiogenic potential of combining both approaches through the allogenic transplantation of bone marrow-derived mesenchymal stem cells (MSCs expressing human basic fibroblast growth factor (hbFGF. After establishing a hind limb ischemia model in Sprague Dawley rats, the animals were randomly divided into four treatment groups: MSCs expressing green fluorescent protein (GFP-MSC, MSCs expressing hbFGF (hbFGF-MSC, MSC controls, and phosphate-buffered saline (PBS controls. After 2 weeks, MSC survival and differentiation, hbFGF and vascular endothelial growth factor (VEGF expression, and microvessel density of ischemic muscles were determined. Stable hbFGF expression was observed in the hbFGF-MSC group after 2 weeks. More hbFGF-MSCs than GFP-MSCs survived and differentiated into vascular endothelial cells (P<0.001; however, their differentiation rates were similar. Moreover, allogenic transplantation of hbFGF-MSCs increased VEGF expression (P=0.008 and microvessel density (P<0.001. Transplantation of hbFGF-expressing MSCs promoted angiogenesis in an in vivo hind limb ischemia model by increasing the survival of transplanted cells that subsequently differentiated into vascular endothelial cells. This study showed the therapeutic potential of combining cell-based therapy with gene therapy to treat ischemic disease.

  4. Bone marrow mesenchymal stem cells overexpressing human basic fibroblast growth factor increase vasculogenesis in ischemic rats

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    Zhang, J.C. [Department of Vascular Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China); Zheng, G.F. [Department of Vascular Surgery, The People' s Hospital of Ganzhou, Ganzhou (China); Wu, L.; Ou Yang, L.Y.; Li, W.X. [Department of Vascular Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China)

    2014-08-08

    Administration or expression of growth factors, as well as implantation of autologous bone marrow cells, promote in vivo angiogenesis. This study investigated the angiogenic potential of combining both approaches through the allogenic transplantation of bone marrow-derived mesenchymal stem cells (MSCs) expressing human basic fibroblast growth factor (hbFGF). After establishing a hind limb ischemia model in Sprague Dawley rats, the animals were randomly divided into four treatment groups: MSCs expressing green fluorescent protein (GFP-MSC), MSCs expressing hbFGF (hbFGF-MSC), MSC controls, and phosphate-buffered saline (PBS) controls. After 2 weeks, MSC survival and differentiation, hbFGF and vascular endothelial growth factor (VEGF) expression, and microvessel density of ischemic muscles were determined. Stable hbFGF expression was observed in the hbFGF-MSC group after 2 weeks. More hbFGF-MSCs than GFP-MSCs survived and differentiated into vascular endothelial cells (P<0.001); however, their differentiation rates were similar. Moreover, allogenic transplantation of hbFGF-MSCs increased VEGF expression (P=0.008) and microvessel density (P<0.001). Transplantation of hbFGF-expressing MSCs promoted angiogenesis in an in vivo hind limb ischemia model by increasing the survival of transplanted cells that subsequently differentiated into vascular endothelial cells. This study showed the therapeutic potential of combining cell-based therapy with gene therapy to treat ischemic disease.

  5. Bone marrow mesenchymal stem cells overexpressing human basic fibroblast growth factor increase vasculogenesis in ischemic rats

    International Nuclear Information System (INIS)

    Administration or expression of growth factors, as well as implantation of autologous bone marrow cells, promote in vivo angiogenesis. This study investigated the angiogenic potential of combining both approaches through the allogenic transplantation of bone marrow-derived mesenchymal stem cells (MSCs) expressing human basic fibroblast growth factor (hbFGF). After establishing a hind limb ischemia model in Sprague Dawley rats, the animals were randomly divided into four treatment groups: MSCs expressing green fluorescent protein (GFP-MSC), MSCs expressing hbFGF (hbFGF-MSC), MSC controls, and phosphate-buffered saline (PBS) controls. After 2 weeks, MSC survival and differentiation, hbFGF and vascular endothelial growth factor (VEGF) expression, and microvessel density of ischemic muscles were determined. Stable hbFGF expression was observed in the hbFGF-MSC group after 2 weeks. More hbFGF-MSCs than GFP-MSCs survived and differentiated into vascular endothelial cells (P<0.001); however, their differentiation rates were similar. Moreover, allogenic transplantation of hbFGF-MSCs increased VEGF expression (P=0.008) and microvessel density (P<0.001). Transplantation of hbFGF-expressing MSCs promoted angiogenesis in an in vivo hind limb ischemia model by increasing the survival of transplanted cells that subsequently differentiated into vascular endothelial cells. This study showed the therapeutic potential of combining cell-based therapy with gene therapy to treat ischemic disease

  6. Protective effect of bone marrow mesenchymal stem cells combined with erythropoietin therapy on spinal cord injury rat model

    Institute of Scientific and Technical Information of China (English)

    Peng Xie; Wen-Hui Ruan

    2016-01-01

    Objective:To study the protective effect of bone marrow mesenchymal stem cells combined with erythropoietin therapy on spinal cord injury rat model.Methods: SD rats were selected as experimental animals, spinal cord injury rat model was built by striking spinal cord with Hatteras Instruments PCI3000, and model rats were divided into control group, bone marrow mesenchymal stem cells (BMSCs) group, erythropoietin (EPO) group and BMSCs combined with EPO group according to different treatment methods. Then number of apoptotic cells in spinal cord tissue, contents of neural markers and neurotrophic factors as well as expression of apoptosis and injury molecules was detected.Results:Number of apoptotic cells as well as mRNA contents of Caspase-3 and c-fos of BMSCs group, EPO group and BMSCs+EPO group was lower than those of control group, and number of apoptotic cells as well as mRNA contents of Caspase-3 and c-fos of BMSCs+EPO group were lower than those of BMSCs group and EPO group; mRNA contents of NF-200 and MBP as well as protein contents of NGF and BDNF in spinal cord tissue of BMSCs group, EPO group and BMSCs+EPO group were higher than those of control group, and mRNA contents of NF-200 and MBP as well as protein contents of NGF and BDNF in spinal cord tissue of BMSCs+EPO group were higher than those of BMSCs group and EPO group.Conclusions:Bone marrow mesenchymal stem cells combined with erythropoietin therapy can inhibit cell apoptosis in the injured spinal cord tissue, increase neurotrophic factor levels and inhibit apoptosis and injury molecule expression; it has protective effect on spinal cord injury.

  7. Low power laser and LED irradiation effect on proliferation and differentiation of Wistar rats mesenchymal stem cells

    Science.gov (United States)

    Mancera, Diana; Solarte, Efrain; Fierro, Leonardo; Criollo, William

    2013-11-01

    It has been demonstrated that appropriately cultured and stimulated mesenchymal cells, can give rise to cells of all tissues of the body. We evaluate the cell proliferation and differentiation induced by low power light irradiation in cell cultures of mesenchymal cells, isolated and previously characterized, from Wistar rats. Roche® XTT and LDH tests were used to assess proliferation and cytotoxicity. Cellular differentiation was determined by optical microscopy and using specific fluorescent markers. We report laser cellular proliferation enhancement by 532 and 473 nm, and the best cell culture response by a dose of 2 Jcm-2. Although a three day irradiation protocol the cultures grown and no cytotoxicity was detected. Cellular differentiation occurred, and the production of cardiomyocytes was promoted by the cell proliferation stimulated by low power laser irradiation.

  8. Therapeutic effect of transplanting bone mesenchymal stem cells on the hind limbs’ motor function of rats with acute spinal cord injury

    OpenAIRE

    Song, Qinghua; Xu, Rongmei; Zhang, Quanhai; Ma, Ming; Zhao, Xinping

    2014-01-01

    Purpose: To research the therapeutic effect of the allograft of bone mesenchymal stem cells (BMSCs) on hind limbs’ motor function of rats that underwent acute injury to their spinal nerve. Design: 40 Wistar rat samples with the acute injury to the spinal cord were established and divided into the transplantation group and the control group, 20 for each group; One week after injury, BMSCs were slowly injected into the center of the injured spinal cord of the rats, and the physiological saline ...

  9. Adipose-derived mesenchymal stem cells markedly attenuate brain infarct size and improve neurological function in rats

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    Sun Cheuk-Kwan

    2010-06-01

    Full Text Available Abstract Background The therapeutic effect of adipose-derived mesenchymal stem cells (ADMSCs on brain infarction area (BIA and neurological status in a rat model of acute ischemic stroke (IS was investigated. Methods Adult male Sprague-Dawley (SD rats (n = 30 were divided into IS plus intra-venous 1 mL saline (at 0, 12 and 24 h after IS induction (control group and IS plus intra-venous ADMSCs (2.0 × 106 (treated interval as controls (treatment group after occlusion of distal left internal carotid artery. The rats were sacrificed and brain tissues were harvested on day 21 after the procedure. Results The results showed that BIA was larger in control group than in treatment group (p Conclusions ADMSC therapy significantly limited BIA and improved sensorimotor dysfunction after acute IS.

  10. Repair effect of transplantation of bone marrow mesenchymal stem cells on liver injury in severe burned rats and its mechanism

    International Nuclear Information System (INIS)

    Objective: To investigate the repair effect of transplantation of bone marrow mesenchymal stem cells (BMSCs) on liver injury in severe burned rats, and to clarify its mechanism. Methods: The BMSCs of rats were isolated, cultured, amplified, identified, and labeled in vitro. 30 Wistar rats were randomly divided into normal control group (n=10), model group (n=10) and cell therapy group (n=10). The burned rat model was established. The BMSCs labeled by chlormethyl-benzamidodialkylcarbocyanine (CM-Dil) were transplanted into the rats in cell therapy group by retro-orbital intravenous injection and the saline was injected into the rats in model group. The general status of all rats were observed. The liver tissues of rats were obtained 2 weeks after transplantation, and the pathohistological changes were observed and the pathohistological scores were detected; the apoptotic rate of liver cells was detected by TUNEL method; the engraftment of BMSCs in liver tissues of the rats was observed under laser scanning confocal microscope. Results: 2 weeks after transplantation, the rats in model group were obviously malaise dispirited and the rats in cell therapy group showed obviously better, and the body weight of the rats in cell therapy group was higher than that in model group (P<0.05). The pathohistological results showed the normal liver lobules of the rats in model group disappeared, and the liver cords disordered, and some liver sinusoids dilated and congested, lymphocytes infiltrated with occasional focal aggregating, and cell edema was found, cytoplasm loose and steatosis were seen in liver tissue. However, the pathohistological changes of liver tissue of the rats in cell therapy group were significantly better than those in model group. The pathohistological score of the rats in cell therapy group was significantly lower than that in model group (P<0.05). The TUNEL staining results showed that there were lots of apoptotic liver cells in liver tissue of the rats in

  11. Rat bone marrow mesenchymal stem cells differentiate into hepatocytes in vitro

    Institute of Scientific and Technical Information of China (English)

    Xin-Qin Kang; Wei-Jin Zang; Tu-Sheng Song; Xiao-Li Xu; Xiao-Jiang Yu; Dong-Ling Li; Ke-Wei Meng; Sheng-Li Wu; Zhi-Ying Zhao

    2005-01-01

    AIM: To investigate the mechanism and regulation of differentiation from bone marrow mesenchymal stem cells (MSCs) into hepatocytes and to find a new source of celltypes for therapies of hepatic diseases. METHODS: MSCs were isolated by combining gradient density centrifugation with plastic adherence. The cells were cultured in osteogenic or adipogenic differentiation medium and determined by histochemical staining. MSCs were plated in plastic culture flasks that were not coated with components of extracellular matrix (ECM). When MSCs reached 70% confluence, they were cultured in low glucose Dulbecco's modified Eagle's medium supplemented with 10 mL/L fetal bovine serum, 20 ng/mL hepatocyte growth factor (HGF) and 10 ng/mL fibroblast growth factor-4 (FGF-4). The medium was changed every 3 d and stored for albumin, alpha-fetoprotein (AFP) and urea assay. Glycogen store of hepatocytes was determined by periodic acid-Schiff staining.RESULTS: By combining gradient density centrifugation with plastic adherence, we isolated a homogeneous population of cells from rat bone marrow and differentiated them into osteocytes and adipocytes. When MSCs were cultured withFGF-4 and HGF, approximately 56.6% of cells became smallround and epithelioid on d 24 by morphology. Compared with the control, levels of AFP increased significantly from d 12 to 15.5±1.4 μg/L (t = 2.31, P<0.05) in MSCs cultured with FGF-4and HGF, and were higher (46.2±1.5 μg/L)ond 21 (t = 41.926, P<0.01), then decreased to 24.8±2.2 μg/L on d 24 (t = 10.345, P<0.01). Albumin increased significantly on d 21 (t= 3.325, P<0.01) to 1.4±0.2 μg/mL,and to 2.1±0.7 μg/mL on d 24 (t= 3.646, P<0.01). Urea(2.3±0.4 mmol/L) was first detected on d 21 (t = 6.739, P<0.01), and continued to increase to 2.6±0.9 mmol/Lon d 24 (t= 4.753, P<0.01). Glycogen storage was first seen on d 21.CONCLUSION: The method combining gradient density centrifugation with plastic adherence can isolate MSCs. Rat MSCs may be

  12. Transplantation of Hypoxia Preconditioned Bone Marrow Mesenchymal Stem Cells Enhances Angiogenesis and Neurogenesis after Cerebral Ischemia in Rats

    OpenAIRE

    Wei, Ling; Jamie L. Fraser; Zhong-Yang, Lu; Hu, Xinyang; Yu, Shan Ping

    2012-01-01

    Hypoxic preconditioning of stem cells and neural progenitor cells has been tested for promoting cell survival after transplantation. The present investigation examined the hypothesis that hypoxic preconditioning of bone marrow mesenchymal stem cells (BMSCs) could not only enhance their survival but also reinforce regenerative properties of these cells. BMSCs from eGFP engineered rats or pre-labeled with BrdU were pre-treated with normoxia (20% O2, N-BMSCs) or sublethal hypoxia (0.5% O2. H-BMS...

  13. Use of Rats Mesenchymal Stem Cells Modified with mHCN2 Gene to Create Biologic Pacemakers

    Institute of Scientific and Technical Information of China (English)

    马金; 张存泰; 黄深; 王国强; 全小庆

    2010-01-01

    The possibility of rats mesenchymal stem cells (MSCs) modified with murine hyperpolarization-activated cyclic nucleotide-gated 2 (mHCN2) gene as biological pacemakers in vitro was studied. The cultured MSCs were transfected with pIRES2-EGFP plasmid carrying enhanced green fluorescent protein (EGFP) gene and mHCN2 gene. The identification using restriction enzyme and sequencing indicated that the mHCN2 gene was inserted to the pIRES2-EGFP. Green fluorescence could be seen in MSCs after transfection for 24-48...

  14. Research of Osteo-induced Rat Mesenchymal Stem Cells Seeding on β-TCP/PLLA Porous Composite

    Institute of Scientific and Technical Information of China (English)

    Qiao-Feng WU; Yan-Juan TANG; Huai-Qing CHEN; Jiang WU; Guang-Fu YIN

    2005-01-01

    @@ 1 Introduction Porous biodegradable synthetic materials such as β-tricalcium phosphate (β-TCP) and poly L-lactic acid(PLLA) are usually used in bone tissue engineering.However, proper ratio, optimum pore size and porosity of the composites of β-TCP/PLLA are not very clear yet. In this study, we chose osteo-induced rat bone mesenchymal stem cells (rMSCs) as seeding cells, and cultured them on β-TCP/ PLLA scaffolds with different ratio, different pore size and different porosity. The aim is to investigate the optimum construction for supporting cell growth, proliferation and differentiation.

  15. Cognitive improvement following transvenous adipose-derived mesenchymal stem cell transplantation in a rat model of traumatic brain injury

    Institute of Scientific and Technical Information of China (English)

    Dongfei Li; Chun Yang; Rongmei Qu; Huiying Yang; Meichun Yu; Hui Tao; Jingxing Dai; Lin Yuan

    2011-01-01

    The effects of adipose-derived mesenchymal stem cell (ADMSC) transplantation for the repair of traumatic brain injury remain poorly understood. The present study observed neurological functional changes in a rat model of traumatic brain injury following ADMSC transplantation via the tail vein.Cell transplants were observed in injured cerebral cortex, and expression of brain-derived nerve growth factor was significantly increased in the injured hippocampus following transplantation. Results demonstrated that transvenous ADMSC transplants migrated to the injured cerebral cortex and significantly improved cognitive function.

  16. Collagen-GAG Scaffolds Grafted Onto Myocardial Infarcts in a Rat Model:A Delivery Vehicle for Mesenchymal Stem Cells

    Institute of Scientific and Technical Information of China (English)

    Z.; XIANG; R.; LIAO; M.; KELLY; M.; SPECTOR

    2005-01-01

    1 IntroductionThe objective of the present study was to investigate the response of rat myocardial scar tissue to type I collagen-glycosaminoglycan (GAG) tissue engineering scaffolds, and to assess the feasibility of using a collagen-GAG scaffold as a delivery vehicle for bone marrow-derived mesenchymal stem cells. The benefits of employing the collagen-GAG scaffold for this application include the following:(1) the large surface area of the three-dimensional sponge-like material allows for the delivery of ...

  17. Mesenchymal Stem Cells Derived from Rat Epicardial Versus Epididymal Adipose Tissue

    Directory of Open Access Journals (Sweden)

    Mohamadreza Baghaban Eslaminejad

    2011-01-01

    Full Text Available Objective(sSome investigation has indicated that adipose-derived stem cells possess different surface epitopes and differentiation potential according to the localization of fat pad from which the cells were derived. In the present study proliferation capacity and aging of such cells were explored.Materials and MethodsAdherent cells were isolated from the collagenase digests of adipose tissues excised from rat epicardial and epididymal regions and propagated with several subcultures. The cells were then investigated whether or not they were able to differentiate into bone, cartilage and adipose cell lineages. Studied cells from two adipose tissues were also compared with respect to their in vitro proliferation capacity. The presence of senescent cells in the culture was determined and compared using senescence-associated (SA ß-galactosidase staining method. ResultsSuccessful differentiations of the cells were indicative of their mesenchymal stem cells (MSCs identity. Epicardial adipose-derived cells tended to have a short population doubling time (45±9.6 hr than the epididymal adipose-derived stem cells (69±16 hr, P< 0.05. Colonogenic activity and the growth curve characteristics were all better in the culture of stem cells derived from epicardial compared to epididymal adipose tissue. Comparatively more percentage of senescent cells was present at the cultures derived from epididymal adipose tissue (P< 0.05.ConclusionOur data emphasize on the differences existed between the stem cells derived from adipose depots of different anatomical sites in terms of their proliferative capacity and in vitro aging. Such data can help understand varying results reported by different laboratories involved in adipose stem cell investigations.

  18. Potential Spermatogenesis Recovery with Bone Marrow Mesenchymal Stem Cells in an Azoospermic Rat Model

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    Deying Zhang

    2014-07-01

    Full Text Available Non-obstructive azoospermia is the most challenging type of male infertility. Stem cell based therapy provides the potential to enhance the recovery of spermatogenesis following cancer therapy. Bone marrow-derived mesenchymal stem cells (BMSCs possess the potential to differentiate or trans-differentiate into multi-lineage cells, secrete paracrine factors to recruit the resident stem cells to participate in tissue regeneration, or fuse with the local cells in the affected region. In this study, we tested whether spermatogenically-induced BMSCs can restore spermatogenesis after administration of an anticancer drug. Allogeneic BMSCs were co-cultured in conditioned media derived from cultured testicular Sertoli cells in vitro, and then induced stem cells were transplanted into the seminiferous tubules of a busulfan-induced azoospermatic rat model for 8 weeks. The in vitro induced BMSCs exhibited specific spermatogonic gene and protein markers, and after implantation the donor cells survived and located at the basement membranes of the recipient seminiferous tubules, in accordance with what are considered the unique biological characteristics of spermatogenic stem cells. Molecular markers of spermatogonial stem cells and spermatogonia (Vasa, Stella, SMAD1, Dazl, GCNF, HSP90α, integrinβ1, and c-kit were expressed in the recipient testis tissue. No tumor mass, immune response, or inflammatory reaction developed. In conclusion, BMSCs might provide the potential to trans-differentiate into spermatogenic-like-cells, enhancing endogenous fertility recovery. The present study indicates that BMSCs might offer alternative treatment for the patients with azoospermatic infertility after cancer chemotherapy.

  19. Differentiation potential of bone marrow mesenchymal stem cells into retina in normal and laser-injured rat eye

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jie; SHAN Qing; MA Ping; JIANG Yanming; CHEN Peng; WEN Jingxia; ZHOU You; QIAN Huanwen; PEI Xuetao

    2004-01-01

    Bone marrow mesenchymal stem cells (MSCs) can develop into hematopoietic and mesenchymal lineages but have not been known to participate in the production of retina. Here we report that bone marrow mesenchymal stem cells, after being subretinally transplanted into normal or Nd: YAG laser-injured rat eye, can integrate into RPE layer, photoreceptor layer, bipolar cell layer and ganglion layer. DAPI-labeling detection was used to trace the origin of the repopulating cells. DAPI fluorescence was used to identify retina cells of bone marrow origin 10, 20, 35 and 50 days after transplantation. No formation of rosettes was found but some random cells were found at the end of the observation. MSCs-originated cells spread more widely in the injured retinas than in the normal ones. Immunohistochemical detection showed that though the cells could express neuronal nuclei (NeuN), neuron specific enolase (NSE), glial fibrillary acidic protein (GFAP) and cytokeratin (CK), the proteins expression in the injured transplantation group was abnormal in some region compared with that in the normal transplantation group. Electroretinogram (ERG) showed that ERG-b wave of the injured transplantation group is significantly higher than that of the two laser-injured control groups. These results suggest that a proportion of MSCs can differentiate into retina-like structure in vivo and the differentiation differs in normal and laser-injured retinas.

  20. [Inhibitory effect of total saponins of Panax notoginseng on rat bone marrow mesenchymal stem cell apoptosis].

    Science.gov (United States)

    Ou, Yong-Fang; Fu, Xue-Kun; Mei, Xing-Sha; Zheng, Hui-Zhen

    2016-06-25

    The study was aimed to investigate the effects of total saponins of Panax notoginseng (tPNS) on cobalt chloride (CoCl2)-induced apoptosis of rat bone marrow mesenchymal stem cells (rBMSCs) and the underlying mechanism. rBMSCs were isolated by density gradient centrifugation from Sprague Dawley (SD) rats. After being incubated with different concentrations of tPNS (1, 10, 100 μg/mL) for 48 h, the rBMSCs were stained with EdU and PI for proliferation and cell cycle assay, respectively. CoCl2 group was treated with 300 μmol CoCl2 for 24 h, and different concentrations tPNS groups were treated with 300 μmol CoCl2 plus 1, 10 or 100 μg/mL tPNS. After Annexin V-FITC/PI staining, flow cytometry was applied to measure the cell apoptosis. For mitochondrial membrane potential assay, rhodamine123 and Hoechst33342 staining were used. qRT-PCR was applied to analyze gene expression of Bcl-2 family. The results showed that the proliferation rates of the three concentrations tPNS groups were all higher than that of the control group (all P < 0.05). Compared with control group, only 100 μg/mL tPNS group exhibited increased cell percentage of S and G2 phase. Compared with that in control group (without CoCl2), the apoptotic rate was increased by 14.2% in CoCl2 group. And the apoptotic rates were reduced by 14.4%, 12.8% and 13.9% in three concentrations tPNS groups, compared with that in CoCl2 group (all P < 0.01). CoCl2 could decrease the mitochondrial membrane potential, while different concentrations of tPNS reversed the inhibitory effect of CoCl2. Bcl-2 and Bcl-xl mRNA expressions in all tPNS groups were higher than those in CoCl2 group (all P < 0.05). Moreover, 10 and 100 μg/mL tPNS groups showed lower ratios of Bax/Bcl-2, compared with CoCl2 group. The results suggest that tPNS protects the rBMSCs against CoCl2-induced apoptosis through improving the cell mitochondrial membrane potential, up-regulating the expressions of anti-apoptosis genes Bcl-2 and Bcl-xl, and reducing

  1. Effects of strontium on proliferation and differentiation of rat bone marrow mesenchymal stem cells

    Energy Technology Data Exchange (ETDEWEB)

    Li, Yunfeng; Li, Jihua; Zhu, Songsong; Luo, En; Feng, Ge; Chen, Qianming [State Key Laboratory of Oral Diseases, West China College of Stomatology, Sichuan University, No. 14, Section 3, Southern Renmin Road, Chengdu 610041 (China); Hu, Jing, E-mail: drhu@vip.sohu.com [State Key Laboratory of Oral Diseases, West China College of Stomatology, Sichuan University, No. 14, Section 3, Southern Renmin Road, Chengdu 610041 (China)

    2012-02-24

    Highlights: Black-Right-Pointing-Pointer Strontium ranelate (SrR) inhibits proliferation of BMMSCs. Black-Right-Pointing-Pointer SrR increases osteoblastic but decreases adipocytic differentiation of BMMSCs. Black-Right-Pointing-Pointer SrR increases expression of Runx2, BSP and OCN by BMMSCs in osteogenic medium. Black-Right-Pointing-Pointer SrR decreases expression of PPAR{gamma}, aP2/ALBP and LPL by BMMSCs in adipogenic medium. -- Abstract: Strontium ranelate (SrR) was an effective anti-osteoporotic drug to increase bone formation and decrease bone resorption. However, reports about the effect of SR on osteoblastic and adipocytic differentiation from bone marrow mesenchymal stem cells (BMMSCs) are limited. The purpose of this study is to evaluate whether SrR affects the ability of BMMSCs to differentiate into osteoblasts or adipocytes. Rat BMMSCs were identified by flow cytometry and exposed to SR (0.1 and 1.0 mM Sr{sup 2+}) under osteogenic or adipogenic medium for 1 and 2 weeks. The proliferation and differentiation of BMMSCs were analyzed by MTT, alkaline phosphatase (ALP), Oil red O staining, quantitative real-time RT-PCR and Western blot assays. SrR significantly inhibited the proliferation, increased osteoblastic but decreased adipocytic differentiation of rat BMMSCs dose-dependently. In osteogenic medium, SrR increased the expression of ALP, the mRNA levels of Cbfa1/Runx2, bone sialoprotein, and osteocalcin by RT-PCR, and the protein levels of Cbfa1/Runx2 by Western blot. In adipogenic medium, SrR decreased the mRNA levels of PPAR{gamma}2, adipocyte lipid-binding protein 2 (aP2/ALBP), and lipoprotein lipase (LPL) by RT-PCR, and the protein expression of PPAR{gamma} in Western blot analysis. These results indicated that the effects of SrR to promote osteoblastic but inhibit adipocytic differentiation of BMMSCs might contribute to its effect on osteoporosis treatment.

  2. Human umbilical cord mesenchymal stem cells reduce systemic inflammation and attenuate LPS-induced acute lung injury in rats

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    Li Jianjun

    2012-09-01

    Full Text Available Abstract Background Mesenchymal stem cells (MSCs possess potent immunomodulatory properties and simultaneously lack the ability to illicit immune responses. Hence, MSCs have emerged as a promising candidate for cellular therapeutics for inflammatory diseases. Within the context of this study, we investigated whether human umbilical cord-derived mesenchymal stem cells (UC-MSCs could ameliorate lipopolysaccharide- (LPS- induced acute lung injury (ALI in a rat model. Methods ALI was induced via injection of LPS. Rats were divided into three groups: (1 saline group(control, (2 LPS group, and (3 MSC + LPS group. The rats were sacrificed at 6, 24, and 48 hours after injection. Serum, bronchoalveolar lavage fluid (BALF, and lungs were collected for cytokine concentration measurements, assessment of lung injury, and histology. Results UC-MSCs increased survival rate and suppressed LPS-induced increase of serum concentrations of pro-inflammatory mediators TNF-α, IL-1β, and IL-6 without decreasing the level of anti-inflammatory cytokine IL-10. The MSC + LPS group exhibited significant improvements in lung inflammation, injury, edema, lung wet/dry ratio, protein concentration, and neutrophil counts in the BALF, as well as improved myeloperoxidase (MPO activity in the lung tissue. Furthermore, UC-MSCs decreased malondialdehyde (MDA production and increased Heme Oxygenase-1 (HO-1 protein production and activity in the lung tissue. Conclusion UC-MSCs noticeably increased the survival rate of rats suffering from LPS-induced lung injury and significantly reduced systemic and pulmonary inflammation. Promoting anti-inflammatory homeostasis and reducing oxidative stress might be the therapeutic basis of UC-MSCs.

  3. In Vivo Tracking of Systemically Administered Allogeneic Bone Marrow Mesenchymal Stem Cells in Normal Rats through Bioluminescence Imaging

    Science.gov (United States)

    Cao, Juan; Hou, Shike; Ding, Hui; Liu, Ziquan; Song, Meijuan; Qin, Xiaojing; Wang, Xue; Yu, Mengyang; Sun, Zhiguang; Liu, Jinyang; Sun, Shuli; Xiao, Peixin

    2016-01-01

    Recently, mesenchymal stem cells (MSCs) are increasingly used as a panacea for multiple types of disease short of effective treatment. Dozens of clinical trials published demonstrated strikingly positive therapeutic effects of MSCs. However, as a specific agent, little research has focused on the dynamic distribution of MSCs after in vivo administration. In this study, we track systemically transplanted allogeneic bone marrow mesenchymal stem cells (BMSCs) in normal rats through bioluminescence imaging (BLI) in real time. Ex vivo organ imaging, immunohistochemistry (IHC), and RT-PCR were conducted to verify the histological distribution of BMSCs. Our results showed that BMSCs home to the dorsal skin apart from the lungs and kidneys after tail vein injection and could not be detected 14 days later. Allogeneic BMSCs mainly appeared not at the parenchymatous organs but at the subepidermal connective tissue and adipose tissue in healthy rats. There were no significant MSCs-related adverse effects except for transient decrease in neutrophils. These findings will provide experimental evidences for a better understanding of the biocharacteristics of BMSCs.

  4. In Vivo Tracking of Systemically Administered Allogeneic Bone Marrow Mesenchymal Stem Cells in Normal Rats through Bioluminescence Imaging.

    Science.gov (United States)

    Cao, Juan; Hou, Shike; Ding, Hui; Liu, Ziquan; Song, Meijuan; Qin, Xiaojing; Wang, Xue; Yu, Mengyang; Sun, Zhiguang; Liu, Jinyang; Sun, Shuli; Xiao, Peixin; Lv, Qi; Fan, Haojun

    2016-01-01

    Recently, mesenchymal stem cells (MSCs) are increasingly used as a panacea for multiple types of disease short of effective treatment. Dozens of clinical trials published demonstrated strikingly positive therapeutic effects of MSCs. However, as a specific agent, little research has focused on the dynamic distribution of MSCs after in vivo administration. In this study, we track systemically transplanted allogeneic bone marrow mesenchymal stem cells (BMSCs) in normal rats through bioluminescence imaging (BLI) in real time. Ex vivo organ imaging, immunohistochemistry (IHC), and RT-PCR were conducted to verify the histological distribution of BMSCs. Our results showed that BMSCs home to the dorsal skin apart from the lungs and kidneys after tail vein injection and could not be detected 14 days later. Allogeneic BMSCs mainly appeared not at the parenchymatous organs but at the subepidermal connective tissue and adipose tissue in healthy rats. There were no significant MSCs-related adverse effects except for transient decrease in neutrophils. These findings will provide experimental evidences for a better understanding of the biocharacteristics of BMSCs. PMID:27610137

  5. Spirolactone provides protection from renal fibrosis by inhibiting the endothelial–mesenchymal transition in isoprenaline-induced heart failure in rats

    OpenAIRE

    Zhou, Hao; Xi, Dan; Liu, Jichen; Zhao, Jinjin; Chen, Si; Guo, Zhigang

    2016-01-01

    Background Fibrosis results in excessive accumulation of extracellular matrix proteins, collagen component alteration, and abnormalities in structure and is partly derived from a process called the endothelial–mesenchymal transition involving transforming growth factor β (TGF-β). We investigated whether spironolactone, an aldosterone receptor blocker, attenuated isoprenaline (Iso)-induced heart failure in rats and also studied the mechanism for the same. Methods Sprague–Dawley rats were subcu...

  6. Mesenchymal stem cells from rats with chronic kidney disease exhibit premature senescence and loss of regenerative potential.

    Directory of Open Access Journals (Sweden)

    Barbara Mara Klinkhammer

    Full Text Available Mesenchymal stem cell (MSC transplantation has the potential for organ repair. Nevertheless, some factors might lessen the regenerative potential of MSCs, e.g. donor age or systemic disease. It is thus important to carefully assess the patient's suitability for autologous MSC transplantation. Here we investigated the effects of chronic kidney disease (CKD on MSC function. We isolated bone marrow MSCs from remnant kidney rats (RK with CKD (CKD-RK-MSC and found signs of premature senescence: spontaneous adipogenesis, reduced proliferation capacity, active senescence-associated-β-galactosidase, accumulation of actin and a modulated secretion profile. The functionality of CKD-RK-MSCs in vivo was tested in rats with acute anti-Thy1.1-nephritis, where healthy MSCs have been shown to be beneficial. Rats received healthy MSCs, CKD-RK-MSC or medium by injection into the left renal artery. Kidneys receiving healthy MSCs exhibited accelerated healing of glomerular lesions, whereas CKD-RK-MSC or medium exerted no benefit. The negative influence of advanced CKD/uremia on MSCs was confirmed in a second model of CKD, adenine nephropathy (AD. MSCs from rats with adenine nephropathy (CKD-AD-MSC also exhibited cellular modifications and functional deficits in vivo. We conclude that CKD leads to a sustained loss of in vitro and in vivo functionality in MSCs, possibly due to premature cellular senescence. Considering autologous MSC therapy in human renal disease, studies identifying uremia-associated mechanisms that account for altered MSC function are urgently needed.

  7. Human embryonic mesenchymal stem cell-derived conditioned medium rescues kidney function in rats with established chronic kidney disease.

    Directory of Open Access Journals (Sweden)

    Arianne van Koppen

    Full Text Available Chronic kidney disease (CKD is a major health care problem, affecting more than 35% of the elderly population worldwide. New interventions to slow or prevent disease progression are urgently needed. Beneficial effects of mesenchymal stem cells (MSC have been described, however it is unclear whether the MSCs themselves or their secretome is required. We hypothesized that MSC-derived conditioned medium (CM reduces progression of CKD and studied functional and structural effects in a rat model of established CKD. CKD was induced by 5/6 nephrectomy (SNX combined with L-NNA and 6% NaCl diet in Lewis rats. Six weeks after SNX, CKD rats received either 50 µg CM or 50 µg non-CM (NCM twice daily intravenously for four consecutive days. Six weeks after treatment CM administration was functionally effective: glomerular filtration rate (inulin clearance and effective renal plasma flow (PAH clearance were significantly higher in CM vs. NCM-treatment. Systolic blood pressure was lower in CM compared to NCM. Proteinuria tended to be lower after CM. Tubular and glomerular damage were reduced and more glomerular endothelial cells were found after CM. DNA damage repair was increased after CM. MSC-CM derived exosomes, tested in the same experimental setting, showed no protective effect on the kidney. In a rat model of established CKD, we demonstrated that administration of MSC-CM has a long-lasting therapeutic rescue function shown by decreased progression of CKD and reduced hypertension and glomerular injury.

  8. Salt-induced epithelial-to-mesenchymal transition in Dahl salt-sensitive rats is dependent on elevated blood pressure

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Y.; Mu, J.J.; Liu, F.Q.; Ren, K.Y.; Xiao, H.Y. [Xi' an Jiaotong University, Medical College, First Affiliated Hospital, Cardiovascular Department, Xi' an, China, Cardiovascular Department, First Affiliated Hospital, Medical College, Xi' an Jiaotong University, Xi' an (China); Ministry of Education, Key Laboratory of Environment and Genes Related to Diseases, Xi' an, China, Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Xi' an (China); Yang, Z. [Xi' an Jiaotong University, Medical College, First Affiliated Hospital, Department of Pathology, Xi' an, China, Department of Pathology, First Affiliated Hospital, Medical College, Xi' an Jiaotong University, Xi' an (China); Yuan, Z.Y. [Xi' an Jiaotong University, Medical College, First Affiliated Hospital, Cardiovascular Department, Xi' an, China, Cardiovascular Department, First Affiliated Hospital, Medical College, Xi' an Jiaotong University, Xi' an (China); Ministry of Education, Key Laboratory of Environment and Genes Related to Diseases, Xi' an, China, Key Laboratory of Environment and Genes Related to Diseases, Ministry of Education, Xi' an (China)

    2014-03-03

    Dietary salt intake has been linked to hypertension and cardiovascular disease. Accumulating evidence has indicated that salt-sensitive individuals on high salt intake are more likely to develop renal fibrosis. Epithelial-to-mesenchymal transition (EMT) participates in the development and progression of renal fibrosis in humans and animals. The objective of this study was to investigate the impact of a high-salt diet on EMT in Dahl salt-sensitive (SS) rats. Twenty-four male SS and consomic SS-13{sup BN} rats were randomized to a normal diet or a high-salt diet. After 4 weeks, systolic blood pressure (SBP) and albuminuria were analyzed, and renal fibrosis was histopathologically evaluated. Tubular EMT was evaluated using immunohistochemistry and real-time PCR with E-cadherin and alpha smooth muscle actin (α-SMA). After 4 weeks, SBP and albuminuria were significantly increased in the SS high-salt group compared with the normal diet group. Dietary salt intake induced renal fibrosis and tubular EMT as identified by reduced expression of E-cadherin and enhanced expression of α-SMA in SS rats. Both blood pressure and renal interstitial fibrosis were negatively correlated with E-cadherin but positively correlated with α-SMA. Salt intake induced tubular EMT and renal injury in SS rats, and this relationship might depend on the increase in blood pressure.

  9. Salt-induced epithelial-to-mesenchymal transition in Dahl salt-sensitive rats is dependent on elevated blood pressure

    International Nuclear Information System (INIS)

    Dietary salt intake has been linked to hypertension and cardiovascular disease. Accumulating evidence has indicated that salt-sensitive individuals on high salt intake are more likely to develop renal fibrosis. Epithelial-to-mesenchymal transition (EMT) participates in the development and progression of renal fibrosis in humans and animals. The objective of this study was to investigate the impact of a high-salt diet on EMT in Dahl salt-sensitive (SS) rats. Twenty-four male SS and consomic SS-13BN rats were randomized to a normal diet or a high-salt diet. After 4 weeks, systolic blood pressure (SBP) and albuminuria were analyzed, and renal fibrosis was histopathologically evaluated. Tubular EMT was evaluated using immunohistochemistry and real-time PCR with E-cadherin and alpha smooth muscle actin (α-SMA). After 4 weeks, SBP and albuminuria were significantly increased in the SS high-salt group compared with the normal diet group. Dietary salt intake induced renal fibrosis and tubular EMT as identified by reduced expression of E-cadherin and enhanced expression of α-SMA in SS rats. Both blood pressure and renal interstitial fibrosis were negatively correlated with E-cadherin but positively correlated with α-SMA. Salt intake induced tubular EMT and renal injury in SS rats, and this relationship might depend on the increase in blood pressure

  10. Autologous adipose tissue‑derived mesenchymal stem cells are involved in rat liver regeneration following repeat partial hepatectomy.

    Science.gov (United States)

    Liu, Tao; Mu, Hong; Shen, Zhongyang; Song, Zhuolun; Chen, Xiaobo; Wang, Yuliang

    2016-03-01

    Adipose tissue‑derived mesenchymal stem cells (ADSCs) have been considered to be attractive and readily available adult mesenchymal stem cells, and they are becoming increasingly popular for use in regenerative cell therapy, as they are readily accessible through minimally invasive techniques. The present study investigated whether autologous ADSC transplantation promoted liver regeneration following a repeat partial hepatectomy in rats. The rats were divided into three groups as follows: 70% partial hepatectomy (PH) group; repeat PH (R‑PH) group and R‑PH/ADSC group, subjected to R‑PH and treated with autologous ADSCs via portal vein injection. In each group, the rats were sacrificed at different time points postoperatively in order to evaluate the changes in liver function and to estimate the liver regenerative response. The expression of proliferating cell nuclear antigen (PCNA) labeling index in the liver was measured using immunohistochemistry. The expression levels of hepatocyte growth factor (HGF) mRNA were measured using reverse transcription polymerase chain reaction. The results showed that regeneration of the remaining liver following R‑PH was significantly promoted by ADSC transplantation, as shown by a significant increase in liver to body weight ratio and the PCNA labeling index at 24 h post‑hepatectomy. Additionally, ADSC transplantation markedly inhibited the elevation of serum levels of alanine aminotransferase, aspartate aminotransferase and total bilirubin, increased HGF content and also attenuated hepatic vacuolar degeneration 24 h postoperatively. Furthermore, the liver was found to almost fully recover from hepatocellular damage due to hepatectomy among the three groups at 168 h postoperatively. These results indicated that autologous ADSC transplantation enhanced the regenerative capacity of the remnant liver tissues in the early phase following R‑PH. PMID:26783183

  11. Effect of mesenchymal stem cells transplantation on glycaemic profile & their localization in streptozotocin induced diabetic Wistar rats

    Directory of Open Access Journals (Sweden)

    Shobhit Bhansali

    2015-01-01

    Full Text Available Background & objectives: Bone marrow is a rich source of adult stem cells that can differentiate into various cell types. Administration of mesenchymal stem cells (MSCs in irradiated diabetic rat model has transiently shown to decrease blood glucose level. This study examines the effect of high dose and multiple injections of MSCs on glycemic profile, their localization and regeneration of islet in diabetic Wistar rat. Methods: The study was carried out in male Wistar rats categorized into three groups (n=6, in each group: Group 1 as control, group 2 streptozotocin (STZ (50 mg/kg induced diabetic group and group 3 experimental group; 5-bromo-2-deoxyuridine (BrdU labelled allogenic MSCs were injected in the non-irradiated diabetic rat of the experimental group through tail vein. The blood glucose profile was subsequently monitored at regular intervals. Rats were sacrificed on day 45 and pancreas was examined for localization of BrdU labelled stem cells by immunofluorescence and islet-neogenesis by immunohistochemistry . Results: There was a significant reduction in blood glucose level after administration of MSCs in the experimental group (P<0.001. The presence of BrdU labelled MSCs in islet suggested their localization in the pancreas. Co-expression of anti-BrdU and anti-insulin antibody indicated trans-differentiation / fusion into insulin producing cells evidenced by significant increase in total number of islet (P=0.004 and insulin positive cells ( P<0.0001 in experimental group. Interpretation & conclusions: Our results showed that the MSCs administration in non-irradiated diabetic Wistar rat reduced hyperglycaemia and was accompanied by increased islet-neogengesis, possibly through trans- differentiation/fusion.

  12. Increased stromal-cell-derived factor 1 enhances the homing of bone marrow derived mesenchymal stem cells in dilated cardiomyopathy in rats

    Institute of Scientific and Technical Information of China (English)

    ZHOU Yan-li; Michael Fu; ZHANG Hai-feng; LI Xin-li; DI Ruo-min; YAO Wen-ming; LI Dian-fu; FENG Jian-lin; HUANG Jun; CAO Ke-jiang

    2010-01-01

    Background Stem cell transplantation has been shown to have beneficial effects on dilated cardiomyopathy. However,mechanism for stem cell homing to cardiac tissue in dilated cardiomyopathy has not yet been elucidated.Methods Mesenchymal stem cells were obtained from rat bone marrow, expanded in vitro, and labeled with 99mTc.Cardiomyopathy model was induced by doxorubicin in rats. 99mTc labeled cells were infused into the left ventricles in cardiomyopathy and control rats. Sixteen hours after injection, animals were sacrificed and different tissues were harvested to measure specific radioactivity. By use of real-time polymerase chain reaction and immunohistochemistry,Mrna and protein expressions for stromal-cell-derived factor 1 in cardiac tissue were measured.Results Labeling efficiency of mesenchymal stem cells was (70.0±11.2)%. Sixteen hours after mesenchymal stem cell transplantation, the heart-to-muscle radioactivity ratio was increased significantly in cardiomyopathy hearts as compared to control hearts. Both Mrna and rotein expressions of stromal-cell-derived factor 1 were up-regulated in cardiomyopathy hearts as compared with control hearts.Conclusion In dilated cardiomyopathy induced by doxorubicin up-regulated expression of stromal-cell-derived factor 1in heart may induce mesenchymal stem cells home to the heart.

  13. Freeze-Dried Rat Bone Marrow Mesenchymal Stem Cell Paracrine Factors: A Simplified Novel Material for Skin Wound Therapy

    Science.gov (United States)

    Peng, Yan; Xuan, Min; Zou, Jiping; Liu, Hongwei; Zhuo, Ziyuan; Wan, Yu

    2015-01-01

    The mesenchymal stem cell (MSC) supernatant is well known as a rich source of autologous cytokines and universally used for tissue regeneration in current clinical medicine. However, the limitation of conditioned medium used in open-wound repair compels the need to find a more sophisticated way to take advantage of the trophic factors of MSCs. We have now fabricated a three-dimensional membrane from freeze-dried bone marrow mesenchymal stem cells-conditioned medium (FBMSC-CM) using a simple freeze-dried protocol. Scanning electron microscopy images showed the microstructure of the FBMSC-CM membrane (FBMSC-CMM) resembling a mesh containing growth factors. ELISA was used to test the paracrine factors retained in the FBMSC-CMM, and the results indicated that FBMSC-CMM withheld over 80% of the paracrine factors. Live/dead assays were adopted to test the toxicity of the FBMSC-CMM on cultured rat dermal fibroblasts, and the results confirmed its biological safety with low toxicity. Moreover, the FBMSC-CMM could significantly accelerate wound healing and enhance the neovascularization as well as epithelialization through strengthening the trophic factors in the wound bed as determined by immunohistochemical staining. Thus, the ability to maintain paracrine factors and enhance the effectiveness of these growth factors in the wound as well as the simple procedure and economical materials required for production qualifies the FBMSC-CMM to be a candidate biomaterial for open-wound regeneration. PMID:25343727

  14. Human multipotent mesenchymal stem cells improve healing after collagenase tendon injury in the rat

    Czech Academy of Sciences Publication Activity Database

    Machová-Urdzíková, Lucia; Sedláček, R.; Suchý, T.; Amemori, Takashi; Růžička, Jiří; Lesný, P.; Havlas, V.; Syková, Eva; Jendelová, Pavla

    2014-01-01

    Roč. 13, č. 42 (2014). ISSN 1475-925X R&D Projects: GA ČR GAP304/10/0326; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:68378041 Keywords : Achilles tendon * mesenchymal stromal cells * osteogenesis Subject RIV: FI - Traumatology, Orthopedics Impact factor: 1.427, year: 2014

  15. Airway Delivery of Mesenchymal Stem Cells Prevents Arrested Alveolar Growth in Neonatal Lung Injury in Rats

    OpenAIRE

    van Haaften, Timothy; Byrne, Roisin; Bonnet, Sebastien; Rochefort, Gael Y.; Akabutu, John; Bouchentouf, Manaf; Rey-Parra, Gloria J.; Galipeau, Jacques; Haromy, Alois; Eaton, Farah; Chen, Ming; Hashimoto, Kyoko; Abley, Doris; Korbutt, Greg; Archer, Stephen L.

    2009-01-01

    Rationale: Bronchopulmonary dysplasia (BPD) and emphysema are characterized by arrested alveolar development or loss of alveoli; both are significant global health problems and currently lack effective therapy. Bone marrow–derived mesenchymal stem cells (BMSCs) prevent adult lung injury, but their therapeutic potential in neonatal lung disease is unknown.

  16. The effects of bone marrow mesenchymal stromal cells transplantation after mannitol pretreatment on behavioral performance and synaptophysin expression in the CA3 region in hippocampus of vascular dementia rats

    Institute of Scientific and Technical Information of China (English)

    农伟东

    2013-01-01

    Objective To investigate the effects of bone marrow mesenchymal stromal cells (BMSCs) transplantation after mannitol pretreatment on behavioral performance and synaptophysin expression in the CA3region in hippocampus of vascular dementia (VD) rats.Methods The

  17. The effect of magnetic field during freezing and thawing of rat bone marrow-derived mesenchymal stem cells.

    Science.gov (United States)

    Shikata, H; Kaku, M; Kojima, S-I; Sumi, H; Kojima, S-T; Yamamoto, T; Yashima, Y; Kawata, T; Tanne, K; Tanimoto, K

    2016-08-01

    Previous studies showed that a programmed freezer with magnetic field can maintain a high survival rate of mesenchymal stem cells (MSCs). The purpose of this study was to evaluate the influences of magnetic field during freezing and thawing on the survival of MSCs isolated from rat bone marrow. The cells were frozen by a normal programmed freezer or a programmed freezer with magnetic field (CAS-LAB1) and cryopreserved for 7 days at -150 °C. Then, the cells were thawed in the presence or absence of magnetic field. Immediately after thawing, the number of surviving or viable cells was counted. The cell proliferation was examined after 1-week culture. Cryopreserved MSCs which were frozen by a normal freezer or a CAS freezer were transplanted into bone defects artificially made in calvaria of 4-week-old rats. Non-cryopreserved MSCs were used as a control. The rats were sacrificed at 8, 16, or 24 weeks after transplantation and the bone regeneration area was measured. Proliferation rates of MSCs after 1 week were significantly higher in the CAS-freezing-thawing group than in the CAS-freezing group. The extent of new bone formation in the CAS-freezing-thawing group tended to be larger than in CAS-freezing group 24 weeks after transplantation. These results suggest that a magnetic field enhances cell survival during thawing as well as freezing. PMID:27346603

  18. Synergistic effects of low-level laser and mesenchymal stem cells on functional recovery in rats with crushed sciatic nerves.

    Science.gov (United States)

    Yang, Chen-Chia; Wang, John; Chen, Shyh-Chang; Hsieh, Yueh-Ling

    2016-02-01

    Transplantation of mesenchymal stem cells (MSCs) has been proposed to exert beneficial effects on peripheral nerve regeneration after a peripheral nerve injury, but the functional recovery in the denervated limb is still limited. In this study, we used low-level laser therapy (LLLT) as an adjunct therapy for MSC transplantation on the functional recovery of crushed sciatic nerve in rats. Peripheral nerve injury was induced in 48 Sprague-Dawley rats by crushing the unilateral sciatic nerve, using a vessel clamp. The animals with crushed injury were randomly divided into four groups: control group, with no treatment; MSC group, treated with MSC alone; LLLT group, treated with LLLT alone; and MSCLLLT group, treated with a combination of MSC and LLLT. The sciatic function index (SFI), vertical activity of locomotion (VA) and ankle angle (AA) of rats were examined for functional assessments after treatment. Electrophysiological, morphological and S100 immunohistochemical studies were also conducted. The MSCLLLT group showed a greater recovery in SFI, VA and AA, with significant difference from MSC, LLLT and control groups (p crush injury in the MSCLLLT group when compared with the groups receiving a single treatment (p crush injury. PMID:23468370

  19. In vitro cultivation of rat bone marrow mesenchymal stem cells and establishment of pEGFP/Ang-1 transfection method

    Institute of Scientific and Technical Information of China (English)

    Xiu-Qun; Zhang; Long; Wang; Shu-Li; Zhao; Wei; Xu

    2014-01-01

    Objective:To obtain the bone marrow mesenchymal stem cells(BMSCs).complete phenotypic identification and successfully transfecl rat BMSCs by recombinant plasmid pF.GFP/Ang-1.Methods:BMSCs were isolated from bone marrow using density gradient centrifugation method and adherence screening method,and purified.Then the recombinant plasmid pEGFP/Ang-1was used to transfect BMSCs and the positive clones were obtained by the screen of C418 and observed under light microscopy inversely.Green fluorescent exhibited by protein was enhanced to measure the change time of the expression amount of Ang-1.Results:BMSCs cell lines were obtained successfully by adherence screening method and density gradient ccntrifugation.Ang-1 recombinant plasmid was transfected smoothly into rat BMSCs,which can express Ang-1 for 3 d and decreased after 7 d.Conclusions:Adherence screening method und density gradient ceiilrifugation can be effective methods lo obtain BMSCs with high purity and rapid proliferation.Besides,the expression of transfected recombinant plasmid pEGFP/Ang-1 in rat BMSCs is satisfactory.

  20. Bone marrow mesenchymal stem cells protect against retinal ganglion cell loss in aged rats with glaucoma

    OpenAIRE

    Hu Y; Tan HB; Wang XM; Rong H; Cui HP; Cui H

    2013-01-01

    Ying Hu,1,2 Hai Bo Tan,1 Xin Mei Wang,3 Hua Rong,1 Hong Ping Cui,1 Hao Cui2 Departments of Ophthalmology, 1Shanghai East Hospital of Tongji University, Shanghai, 2First Affiliated Hospital, 3Fourth Affiliated Hospital, Harbin Medical University, Harbin, People's Republic of China Abstract: Glaucoma is a common eye disease in the aged population and has severe consequences. The present study examined the therapeutic effects of bone marrow mesenchymal stem cell (BMSC) transplantation i...

  1. Pre-differentiation of mesenchymal stromal cells in combination with a microstructured nerve guide supports peripheral nerve regeneration in the rat sciatic nerve model.

    Science.gov (United States)

    Boecker, Arne Hendrik; van Neerven, Sabien Geraldine Antonia; Scheffel, Juliane; Tank, Julian; Altinova, Haktan; Seidensticker, Katrin; Deumens, Ronald; Tolba, Rene; Weis, Joachim; Brook, Gary Anthony; Pallua, Norbert; Bozkurt, Ahmet

    2016-02-01

    Many bioartificial nerve guides have been investigated pre-clinically for their nerve regeneration-supporting function, often in comparison to autologous nerve transplantation, which is still regarded as the current clinical gold standard. Enrichment of these scaffolds with cells intended to support axonal regeneration has been explored as a strategy to boost axonal regeneration across these nerve guides Ansselin et al. (1998). In the present study, 20 mm rat sciatic nerve defects were implanted with a cell-seeded microstructured collagen nerve guide (Perimaix) or an autologous nerve graft. Under the influence of seeded, pre-differentiated mesenchymal stromal cells, axons regenerated well into the Perimaix nerve guide. Myelination-related parameters, like myelin sheath thickness, benefitted from an additional seeding with pre-differentiated mesenchymal stromal cells. Furthermore, both the number of retrogradely labelled sensory neurons and the axon density within the implant were elevated in the cell-seeded scaffold group with pre-differentiated mesenchymal stromal cells. However, a pre-differentiation had no influence on functional recovery. An additional cell seeding of the Perimaix nerve guide with mesenchymal stromal cells led to an extent of functional recovery, independent of the differentiation status, similar to autologous nerve transplantation. These findings encourage further investigations on pre-differentiated mesenchymal stromal cells as a cellular support for peripheral nerve regeneration. PMID:26296589

  2. Effects of Nitric Oxide Production Inhibitor Named, NG-Nitro-L-Arginine Methyl Ester (L-NAME, on Rat Mesenchymal Stem Cells Differentiation

    Directory of Open Access Journals (Sweden)

    E Arfaei

    2010-04-01

    Full Text Available Introduction & Objectives: Recently, the findings of some studies have shown that, nitric oxide (NO probably has an important role in differentiation of mesenchymal stem cells to osteoblasts. The aim of the present investigation was to study the effects of nitric oxide production inhibitor named, NG-nitro-L-arginine methyl ester (L-NAME, on rat mesenchymal stem cells differentiation to osteoblasts in vitro. Materials & Methods: This was an experimental study conducted at Hamedan University of Medical Sciences in 2009, in which rat bone marrow stem cells were isolated in an aseptic condition and cultured in vitro. After third passage, the cells were cultured in osteogenic differentiation medium. To study the effects of L-NAME on osteogenic differentiation, the L-NAME was added to the culture medium at a concentration of 125, 250, and 500 μM in some culture plates. During the culture procedure, the media were replaced with fresh ones, with a three days interval. After 28 days of culturing the mineralized matrix was stained using Alizarian red staining method. The gathered data were analyzed by SPSS software version 12 using one way ANOVA. Results: The findings of this study showed that in the presence of L-NAME, differentiation of bone marrow mesenchymal stem cells to osteoblasts was disordered and matrix mineralization significantly decreased in a dose dependent manner. Conclusion: This study revealed that, inhibition of nitric oxide production using L-NAME can prevent the differentiation of rat bone marrow mesenchymal stem cells to osteoblast. The results imply that NO is an important constituent in differentiation of mesenchymal stem cell to osteoblasts.

  3. Effects of mesenchymal stem cells on interleukin-1β-treated chondrocytes and cartilage in a rat osteoarthritic model.

    Science.gov (United States)

    Tang, Jilei; Cui, Weiding; Song, Fanglong; Zhai, Chenjun; Hu, Hansheng; Zuo, Qiang; Fan, Weimin

    2015-08-01

    In the present study, the effects and mechanisms of mesenchymal stem cells (MSCs) on interleukin (IL)-1β-stimulated rat chondrocytes, as well as cartilage from a rat model of osteoarthritis (OA) induced by anterior cruciate ligament transection and medial meniscectomy were investigated. Confluent rat chondrocytes were treated with IL-1β (10 ng/ml), then cultured indirectly with or without MSCs at a ratio of 2:1. Total RNA and protein were collected at various time-points, and western blot and reverse transcription-quantitative polymerase chain reaction analyses were used to investigate the expression of type II collagen (Col2), aggrecan, matrix metalloproteinase-13 (MMP-13) and cyclooxygenase-2 (COX-2). The activation of extracellular signal-regulated kinases 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase (MAPK), nuclear factor-κB (NF-κB) p65 and inhibitory-κ-B-α (IκBα) were also assessed by western blotting. In addition, the in vivo effects of MSCs in a rat OA model were assessed by histology and western blot analysis. The results indicated that in vitro, IL-1β markedly upregulated the expression of MMP-13, COX-2, phosphorylated ERK1/2, JNK, p38 MAPK and NF-κB p65, and inhibited the expression of Col2, aggrecan and IκBα. Conversely, MSCs enhanced the expression of Col2, aggrecan and IκBα, and inhibited the expression of MMP-13 and NF-κB p65 in IL-1β-stimulated rat chondrocytes. In vivo histological and western blot analyses revealed analogous results to the in vitro findings. The results of the present study demonstrated that MSCs suppressed the inflammatory response and extracellular matrix degradation in IL-1β‑induced rat chondrocytes, as well as cartilage in a osteoarthritic rat model, in part via the NF-κB signaling pathway. PMID:25892273

  4. Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide

    Directory of Open Access Journals (Sweden)

    Naaldijk Yahaira

    2012-08-01

    Full Text Available Abstract Background Mesenchymal stem cells (MSCs are increasingly used as therapeutic agents as well as research tools in regenerative medicine. Development of technologies which allow storing and banking of MSC with minimal loss of cell viability, differentiation capacity, and function is required for clinical and research applications. Cryopreservation is the most effective way to preserve cells long term, but it involves potentially cytotoxic compounds and processing steps. Here, we investigate the effect of decreasing dimethyl sulfoxide (DMSO concentrations in cryosolution by substituting with hydroxyethyl starch (HES of different molecular weights using different freezing rates. Post-thaw viability, phenotype and osteogenic differentiation capacity of MSCs were analysed. Results The study confirms that, for rat MSC, cryopreservation effects need to be assessed some time after, rather than immediately after thawing. MSCs cryopreserved with HES maintain their characteristic cell surface marker expression as well as the osteogenic, adipogenic and chondrogenic differentiation potential. HES alone does not provide sufficient cryoprotection for rat MSCs, but provides good cryoprotection in combination with DMSO, permitting the DMSO content to be reduced to 5%. There are indications that such a combination would seem useful not just for the clinical disadvantages of DMSO but also based on a tendency for reduced osteogenic differentiation capacity of rat MSC cryopreserved with high DMSO concentration. HES molecular weight appears to play only a minor role in its capacity to act as a cryopreservation solution for MSC. The use of a ‘straight freeze’ protocol is no less effective in maintaining post-thaw viability of MSC compared to controlled rate freezing methods. Conclusion A 5% DMSO / 5% HES solution cryopreservation solution using a ‘straight freeze’ approach can be recommended for rat MSC.

  5. The preventive effects of neural stem cells and mesenchymal stem cells intra-ventricular injection on brain stroke in rats

    Directory of Open Access Journals (Sweden)

    Seyed Mojtaba Hosseini

    2015-01-01

    Full Text Available Introduction: Stroke is one of the most important causes of disability in developed countries and, unfortunately, there is no effective treatment for this major problem of central nervous system (CNS; cell therapy may be helpful to recover this disease. In some conditions such as cardiac surgeries and neurosurgeries, there are some possibilities of happening brain stroke. Inflammation of CNS plays an important role in stroke pathogenesis, in addition, apoptosis and neural death could be the other reasons of poor neurological out come after stroke. In this study, we examined the preventive effects of the neural stem cells (NSCs and mesenchymal stem cells (MSCs intra-ventricular injected on stroke in rats. Aim: The aim of this study was to investigate the preventive effects of neural and MSCs for stroke in rats. Materials and Methods: The MSCs were isolated by flashing the femurs and tibias of the male rats with appropriate media. The NSCs were isolated from rat embryo ganglion eminence and they cultured NSCs media till the neurospheres formed. Both NSCs and MSCs were labeled with PKH26-GL. One day before stroke, the cells were injected into lateral ventricle stereotactically. Results: During following for 28 days, the neurological scores indicated that there are better recoveries in the groups received stem cells and they had less lesion volume in their brain measured by hematoxylin and eosin staining. Furthermore, the activities of caspase-3 were lower in the stem cell received groups than control group and the florescent microscopy images showed that the stem cells migrated to various zones of the brains. Conclusion: Both NSCs and MSCs are capable of protecting the CNS against ischemia and they may be good ways to prevent brain stroke consequences situations.

  6. Chemically Induced Breast Tumors in Rats Are Detectable in Early Stages by Contrast Enhanced Magnetic Resonance Imaging but Not by Changes in the Acute-Phase Reactants in Serum

    Directory of Open Access Journals (Sweden)

    Onn Haji Hashim

    2011-02-01

    Full Text Available The present study was undertaken to develop a rat model for monitoring the early development of breast cancer. Twelve female rats were divided into two groups of six rats that were either treated with N-methyl-N-nitrosourea to induce breast cancer or with bacterial lipopolysaccharide to induce inflammation. Serum samples taken from the rats prior to the treatment were used as controls. By the 14th week, presence of the tumor was detectable by contrast enhanced magnetic resonance imaging and confirmed by histopathology. When the serum proteins of the rats were examined by 2-dimensional electrophoresis (2-DE, no difference could be detected in the profiles of all proteins before and 18 weeks after administration of N-methyl-N-nitrosourea. However, higher expression of alpha-1B glycoprotein was detectable by 2-DE in serum samples of rats at the 18th week post-treatment with lipopolysaccharide.

  7. Cholangiocytes with Mesenchymal Features Contribute to Progressive Hepatic Fibrosis of the Polycystic Kidney Rat

    OpenAIRE

    Sato, Yasunori; Harada, Kenichi; Ozaki, Satoru; Furubo, Shinichi; Kizawa, Kazuo; Sanzen, Takahiro; Yasoshima, Mitsue; Ikeda, Hiroko; Sasaki, Motoko; Nakanuma, Yasuni

    2007-01-01

    The polycystic kidney (PCK) rat is an animal model of Caroli’s disease with congenital hepatic fibrosis, in which the mechanism of progressive hepatic fibrosis remains unknown. This study aimed to clarify the mechanism of hepatic fibrosis of the PCK rat from the viewpoint of the contribution of pathological cholangiocytes. In liver sections of the PCK rats, intrahepatic bile ducts were constituted by two different phenotypes: bile ducts lined by cuboidal-shaped and flat-shaped cholangiocytes....

  8. Adenovirus-Mediated Over-Expression of Nrf2 Within Mesenchymal Stem Cells (MSCs Protected Rats Against Acute Kidney Injury

    Directory of Open Access Journals (Sweden)

    Mohammad Mohammadzadeh-Vardin

    2015-06-01

    Full Text Available Purpose: Recent developments in the field of cell therapy have led to a renewed interest in treatment of acute kidney injury (AKI. However, the early death of transplanted mesenchymal stem cells (MSCs in stressful microenvironment of a recipient tissue is a major problem with this kind of treatment. The objective of this study was to determine whether overexpression of a cytoprotective factor, nuclear factor erythroid-2 related factor 2 (Nrf2, in MSCs could protect rats against AKI. Methods: The Nrf2 was overexpressed in MSCs by recombinant adenoviruses, and the MSCs were implanted to rats suffering from cisplatin-induced AKI. Results: The obtained results showed that transplantation with the engineered MSCs ameliorates cisplatin-induced AKI. Morphologic features of the investigated kidneys showed that transplantation with the MSCs in which Nrf2 had been overexpressed significantly improved the complications of AKI. Conclusion: These findings suggested that the engineered MSCs might be a good candidate to be further evaluated in clinical trials. However, detailed studies must be performed to investigate the possible carcinogenic effect of Nrf2 overexpression.

  9. Pre-conditioned mesenchymal stem cells ameliorate renal ischemic injury in rats by augmented survival and engraftment

    Directory of Open Access Journals (Sweden)

    Masoud Muhammad

    2012-12-01

    Full Text Available Abstract Background Ischemia is the major cause of acute kidney injury (AKI, associated with high mortality and morbidity. Mesenchymal stem cells (MSCs have multilineage differentiation potential and can be a potent therapeutic option for the cure of AKI. Methods MSCs were cultured in four groups SNAP (S-nitroso N-acetyl penicillamine, SNAP + Methylene Blue (MB, MB and a control for in vitro analysis. Cultured MSCs were pre-conditioned with either SNAP (100 μM or MB (1 μM or both for 6 hours. Renal ischemia was induced in four groups (as in in vitro study of rats by clamping the left renal padicle for 45 minutes and then different pre-conditioned stem cells were transplanted. Results We report that pre-conditioning of MSCs with SNAP enhances their proliferation, survival and engraftment in ischemic kidney. Rat MSCs pre-conditioned with SNAP decreased cell apoptosis and increased proliferation and cytoprotective genes’ expression in vitro. Our in vivo data showed enhanced survival and engraftment, proliferation, reduction in fibrosis, significant improvement in renal function and higher expression of pro-survival and pro-angiogenic factors in ischemic renal tissue in SNAP pre-conditioned group of animals. Cytoprotective effects of SNAP pre-conditioning were abrogated by MB, an inhibitor of nitric oxide synthase (NOS and guanylate cyclase. Conclusion The results of these studies demonstrate that SNAP pre-conditioning might be useful to enhance therapeutic potential of MSCs in attenuating renal ischemia reperfusion injury.

  10. Differentiation of rat adipose tissue-derived mesenchymal stem cells towards a nucleus pulposus-like phenotype in vitro

    Institute of Scientific and Technical Information of China (English)

    XIE Li-wei; FANG Huang; CHEN An-min; LI Feng

    2009-01-01

    Objective: To differentiate rat adipose tissue-derived mesenchymal stem cells (ADSCs) into cells with a nucleus pulposus-like phenotype in vitro,so as to lay a foundation for the cell-based transplantation therapy of degenerated intervertebral discs.Methods: Rat ADSCs were isolated only from the subcutaneous inguinal region and purified by limited dilution.ADSCs of the third passages were analyzed by fluorescence activated cell sorter (FACS) to detect the cell surface markers (Sca-1,CD44,CD45,CD11b).To induce ADSCs towards a nucleus pulposus-like phenotype,ADSCs were immobilized in 3-dimensional alginate hydrogels and cultured in an inducing medium containing transforming growth factor-betal (TGF-β1) under hypoxia (2% O2),while control groups under normoxia (21% O2) in alginate beads in medium with or without the presence of TGF-β1.Semiquantitative reverse transcription polymerase chain reaction (RT-PCR) was carded out to evaluate phenotypic and biosynthetic activities in the process of differentiation.Meanwhile,Alcian blue staining were used to detect the formation of sulfated glycosaminoglycans (GAGs) in the differentiated cells.Results: The purified ADSCs were fibroblast-like and proliferated rapidly in vitro.The flow cytometry showed that ADSCs were positive for Sea-1 and CD44,negative for CD45 and CD11b.The results of RT-PCR manifested that the gene expressions of Sox-9,aggrecan and collagen Ⅱ,which were chondrocyte specific,were upregulated in medium containing TGF-β1 under hypoxia (2% O2).Likewise,gene expression of HIF-la,which was characteristics of intervertebral discs,was also upregulated.Simultaneously,Alcian blue staining exhibited the formation of many GAGs.Conclusions: The approach in our experiment is a simple and effective way to acquire a large quantity of homogenous ADSCs.Rat ADSCs can be differentiated into nucleus pulposus-like cells.ADSCs may replace bone marrow mesenchymal stem cells as a new kind of seed cells in regeneration of

  11. The Cotransplantation of Olfactory Ensheathing Cells with Bone Marrow Mesenchymal Stem Cells Exerts Antiapoptotic Effects in Adult Rats after Spinal Cord Injury

    OpenAIRE

    Shifeng Wu; Guanqun Cui; Hua Shao; Zhongjun Du; Ng, Jack C.; Cheng Peng

    2015-01-01

    The mechanisms behind the repairing effects of the cotransplantation of olfactory ensheathing cells (OECs) with bone marrow mesenchymal stromal cells (BMSCs) have not been fully understood. Therefore, we investigated the effects of the cotransplantation of OECs with BMSCs on antiapoptotic effects in adult rats for which the models of SCI are induced. We examined the changes in body weight, histopathological changes, apoptosis, and the expressions of apoptosis-related proteins after 14 days an...

  12. New Therapy of Skin Repair Combining Adipose-Derived Mesenchymal Stem Cells with Sodium Carboxymethylcellulose Scaffold in a Pre-Clinical Rat Model

    OpenAIRE

    Cristiano Rodrigues; de Assis, Adriano M.; Moura, Dinara J.; Graziele Halmenschlager; Jenifer Saffi; Léder Leal Xavier; Marilda da Cruz Fernandes; Márcia Rosângela Wink

    2014-01-01

    Lesions with great loss of skin and extensive burns are usually treated with heterologous skin grafts, which may lead rejection. Cell therapy with mesenchymal stem cells is arising as a new proposal to accelerate the healing process. We tested a new therapy consisting of sodium carboxymethylcellulose (CMC) as a biomaterial, in combination with adipose-derived stem cells (ADSCs), to treat skin lesions in an in vivo rat model. This biomaterial did not affect membrane viability and induced a sma...

  13. Long Term Study of Protective Mechanisms of Human Adipose Derived Mesenchymal Stem Cells on Cisplatin Induced Kidney injury in Sprague-Dawley Rats

    OpenAIRE

    Elhusseini, Fatma M; Saad, Mohamed-Ahdy A.A; Anber, Nahla; Elghannam, Doaa; Sobh, Mohamed-Ahmed; ALSAYED, AZIZA; El-dusoky, Sara; SHEASHAA, HUSSEIN; Abdel-Ghaffar, Hassan; Sobh, Mohamed

    2016-01-01

    Background/Aims: Long-term evaluation of cisplatin induced nephrotoxicity and the probable renal protective activities of stem cells are lacking up until now. We evaluated the early and long-term role of human adipose derived mesenchymal stem cells (ADMSCs) in prevention or amelioration of cisplatin induced acute kidney injury (AKI) in Sprague-Dawley rats. For this, we determined the kidney tissue level of oxidative stress markers in conjugation with a renal histopathological scoring system o...

  14. Direct muscle delivery of GDNF with human mesenchymal stem cells improves motor neuron survival and function in a rat model of familial ALS

    OpenAIRE

    Suzuki, Masatoshi; McHugh, Jacalyn; Tork, Craig; Shelley, Brandon; Hayes, Antonio; Bellantuono, Ilaria; Aebischer, Patrick; Svendsen, Clive N.

    2008-01-01

    Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disease in which there is a progressive loss of motor neurons and their connections to muscle leading to paralysis. To maintain muscle connections in a rat model of familial ALS, we performed intramuscular transplantation with human mesenchymal stem cells (hMSC) as “Trojan horses” to deliver growth factors to the terminals of motor neurons as well as the skeletal muscles. hMSC engineered to secrete glial cell line derived neurot...

  15. Gadolinium and fluorescent bi-functionally labeling and in vitro MRI of rat bone marrow mesenchymal stem cells

    International Nuclear Information System (INIS)

    Objective: To determine the feasibility of magnetically labeling and tracking mesenchymal stem cells (MSCs) in vitro by using a gadolinium and fluorescent bi-functionally transfection agent of polyethylenimine. Methods: A gadolinium bifunctional transfection reagent complex was obtained after the linear polyethylenimine derivative (JetPEI-FluoR) was incubated with Gd-DTPA. Mesenchymal stem cells isolated from the bone marrows of SD rats were cultured and expanded. The mesenchymal stem cells were incubated with the bi-functional labeling agents. After labeling, the MSCs were examined with fluoroscope and electron microscope and the biological characters were detected including trypan blue exclusion test, MTT, and apoptosis detection. On a 1.5 T MR system, the labeled MSCs were examined with spin echo T1WI and T2WI and T1 measurement with mixed sequence. After labeling, the cells were cultured and undergone routine passage. Prior MR examinations were repeated for each passage of labeled cells. All data was statistically prolessed with SPSS for Windows. Results: Of 5 x 105 MSCs incubated with the bi-functional agents, 4.25 x 105 MSCs were successfully labeled, the percentage of labeled MSCs was 85% fluoroscopically. The high density electron particles of gadolinium observed electron microscopically existed around cellular apparatuses, especially around Golgi apparatus. In trypan blue exclusion test, the exclusion rate of labeled MSCs with incubation duration of 3,6,12,24 h was (96.55±2.90)%, (94.17± 2.56)%, (97.16±3.12)% and (94.23±2.67)%, respectively. The corresponding exclusion rate of unlabeled MSCs was (95.86±2.67)%, (92.04±2.21)%, (93.38±3.64)% and (92.12±2.53)%, respectively. There was no statistical difference of trypan blue exclusion rate between labeled cells and control unlabeled cells within 24 hours of incubation (F=4.523, P>0.05). In the proliferation test, the optical absorption value of labeled MSC with 2.5, 5.0, 10.0, 20.0, 30.0 and 40.0

  16. Propofol injection combined with bone marrow mesenchymal stem cell transplantation better improves electrophysiological function in the hindlimb of rats with spinal cord injury than monotherapy

    Institute of Scientific and Technical Information of China (English)

    Yue-xin Wang; Jing-jing Sun; Mei Zhang; Xiao-hua Hou; Jun Hong; Ya-jing Zhou; Zhi-yong Zhang

    2015-01-01

    The repair effects of bone marrow mesenchymal stem cell transplantation on nervous system damage are not satisfactory. Propofol has been shown to protect against spinal cord injury. Therefore, this study sought to explore the therapeutic effects of their combination on spinal cord injury. Rat models of spinal cord injury were established using the weight drop method. Rats were subjected to bone marrow mesenchymal stem cell transplantationvia tail vein injection and/or propofol injectionvia tail vein using an infusion pump. Four weeks after cell transplan-tation and/or propofol treatment, the cavity within the spinal cord was reduced. The numbers of PKH-26-positive cells and horseradish peroxidase-positive nerve ifbers apparently increased in the spinal cord. Latencies of somatosensory evoked potentials and motor evoked potentials in the hindlimb were noticeably shortened, amplitude was increased and hindlimb motor function was obviously improved. Moreover, the combined effects were better than cell transplantation or propofol injection alone. The above data suggest that the combination of propofol injection and bone marrow mesenchymal stem cell transplantation can effectively improve hindlimb electro-physiological function, promote the recovery of motor funtion, and play a neuroprotective role in spinal cord injury in rats.

  17. Propofol combined with bone marrow mesenchymal stem cell transplantation improves electrophysiological function in the hindlimb of rats with spinal cord injury better than monotherapy

    Directory of Open Access Journals (Sweden)

    Yue-xin Wang

    2015-01-01

    Full Text Available The repair effects of bone marrow mesenchymal stem cell transplantation on nervous system damage are not satisfactory. Propofol has been shown to protect against spinal cord injury. Therefore, this study sought to explore the therapeutic effects of their combination on spinal cord injury. Rat models of spinal cord injury were established using the weight drop method. Rats were subjected to bone marrow mesenchymal stem cell transplantation via tail vein injection and/or propofol injection via tail vein using an infusion pump. Four weeks after cell transplantation and/or propofol treatment, the cavity within the spinal cord was reduced. The numbers of PKH-26-positive cells and horseradish peroxidase-positive nerve fibers apparently increased in the spinal cord. Latencies of somatosensory evoked potentials and motor evoked potentials in the hindlimb were noticeably shortened, amplitude was increased and hindlimb motor function was obviously improved. Moreover, the combined effects were better than cell transplantation or propofol injection alone. The above data suggest that the combination of propofol injection and bone marrow mesenchymal stem cell transplantation can effectively improve hindlimb electrophysiological function, promote the recovery of motor funtion, and play a neuroprotective role in spinal cord injury in rats.

  18. Cryopreservation of Rat Bone Marrow Derived Mesenchymal Stem Cells by Two Conventional and Open-pulled Straw Vitrification Methods

    Directory of Open Access Journals (Sweden)

    Mohammad Hadi Bahadori

    2009-01-01

    Full Text Available Objective: Mesenchymal stem cells (MSCs are obtained from a variety of sources, mainlythe bone marrow. These cells have a great potential for clinical research, however they cannotstay alive for long periods in culture. The aim of this study is to determine whether vitrificationcan be a useful freezing method for the storage of MSCs.Materials and Methods: Mesenchymal stem cells were isolated from rat bone marrow basedon their capacity to adhere to plastic culture surfaces. MSCs were cryopreserved using boththe vitrification method and open-pulled straw (OPS vitrification and stored in liquid nitrogenwith ethylene glycol ficoll (EFS as a cryoprotectant for two months. The morphology andviability of thawed MSCs were evaluated by trypan blue staining. Furthermore, pre and postcryopreserved MSCs were induced to osteocyte and adipocyte with corresponding osteogenicand adipogenic medium.Results: After thawing, the viability rates were 81.33% ± 6.83 for the vitrification method and80.83% ± 6.4 for OPS vitrification, while the values in the pre-vitrification control group were88.16% ± 6.3 (Mean ± SD, n = 6. Post-cryopreserved cells from both the vitrification methodand OPS vitrification also had a similar cellular morphology and colony-formation that wasindistinguishable from non-vitrified fresh MSCs. In addition, the resuscitated cells cultured ininduction medium showed osteogenesis. Mineral production and deposition was detectableby alizarine red S staining. Moreover, by applying an adipogenic differentiation condition,both pre and post cryopreserved cells differentiated into adipocyte and lipid vacuole accumulationthat was stained by oil red O.Conclusion: Vitrification is a reliable and effective method for the cryopreservation of MSCs.

  19. Human mesenchymal stem cells modulate inflammatory cytokines after spinal cord injury in rat

    Czech Academy of Sciences Publication Activity Database

    Machová-Urdzíková, Lucia; Růžička, Jiří; LaBagnara, M.; Kárová, Kristýna; Kubinová, Šárka; Jiráková, Klára; Murali, R.; Syková, Eva; Jhanwar-Uniyal, M.; Jendelová, Pavla

    2014-01-01

    Roč. 15, č. 7 (2014), s. 11275-11293. E-ISSN 1422-0067 R&D Projects: GA ČR GP13-15031P; GA ČR(CZ) GA13-00939S; GA MŠk LH12024; GA MŠk EE2.3.30.0018; GA MŠk(CZ) ED1.1.00/02.0109 Grant ostatní: GAUK(CZ) 521712 Institutional support: RVO:68378041 Keywords : mesenchymal stem cells * spinal cord injury * inflammatory cytokines Subject RIV: FH - Neurology Impact factor: 2.862, year: 2014

  20. Comparison of articular cartilage repair with different hydrogel-human umbilical cord blood-derived mesenchymal stem cell composites in a rat model

    OpenAIRE

    Chung, Jun Young; Song, Minjung; Ha, Chul-Won; Kim, Jin-A; Lee, Choong-Hee; Park, Yong-Beom

    2014-01-01

    Introduction The present work was designed to explore the feasibility and efficacy of articular cartilage repair using composites of human umbilical cord blood derived mesenchymal stem cells (hUCB-MSCs) and four different hydrogels in a rat model. Methods Full-thickness articular cartilage defects were created at the trochlear groove of femur in both knees of rats. Composites of hUCB-MSCs and four different hydrogels (group A, 4% hyaluronic acid; group B, 3% alginate:30% pluronic (1:1, v/v); ...

  1. Biochemical and morphological changes in bone marrow mesenchymal stem cells induced by treatment of rats with p-Nonylphenol

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    Mohammad Hossein Abnosi

    2015-04-01

    Full Text Available Objective(s:In previous investigations, we have shown para-nonylphenol (p-NP caused significant reduction of proliferation and differentiation of rat bone marrow mesenchymal stem cells (MSCs in vitro. In this study, we first treat the rats with p-NP, then carried out the biochemical and morphological studies on MSCs. Materials and Methods: Proliferation property of cells was evaluated with the help of MTT assay, trypan blue, population doubling number, and colony forming assay. Differentiation property was evaluated with quantitative alizarin red assay, measurement of alkaline phosphatase (ALP activity as well as intracellular calcium content. In addition; morphological study, TUNEL test, activated caspase assay, and comet assay were performed to evaluate the mechanism of the cell death. Results: The results showed significant reduction in the colony-forming-ability and population-doubling-number of extracted cells when compared to control ones. In addition, it was revealed that the p-NP treatment of rats caused significant reduction in nuclear diameter, cytoplasm shrinkage, and induction of caspase-dependent-apoptosis. Also there was significant reduction in ALP activity, intracellular calcium content, and intracellular matrix following osteogenic differentiation. Conclusion: As MSCs are the cellular back up for bone remodeling and repair, we suggest more investigations to be conducted regarding the correlation between the increasing number of patients suffering from osteoporosis and p-NP toxicity. Also, we strongly recommend WHO and local health organization to prevent industries of using p-NP in formulation of industrial products which may cause changes in proliferation and differentiation properties of stem cells.

  2. CdSe/ZnS Quantum Dots-Labeled Mesenchymal Stem Cells for Targeted Fluorescence Imaging of Pancreas Tissues and Therapy of Type 1 Diabetic Rats

    Science.gov (United States)

    Liu, Haoqi; Tang, Wei; Li, Chao; Lv, Pinlei; Wang, Zheng; Liu, Yanlei; Zhang, Cunlei; Bao, Yi; Chen, Haiyan; Meng, Xiangying; Song, Yan; Xia, Xiaoling; Pan, Fei; Cui, Daxiang; Shi, Yongquan

    2015-06-01

    Mesenchymal stem cells (MSCs) have been used for therapy of type 1 diabetes mellitus. However, the in vivo distribution and therapeutic effects of transplanted MSCs are not clarified well. Herein, we reported that CdSe/ZnS quantum dots-labeled MSCs were prepared for targeted fluorescence imaging and therapy of pancreas tissues in rat models with type 1 diabetes. CdSe/ZnS quantum dots were synthesized, their biocompatibility was evaluated, and then, the appropriate concentration of quantum dots was selected to label MSCs. CdSe/ZnS quantum dots-labeled MSCs were injected into mouse models with type 1 diabetes via tail vessel and then were observed by using the Bruker In-Vivo F PRO system, and the blood glucose levels were monitored for 8 weeks. Results showed that prepared CdSe/ZnS quantum dots owned good biocompatibility. Significant differences existed in distribution of quantum dots-labeled MSCs between normal control rats and diabetic rats ( p pancreas of rats in the diabetes group, and was about 32 %, while that in the normal control group rats was about 18 %. The blood glucose levels were also monitored for 8 weeks after quantum dots-labeled MSC injection. Statistical differences existed between the blood glucose levels of the diabetic rat control group and MSC-injected diabetic rat group ( p pancreas tissues in diabetic rats, and significantly reduce the blood glucose levels in diabetic rats, and own potential application in therapy of diabetic patients in the near future.

  3. Hepatoblast and mesenchymal cell-specific gene-expression in fetal rat liver and in cultured fetal rat liver cells

    OpenAIRE

    Mansuroglu, Tümen; Dudás, József; Elmaouhoub, Abderrahim; Joza, Tobias; Ramadori, Giuliano

    2009-01-01

    The aim of this study was to determine whether passaged rat fetal liver cells are functional hepatoblasts. Hepatocyte/hepatoblast- and liver myofibroblast-gene-expressions were studied in adult and fetal rat liver tissues as well as in primary and passaged cultures of isolated rat fetal liver cells at both the mRNA and protein level. Desmin- and Alpha-Smooth Muscle Actin (SMA)-positive cells were located in the walls of liver vessels, whereas Desmin-positive/SMA-negative cells were distribute...

  4. Electrophysiological functional recovery in a rat model of spinal cord hemisection injury following bone marrow-derived mesenchymal stem cell transplantation under hypothermia

    Institute of Scientific and Technical Information of China (English)

    Dong Wang; Jianjun Zhang

    2012-01-01

    Following successful establishment of a rat model of spinal cord hemisection injury by resecting right spinal cord tissues, bone marrow stem cells were transplanted into the spinal cord lesions via the caudal vein while maintaining rectal temperature at 34 ± 0.5°C for 6 hours (mild hypothermia). Hematoxylin-eosin staining showed that astrocytes gathered around the injury site and formed scars at 4 weeks post-transplantation. Compared with rats transplanted with bone marrow stem cells under normal temperature, rats transplanted with bone marrow stem cells under hypothermia showed increased numbers of proliferating cells (bromodeoxyuridine-positive cells), better recovery of somatosensory-evoked and motor-evoked potentials, greater Basso, Beattie, and Bresnahan locomotor rating scores, and an increased degree of angle in the incline plate test. These findings suggested that hypothermia combined with bone marrow mesenchymal stem cells transplantation effectively promoted electrical conduction and nerve functional repair in a rat model of spinal cord hemisection injury.

  5. Use of hybrid chitosan membranes and human mesenchymal stem cells from the Wharton jelly of umbilical cord for promoting nerve regeneration in an axonotmesis rat model

    Institute of Scientific and Technical Information of China (English)

    Andrea G(a)rtner; Yuri Nakamura; Satoshi Hayakawa; Akiyoshi Osakah; Beatriz Porto; Ana Lúcia Luís; Artur SP Varej(a)o; Ana Colette Maurício; Tiago Pereira; Maria Jo(a)o Sim(o)es; Paulo AS Armada-da-Silva; Miguel L Fran(c)a; Rosa Sousa; Simone Bompasso; Stefania Raimondo; Yuki Shirosaki

    2012-01-01

    Many studies have been dedicated to the development of scaffolds for improving post-traumatic nerve regeneration.The goal of this study was to assess the effect on nerve regeneration,associating a hybrid chitosan membrane with non-differentiated human mesenchymal stem cells isolated from Wharton's jelly of umbilical cord,in peripheral nerve reconstruction after crush injury.Chromosome analysis on human mesenchymal stem cell line from Wharton's jelly was carried out and no structural alterations were found in metaphase.Chitosan membranes were previously tested in vitro,to assess their ability in supporting human mesenchymal stem cell survival,expansion,and differentiation.For the in vivo testing,Sasco Sprague adult rats were divided in 4 groups of 6 or 7 animals each:Group 1,sciatic axonotmesis injury without any other intervention (Group 1-Crush); Group 2,the axonotmesis lesion of 3 mm was infiltrated with a suspension of 1 250- 1 500 human mesenchymal stem cells (total volume of 50 μL) (Group 2-CrushCell); Group 3,axonotmesis lesion of 3 mm was enwrapped with a chitosan type Ⅲ membrane covered with a monolayer of non-differentiated human mesenchymal stem cells (Group 3-CrushChitlllCell) and Group 4,axonotmesis lesion of 3 mm was enwrapped with a chitosan type Ⅲ membrane (Group 4-CrushChitlll).Motor and sensory functional recovery was evaluated throughout a healing period of 12 weeks using sciatic functional index,static sciatic index,extensor postural thrust,and withdrawal reflex latency.Stereological analysis was carried out on regenerated nerve fibers.Results showed that infiltration of human mesenchymal stem cells,or the combination of chitosan membrane enwrapment and human mesenchymal stem cell enrichment after nerve crush injury provide a slight advantage to post-traumatic nerve regeneration.Results obtained with chitosan type Ⅲ membrane alone confirmed that they significantly improve post-traumatic axonal regrowth and may represent a very promising

  6. CdSe/ZnS Quantum Dots-Labeled Mesenchymal Stem Cells for Targeted Fluorescence Imaging of Pancreas Tissues and Therapy of Type 1 Diabetic Rats

    Science.gov (United States)

    Liu, Haoqi; Tang, Wei; Li, Chao; Lv, Pinlei; Wang, Zheng; Liu, Yanlei; Zhang, Cunlei; Bao, Yi; Chen, Haiyan; Meng, Xiangying; Song, Yan; Xia, Xiaoling; Pan, Fei; Cui, Daxiang; Shi, Yongquan

    2015-06-01

    Mesenchymal stem cells (MSCs) have been used for therapy of type 1 diabetes mellitus. However, the in vivo distribution and therapeutic effects of transplanted MSCs are not clarified well. Herein, we reported that CdSe/ZnS quantum dots-labeled MSCs were prepared for targeted fluorescence imaging and therapy of pancreas tissues in rat models with type 1 diabetes. CdSe/ZnS quantum dots were synthesized, their biocompatibility was evaluated, and then, the appropriate concentration of quantum dots was selected to label MSCs. CdSe/ZnS quantum dots-labeled MSCs were injected into mouse models with type 1 diabetes via tail vessel and then were observed by using the Bruker In-Vivo F PRO system, and the blood glucose levels were monitored for 8 weeks. Results showed that prepared CdSe/ZnS quantum dots owned good biocompatibility. Significant differences existed in distribution of quantum dots-labeled MSCs between normal control rats and diabetic rats ( p monitored for 8 weeks after quantum dots-labeled MSC injection. Statistical differences existed between the blood glucose levels of the diabetic rat control group and MSC-injected diabetic rat group ( p < 0.01), and the MSC-injected diabetic rat group displayed lower blood glucose levels. In conclusion, CdSe/ZnS-labeled MSCs can target in vivo pancreas tissues in diabetic rats, and significantly reduce the blood glucose levels in diabetic rats, and own potential application in therapy of diabetic patients in the near future.

  7. Effect of self-assembled peptide–mesenchymal stem cell complex on the progression of osteoarthritis in a rat model

    Directory of Open Access Journals (Sweden)

    Kim JE

    2014-05-01

    Full Text Available Ji Eun Kim,1 Sang Mok Lee,2 Soo Hyun Kim,1 Phil Tatman,3 Albert O Gee,4 Deok-Ho Kim,3,5 Kyung Eun Lee,6 Youngmee Jung,1 Sang Jun Kim21Biomaterials Research Center, Korea Institute of Science and Technology, Seoul, South Korea; 2Department of Physical and Rehabilitation Medicine, Samsung Medical Center, Seoul, South Korea; 3Department of Bioengineering, 4Department of Orthopaedics and Sports Medicine, 5Institute for Stem Cell and Regenerative Medicine and Center for Cardiovascular Biology, University of Washington, Seattle, WA, USA; 6Advanced Analysis Center, Korea Institute of Science and Technology, Seoul, South KoreaPurpose: To evaluate the efficacy of mesenchymal stem cells (MSCs encapsulated in self-assembled peptide (SAP hydrogels in a rat knee model for the prevention of osteoarthritis (OA progression.Materials and methods: Nanostructured KLD-12 SAPs were used as the injectable hydrogels. Thirty-three Sprague Dawley rats were used for the OA model. Ten rats were used for the evaluation of biotin-tagged SAP disappearance. Twenty-three rats were divided into four groups: MSC (n=6, SAP (n=6, SAP-MSC (n=6, and no treatment (n=5. MSCs, SAPs, and SAP-MSCs were injected into the knee joints 3 weeks postsurgery. Histologic examination, immunofluorescent staining, measurement of cytokine levels, and micro-computed tomography analysis were conducted 6 weeks after injections. Behavioral studies were done to establish baseline measurements before treatment, and repeated 3 and 6 weeks after treatment to measure the efficacy of SAP-MSCs.Results: Concentration of biotinylated SAP at week 1 was not significantly different from those at week 3 and week 6 (P=0.565. Bone mineral density was significantly lower in SAP-MSC groups than controls (P=0.002. Significant differences in terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling staining between the control group and all other groups were observed. Caspase-8, tissue inhibitor of

  8. Early life ethanol exposure causes long-lasting disturbances in rat mesenchymal stem cells via epigenetic modifications

    Energy Technology Data Exchange (ETDEWEB)

    Leu, Yu-Wei [Department of Life Science and Institute of Molecular Biology, National Chung Cheng University, Chia-Yi 621, Taiwan (China); Chu, Pei-Yi [Department of Pathology, Show Chwan Memorial Hospital, Changhua 500, Taiwan (China); Chen, Chien-Min [Division of Neurosurgery, Changhua Christian Hospital, Changhua 500, Taiwan (China); Yeh, Kun-Tu [Department of Pathology, Changhua Christian Hospital, Changhua 500, Taiwan (China); Liu, Yu Ming; Lee, Yen-Hui; Kuo, Shan-Tsu [Department of Life Science and Institute of Molecular Biology, National Chung Cheng University, Chia-Yi 621, Taiwan (China); Hsiao, Shu-Huei, E-mail: bioshh@ccu.edu.tw [Department of Life Science and Institute of Molecular Biology, National Chung Cheng University, Chia-Yi 621, Taiwan (China)

    2014-10-24

    Highlights: • Ethanol exposure alters proliferation and differentiation of MSCs. • Ethanol exposure suppresses osteogenesis and adipogenesis of MSCs. • H3K27me3-associated genes/pathways are affected in ethanol-exposed MSCs. • Expression of lineage-specific genes is dysregulated in ethanol-exposed MSCs. - Abstract: Fetal alcohol syndrome (FAS) is a birth defect due to maternal alcohol consumption during pregnancy. Because mesenchymal stem cells (MSCs) are the main somatic stem cells in adults and may contribute to tissue homeostasis and repair in adulthood, we investigated whether early life ethanol exposure affects MSCs and contributes to the propensity for disease onset in later life. Using a rodent model of FAS, we found that ethanol exposure (5.25 g/kg/day) from postnatal days 4 to 9 in rat pups (mimic of human third trimester) caused long-term anomalies in bone marrow-derived MSCs. MSCs isolated from ethanol-exposed animals were prone to neural induction but resistant to osteogenic and adipogenic inductions compared to their age-matched controls. The altered differentiation may contribute to the severe trabecular bone loss seen in ethanol-exposed animals at 3 months of age as well as overt growth retardation. Expression of alkaline phosphatase, osteocalcin, aP2, and PPARγ were substantially inhibited, but BDNF was up-regulated in MSCs isolated from ethanol-exposed 3 month-old animals. Several signaling pathways were distorted in ethanol-exposed MSCs via altered trimethylation at histone 3 lysine 27. These results demonstrate that early life ethanol exposure can have long-term impacts in rat MSCs by both genetic and epigenetic mechanisms.

  9. Mechano-growth factor induces migration of rat mesenchymal stem cells by altering its mechanical properties and activating ERK pathway

    International Nuclear Information System (INIS)

    Highlights: •MGF induced the migration of rat MSC in a concentration-dependent manner. •MGF enhanced the mechanical properties of rMSC in inducing its migration. •MGF activated the ERK 1/2 signaling pathway of rMSC in inducing its migration. •rMSC mechanics may synergy with ERK 1/2 pathway in MGF-induced rMSC migration. -- Abstract: Mechano-growth factor (MGF) generated by cells in response to mechanical stimulation has been identified as a mechano effector molecule, playing a key role in regulating mesenchymal stem cell (MSC) function, including proliferation and migration. However, the mechanism(s) underlying how MGF-induced MSC migration occurs is still unclear. In the present study, MGF motivated migration of rat MSCs (rMSCs) in a concentration-dependent manner and optimal concentration of MGF at 50 ng/mL (defined as MGF treatment in this paper) was demonstrated. Notably, enhancement of mechanical properties that is pertinent to cell migration, such as cell traction force and cell stiffness were found to respond to MGF treatment. Furthermore, MGF increased phosphorylation of extracellular signal-regulated kinase (ERK), ERK inhibitor (i.e., PD98059) suppressed ERK phosphorylation, and abolished MGF-induced rMSC migration were found, demonstrating that ERK is involved molecule for MGF-induced rMSC migration. These in vitro evidences of MGF-induced rMSC migration and its direct link to altering rMSC mechanics and activating the ERK pathway, uncover the underlying biomechanical and biological mechanisms of MGF-induced rMSC migration, which may help find MGF-based application of MSC in clinical therapeutics

  10. Early life ethanol exposure causes long-lasting disturbances in rat mesenchymal stem cells via epigenetic modifications

    International Nuclear Information System (INIS)

    Highlights: • Ethanol exposure alters proliferation and differentiation of MSCs. • Ethanol exposure suppresses osteogenesis and adipogenesis of MSCs. • H3K27me3-associated genes/pathways are affected in ethanol-exposed MSCs. • Expression of lineage-specific genes is dysregulated in ethanol-exposed MSCs. - Abstract: Fetal alcohol syndrome (FAS) is a birth defect due to maternal alcohol consumption during pregnancy. Because mesenchymal stem cells (MSCs) are the main somatic stem cells in adults and may contribute to tissue homeostasis and repair in adulthood, we investigated whether early life ethanol exposure affects MSCs and contributes to the propensity for disease onset in later life. Using a rodent model of FAS, we found that ethanol exposure (5.25 g/kg/day) from postnatal days 4 to 9 in rat pups (mimic of human third trimester) caused long-term anomalies in bone marrow-derived MSCs. MSCs isolated from ethanol-exposed animals were prone to neural induction but resistant to osteogenic and adipogenic inductions compared to their age-matched controls. The altered differentiation may contribute to the severe trabecular bone loss seen in ethanol-exposed animals at 3 months of age as well as overt growth retardation. Expression of alkaline phosphatase, osteocalcin, aP2, and PPARγ were substantially inhibited, but BDNF was up-regulated in MSCs isolated from ethanol-exposed 3 month-old animals. Several signaling pathways were distorted in ethanol-exposed MSCs via altered trimethylation at histone 3 lysine 27. These results demonstrate that early life ethanol exposure can have long-term impacts in rat MSCs by both genetic and epigenetic mechanisms

  11. Nano-scaled hydroxyapatite/silk fibroin sheets support osteogenic differentiation of rat bone marrow mesenchymal cells

    International Nuclear Information System (INIS)

    A novel biomaterial that was composed of nano-scaled sintered hydroxyapatite (HAp) and silk fibroin (SF) was fabricated. We cultured rat marrow mesenchymal cells (MMCs) on this biomaterial (nano-HAp/SF sheet), on bare SF sheets, and on tissue culture polystyrene (TCPS) dishes as controls, then evaluated cell adhesion, proliferation, and differentiation of the MMCs. After 1 h of culture, a large number of viable cells were observed on the nano-HAp/SF sheets in comparison to the controls. In addition, after 3 h of culture, the morphology of the cells on the nano-HAp/SF sheets was quite different from that on the SF sheets. MMCs extrude their cytoplasmic processes to nano-HAp particles and are well attached to the sheets. After 14 days of culture, under osteogenic conditions, the alkaline phosphatase (ALP) activity and bone-specific osteocalcin secretion of the cells on nano-HAp/SF sheets were higher than were those on the controls. These results indicated that the surface of the nano-HAp/SF sheets is covered with appropriate HAp crystal for MMC adhesion/proliferation and that the sheets effectively support the osteogenic differentiation of MMCs. Therefore, the nano-HAp/SF sheet is an effective biomaterial that is applicable in bone reconstruction surgery

  12. Micro-/Nano- sized hydroxyapatite directs differentiation of rat bone marrow derived mesenchymal stem cells towards an osteoblast lineage

    Science.gov (United States)

    Huang, Yan; Zhou, Gang; Zheng, Lisha; Liu, Haifeng; Niu, Xufeng; Fan, Yubo

    2012-03-01

    Regenerative medicine consisting of cells and materials provides a new way for the repair and regeneration of tissues and organs. Nano-biomaterials are highlighted due to their advantageous features compared with conventional micro-materials. The aim of this study is to investigate the effects of micro-/nano- sized hydroxyapatite (μ/n-HA) on the osteogenic differentiation of rat bone marrow derived mesenchymal stem cells (rBMSCs). μ/n-HA were prepared by a microwave synthesizer and precipitation method, respectively. Different sizes of μ/n-HA were characterized by IR, XRD, SEM, TEM and co-cultured with rBMSCs. It was shown that rBMSCs expressed higher levels of osteoblast-related markers by n-HA than μ-HA stimulation. The size of HA is an important factor for affecting the osteogenic differentiation of rBMSCs. This provides a new avenue for mechanistic studies of stem cell differentiation and a new approach to obtain more committed differentiated cells.

  13. Sustained and promoter dependent bone morphogenetic protein expression by rat mesenchymal stem cells after BMP-2 transgene electrotransfer

    Directory of Open Access Journals (Sweden)

    E Ferreira

    2012-07-01

    Full Text Available Transplantation of mesenchymal stem cells (MSCs with electrotransferred bone morphogenetic protein-2 (BMP-2 transgene is an attractive therapeutic modality for the treatment of large bone defects: it provides both stem cells with the ability to form bone and an effective bone inducer while avoiding viral gene transfer. The objective of the present study was to determine the influence of the promoter driving the human BMP-2 gene on the level and duration of BMP-2 expression after transgene electrotransfer into rat MSCs. Cytomegalovirus, elongation factor-1α, glyceraldehyde 3-phosphate dehydrogenase, and beta-actin promoters resulted in a BMP-2 secretion rate increase of 11-, 78-, 66- and 36-fold over respective controls, respectively. In contrast, the osteocalcin promoter had predictable weak activity in undifferentiated MSCs but induced the strongest BMP-2 secretion rates in osteoblastically-differentiated MSCs. Regardless of the promoter driving the transgene, a plateau of maximal BMP-2 secretion persisted for at least 21 d after the hBMP-2 gene electrotransfer. The present study demonstrates the feasibility of gene electrotransfer for efficient BMP-2 transgene delivery into MSCs and for a three-week sustained BMP-2 expression. It also provides the first in vitro evidence for a safe alternative to viral methods that permit efficient BMP-2 gene delivery and expression in MSCs but raise safety concerns that are critical when considering clinical applications.

  14. Adipose-derived mesenchymal stem cells from the sand rat: transforming growth factor beta and 3D co-culture with human disc cells stimulate proteoglycan and collagen type I rich extracellular matrix

    OpenAIRE

    Tapp, Hazel; Deepe, Ray; Ingram, Jane A; Kuremsky, Marshall; Hanley, Edward N; Gruber, Helen E.

    2008-01-01

    Introduction Adult mesenchymal stem cell therapy has a potential application in the biological treatment of disc degeneration. Our objectives were: to direct adipose-derived mesenchymal stem cells (AD-MSC) from the sand rat to produce a proteoglycan and collagen type I extracellular matrix (ECM) rich in known ECM components of the annulus fibrosis of disc; and to stimulate proteoglycan production by co-culture of human annulus cells with AD-MSC. Methods AD-MSC were isolated and characterised ...

  15. Potential of bone marrow mesenchymal stem cells in management of Alzheimer's disease in female rats.

    Science.gov (United States)

    Salem, Ahmed M; Ahmed, Hanaa H; Atta, Hazem M; Ghazy, Mohamed A; Aglan, Hadeer A

    2014-12-01

    Alzheimer's disease (AD) has been called the disease of the century with significant clinical and socioeconomic impacts. Pharmacological treatment has limited efficacy and only provides symptomatic relief without long-term cure. Accordingly, there is an urgent need to develop novel and effective medications for AD. Stem cell-based therapy is a promising approach to handling neurodegenerative diseases. Therefore, the current study aimed to explore the possible therapeutic role of single intravenous injection of bone marrow derived mesenchymal stem cells (BM-MSCs) after 4 months in management of AD in the experimental model. The work also extended to compare the therapeutic potential of BM-MSCs with 2 conventional therapies of AD; rivastigmine and cerebrolysin administered daily. BM-MSCs were able to home at the injured brains and produced significant increases in the number of positive cells for choline acetyltransferase (ChAT) and survivin expression, as well as selective AD indicator-1 (seladin-1) and nestin gene expression. Histopathological examination indicated that BM-MSCs could remove beta-amyloid plaques from hippocampus. Significant improvement in these biomarkers was similar to or better sometimes than the reference drugs, clearly showing the potential therapeutic role of BM-MSCs against AD through their anti-apoptotic, neurogenic and immunomodulatory properties. PMID:25044885

  16. Improving the osteogenesis of rat mesenchymal stem cells by chitosan-based-microRNA nanoparticles.

    Science.gov (United States)

    Wu, Guangsheng; Feng, Chao; Hui, Guangyan; Wang, Zhongshan; Tan, Jun; Luo, Lankun; Xue, Peng; Wang, Qintao; Chen, Xiguang

    2016-03-15

    MicroRNAs (miRNAs) play important roles in the osteogenic differentiation of stem cells. However, the application of miRNA in bone regeneration has been limited by its poor stability, low cellular uptake, and undesired immune response. In this study, chitosan (CS)/tripolyphosphate (TPP)/Hyaluronic Acid (HA) nanoparticles (CTH NPs) were prepared to deliver antimiR-138 to bone marrow mesenchymal stem cells (MSCs). The particle size, polydispersity index, and zeta potential of CTH NPs were related to the weight ratio of CS:TPP:HA. At optimum N/P ratio (20:1), the highest encapsulation efficiency was obtained. Both blank CTH NPs and CTH/antmiR-138 NPs exhibited no cytotoxicity to MSCs. A high transfection efficiency (nearly 70%) and significant enhancement of the osteogenesis of MSCs were observed. Above results demonstrated that CTH NPs was a potential candidate as an efficient non-viral miRNA vector to regulate the osteogenic differentiation of MSCs. PMID:26794737

  17. Simulated microgravity alters multipotential differentiation of rat mesenchymal stem cells in association with reduced telomerase activity

    Science.gov (United States)

    Sun, Lianwen; Gan, Bo; Fan, Yubo; Xie, Tian; Hu, Qinghua; Zhuang, Fengyuan

    Microgravity is one of the most important characteristics in space flight. Exposure to microgravity results in extensive physiological changes in humans. Bone loss is one of the changes with serious consequences; however, the mechanism retains unclear. As the origin of osteoprogenitors, mesenchymal stem cells (MSCs) may play an important role in it. After cultured under simulated microgravity (in a rotary cell culture system, RCCS), MSCs were stained using oil red O to identify adipocytes. The mRNA level of bone morphogenetic protein (BMP)-2 and peroxisome proliferators-activated receptor (PPAR) γ2 was determined by RT-PCR. Otherwise, MSCs were induced to osteogenic differentiation after microgravity culture, and then the activity of alkaline phosphatase (ALP) was determined by PNPP and the content of osteocalcin (OC) by ELISA. Furthermore, the telomerase activity in MSCs was measured by TRAP. The results showed that simulated microgravity inhibited osteoblastic differentiation and induced adipogenic differentiation accompanied by the change of gene expression of BMP-2 and PPARγ2 in MSCs. Meanwhile, the telomerase activity decreased significantly in MSCs under simulated microgravity. The reduced bone formation in space flight may partly be due to the altered potential differentiation of MSCs associated with telomerase activity which plays a key role in regulating the lifespan of cell proliferation and differentiation. Therefore, telomerase activation/replacement may act as a potential countermeasure for microgravity-induced bone loss.

  18. Mesenchymal Stem Cells Derived from Rat Epicardial Versus Epididymal Adipose Tissue

    OpenAIRE

    Mohamadreza Baghaban Eslaminejad; Soura Mardpour; Marzieh Ebrahimi

    2011-01-01

    Objective(s)Some investigation has indicated that adipose-derived stem cells possess different surface epitopes and differentiation potential according to the localization of fat pad from which the cells were derived. In the present study proliferation capacity and aging of such cells were explored.Materials and MethodsAdherent cells were isolated from the collagenase digests of adipose tissues excised from rat epicardial and epididymal regions and propagated with several subcultures. The cel...

  19. The effect of progesterone in the prevention of the chemically induced experimental colitis in rats Efeito da progesterona na prevenção de colite experimental induzida quimicamente em ratos

    OpenAIRE

    Oguzhan Karatepe; Merih Altiok; Muharrem Battal; Gulcin Kamali; Ahu Kemik; Timucin Aydin; Servet Karahan

    2012-01-01

    PURPOSE: To study the effects of progesterone on an experimental colitis model. METHODS: Wistar albino rats were treated subcutaneously with 2mg/kg once a day during seven days Colitis was induced by intrarectal administration of 5mg trinitrobenzene sulfonic acid (TNBS). Disease activities, macroscopic and microscopic scores were evaluated. To determine the response provoked by progesterone we measured Colonic malondialdehyde (MDA), TNF alfa, IL-6 and Nitric oxide (NO) levels in addition to t...

  20. The Effect of Aligned and Random Electrospun Fibrous Scaffolds on Rat Mesenchymal Stem Cell Proliferation

    Directory of Open Access Journals (Sweden)

    Zahra Zonoubi

    2012-01-01

    Full Text Available Objective: The development of combining mesenchymal stem cells (MSCs with surface modified three-dimensional (3D biomaterial scaffold provides a desirable alternative for replacement of damaged and diseased tissue. Nanofibrous scaffolds serve as suitable environment for cell attachment and proliferation due to their similarity to the physical dimension of the natural extracellular matrix. In this study the properties of plasma treated poly-C-caprolactone nanofiber scaffolds (p-PCL and unaltered PCL scaffolds were compared,and then p-PCL scaffolds were evaluated for MSC culture.Materials and Methods: Aligned and random PCL nanofibrus scaffolds were fabricatedby electrospining and their surface modified with O2 plasma treatment to enhanceMSC proliferation, adhesion and interaction. Chemical and mechanical characterizationswere carried out using scanning electron microscopy (SEM, water contact angle and tensile testing. Cell adhesion and morphology were evaluated using SEM 1 day after culture. Statistical analysis was carried out using one way analysis of variance(ANOVA.Results: The proliferation of MSCs were evaluated using 3-(4,5-Dimethylthiazol-2-yl-2,5-DiphenyltetrazoliumBromide(MTT assay on day 1, 3, and 5 after cell culture. Results showed that the numbers of cells that had grown on PCL nanofibrous scaffolds were significantly higher than those of control surfaces without nanofibers. Furthermore, the proliferation of MSCs on random nanofiber was significantly higher compared to that on aligned nanofiber.Conclusion: This study showed that while both aligned and random plasma treated PCL nanofibrous scaffold are more suitable substrates for MSC growth than tissue culture plates, random nanofiber best supported the proliferation of MSCs.

  1. Mesenchymal stem cells suppress lung inflammation and airway remodeling in chronic asthma rat model via PI3K/Akt signaling pathway

    OpenAIRE

    Lin, Hai-Yan; Xu, Lei; Xie, Shuan-shuan; Yu, Fei; Hu, Hai-Yang; Song, Xiao-lian; Wang, Chang-Hui

    2015-01-01

    Background: Mesenchymal stem cells (MSCs) came out to attract wide attention and had become one of the hotspots of most diseases’ research in decades. But at present, the mechanisms of how MSCs work on chronic asthma remain undefined. Our study aims at verifying whether MSCs play a role in preventing inflammation and airway remodeling via PI3K/AKT signaling pathway in the chronic asthma rats model. Methods: First, an ovalbumin (OVA)-induced asthma model was built. MSCs were administered to ov...

  2. Differentiation of Rat bone marrow Mesenchymal stem cells into Adipocytes and Cardiomyocytes after treatment with platelet lysate

    Science.gov (United States)

    Homayouni Moghadam, Farshad; Tayebi, Tahereh; Barzegar, Kazem

    2016-01-01

    Background: Mesenchymal stem cells (MSCs) are multipotential cells and their therapeutic potency is under intense investigation. Studying the effect of different induction factors on MSCs could increase our knowledge about the differentiation potency of these cells. One of the most important sources of these factors in mammalian body is platelet. Platelet lysate (PL) contains many growth factors and therefore, it can be used as a differentiation inducer. In the present study, the effect of PL on differentiation of rat bone marrow MSCs into cardiomyocytes was studied. Materials and Methods: To study the differentiation-inducing effect of PL, MSCs were treated with 2.5, 5 and 10% PL. Early results of this study showed that PL in high concentrations (10%) induces adipogenic differentiation of MSCs. Therefore, to evaluate differentiation to cardiomyocytes, MSCs were cultured in media containing lower levels of PL (2.5% and 5%) and then cardiomyogenic differentiation was induced by treatment with 5-azacytidine. Differentiation of MSCs was evaluated using direct observation of beating cells, immunostaining and real-time PCR techniques. Results: The results of qPCR showed that treatment with PL alone increased the expression of cardiac alpha actinin (CAA) being predictable by earlier observation of beating cells in PL-treated groups. The results of staining assays against cardiac alpha actinin also showed that there were stained cells in PL-treated groups. Conclusion: The results of the present study showed that PL is a powerful induction factor for differentiation of MSCs into different cell lines such as cardiomyocytes and adipocytes. PMID:27047647

  3. In Vitro Toxic Effects of Zinc Oxide Nanoparticles on Rat Adipose Tissue-Derived Mesenchymal Stem Cells

    Science.gov (United States)

    Orazizadeh, Mahmoud; Khodadadi, Ali; Bayati, Vahid; Saremy, Sadegh; Farasat, Maryam; Khorsandi, Layasadat

    2015-01-01

    Objective Zinc oxide nanoparticles (ZnO-NPs) are increasingly used in sunscreens, bio- sensors, food additives, pigments, manufacture of rubber products, and electronic materi- als. There are several studies about the effects of NPs on dermal fibroblast or keratino- cytes, but very little attention has been directed towards adipose-derived mesenchymal stem cells (ASCs). A previous study has revealed that ZnO-NPs restricted the migration capability of ASCs. However, the potential toxicity of these NPs on ASCs is not well un- derstood. This study intends to evaluate the effects of ZnO-NPs on subcutaneous ASCs. Materials and Methods In this experimental study, In order to assess toxicity, we ex- posed rat ASCs to ZnO-NPs at concentrations of 10, 50, and 100 µg/ml for 48 hours. Tox- icity was evaluated by cell morphology changes, cell viability assay, as well as apoptosis and necrosis detection. Results ZnO-NPs concentration dependently reduced the survival rates of ASCs as re- vealed by the trypan blue exclusion and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazo- lium-bromide (MTT) tests. ZnO-NPs, at concentrations of 10 and 50 µg/ml, induced a significant increase in apoptotic indices as shown by the annexin V test. The concentration of 10 µg/ml of ZnO-NPs was more toxic. Conclusion Lower concentrations of ZnO-NPs have toxic and apoptotic effects on subcutaneous ASCs. We recommend that ZnO-NPs be used with caution if there is a dermatological problem. PMID:26464812

  4. Adipose derived mesenchymal stem cells transplantation via portal vein improves microcirculation and ameliorates liver fibrosis induced by CCl4 in rats

    Directory of Open Access Journals (Sweden)

    Wang Yu

    2012-06-01

    Full Text Available Abstract Introduction Adipose derived mesenchymal stem cells (ADMSCs, carrying the similar characteristics to bone marrow mesenchymal stem cells, only much more abundant and easier to obtain, may be a promising treatment for liver fibrosis. We aim to investigate the therapeutic potential of ADMSCs transplantation in liver fibrosis caused by carbon tetrachloride (CCl4 in rats as well as its underlying mechanism, and to further explore the appropriate infusion pathway. Methods ADMSCs were isolated, cultured and identified. Placebo and ADMSCs were transplanted via portal vein and tail vein respectively into carbon tetrachloride (CCl4-induced liver fibrosis rats. Computed tomography (CT perfusion scan and microvessel counts were performed to measure the alteration of liver microcirculation after therapy. Liver function tests and histological findings were estimated. Results CT perfusion scan shown significant decrease of hepatic arterial perfusion index, significant increased portal vein perfusion, total liver perfusion in rats receiving ADMSCs from portal vein, and Factor VIII (FVIII immunohistochemical staining shown significant decrease of microvessels in rats receiving ADMSCs from portal vein, indicating microcirculation improvement in portal vein group. Vascular endothelial growth Factor (VEGF was significantly up-regulated in fibrosis models, and decreased after ADMSCs intraportal transplantation. A significant improvement of liver functional test and histological findings in portal vein group were observed. No significance was found in rats receiving ADMSCs from tail vein. Conclusions ADMSCs have a therapeutic effect against CCl4-mediated liver fibrosis. ADMSCs may benefit the fibrotic liver through alteration of microcirculation, evidenced by CT perfusion scan and down-regulation of VEGF. Intraportal transplantation is a better pathway than tail vein transplantation.

  5. Trianthema portulacastrum Linn. displays anti-inflammatory responses during chemically induced rat mammary tumorigenesis through simultaneous and differential regulation of NF-κB and Nrf2 signaling pathways.

    Science.gov (United States)

    Mandal, Animesh; Bishayee, Anupam

    2015-01-01

    Trianthema portulacastrum, a medicinal and dietary plant, has gained substantial importance due to its various pharmacological properties, including anti-inflammatory and anticarcinogenic activities. We have recently reported that a characterized T. portulacastrum extract (TPE) affords a considerable chemoprevention of 7,12-dimethylbenz(a)anthracene (DMBA)-induced rat mammary tumorigenesis though the underlying mechanisms are not completely understood. The objective of this study was to investigate anti-inflammatory mechanisms of TPE during DMBA mammary carcinogenesis in rats by monitoring cyclooxygenase-2 (COX-2), heat shock protein 90 (HSP90), nuclear factor-kappaB (NF-κB) and nuclear factor erythroid 2-related factor 2 (Nrf2). Mammary tumors were harvested from our previous study in which TPE (50-200 mg/kg) was found to inhibit mammary tumorigenesis in a dose-response manner. The expressions of intratumor COX-2, HSP90, NF-κB, inhibitory kappaB-alpha (IκBα) and Nrf2 were determined by immunohistochemistry. TPE downregulated the expression of COX-2 and HSP90, blocked the degradation of IκBα, hampered the translocation of NF-κB from cytosol to nucleus and upregulated the expression and nuclear translocation of Nrf2 during DMBA mammary carcinogenesis. These results in conjunction with our previous findings suggest that TPE prevents DMBA-induced breast neoplasia by anti-inflammatory mechanisms mediated through simultaneous and differential modulation of two interconnected molecular circuits, namely NF-κB and Nrf2 signaling pathways. PMID:25622256

  6. Lipopolysaccharide preconditioning enhances the efficacy of mesenchymal stem cells transplantation in a rat model of acute myocardial infarction

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    Wang Zhaojun

    2009-08-01

    Full Text Available Abstract Background Mesenchymal stem cells (MSCs-based regenerative therapy is currently regarded as an alternative approach to salvage the acute myocardial infarcted hearts. However, the efficiency of MSCs transplantation is limited by lower survival rate of engrafted MSCs. In previous study, we found that 1.0 μg/ml Lipopolysaccharide (LPS could protect MSCs against apoptosis induced by oxidative stress and meanwhile enhance the proliferation of MSCs. Therefore, in the present study, we firstly preconditioned MSCs with 1.0 μg/ml LPS, then transplanted MSCs into ischemic myocardium, and observed the survival and cardiac protective capacity of MSCs in a rat model of acute myocardial infarction. Furthermore, we tried to explore the underlying mechanisms and the role of Toll-like receptor-4 (TLR4 in the signal pathway of LPS-induced cardiac protection. Methods and results Acute myocardial infarction model was developed by left anterior descending coronary artery ligation. 60 rats were divided into 4 groups randomly and given an intramyocardial injection of one of the following treatments: 30 μl PBS (control group, 3 × 106 wild MSCs/30 μl (wMSCs group, 3 × 106 LPS-preconditioned wild MSCs/30 μl (LPS-wMSCs group, or 3 × 106 LPS-preconditioned TLR4 gene deleted MSCs/30 μl (LPS-tMSCs group. After 3 weeks, LPS-preconditioned wild MSCs transplantation ameliorated cardiac function and reduced fibrosis of infarcted myocardium. Vascular density was markedly increased in LPS-wMSCs group compared with other three groups. Survival rate of engrafted MSCs was elevated and apoptosis of myocardium was reduced in infarcted heart. Expression of vascular endothelial growth factor (VEGF and phospho-Akt was increased in the infarcted myocardium after transplantation of LPS-preconditioned MSCs. Conclusion LPS preconditioning enhanced survival of engrafted MSCs, stimulated expression of VEGF and activated PI3K/Akt pathway. LPS preconditioning before MSCs

  7. In Vitro Study of the Effect of Vitamin E on Viability, Morphological Changes and Induction of Osteogenic Differentiation in Adult Rat Bone Marrow Mesenchymal Stem Cells

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    M Soleimani Mehranjani

    2014-10-01

    Full Text Available Introduction: Vitamin E as a strong antioxidant plays an important role in inhibiting free radicals. Therefore, this study aimed to investigate the effect of vitamin E on the viability, morphology and osteogenic differentiation in bone marrow mesenchymal stem cells of an adult rat. Methods: The bone marrow mesenchymal stem cells were extracted using the flashing-out method. At the end of the third passage, cells were divided into groups of control and experimental. Experimental cells were treated withVitamin E (5,10,15,25,50,100,150μM for a period of 21 days in the osteogenic media containing 10% of fetal bovine serum. The cell viability, bone matrix mineralization, intercellular and extracellular calcium deposition, alkaline phosphatase activity, expression of genes and synthesis of proteins of osteopontin and osteocalcin as well as morphological changes of the cells were investigated. The study data was analyzed using one-way ANOVA and T-Test setting the significant P value at P<0.05. Results: Within vitamin- E treated cells, the mean viability, mean bone matrix mineralization, calcium deposition, alkaline phosphatase activity, expression and synthesis of osteopontin and osteocalcin of the mesenchymal stem cells treated with vitamin E significantly increased in a dose dependent manner. Also cytoplasm extensions were observed in the cells treated with vitamin E. Conclusion: Since vitamin E caused a significant increase in cell viability and osteogenic differentiation in the mesenchymal stem cells, therefore it can be utilized in order to increase cell differentiation and cell survival.

  8. [Therapeutical effect of growth-associated protein 43 (GAP43) gene-modified bone marrow mesenchymal stem cell transplantation on rat retinal degenerative diseases].

    Science.gov (United States)

    Yang, Juan; Xie, Maosong; Zheng, Weidong; Hu, Jianzhang; Qu, Qiang

    2016-08-01

    Objective To investigate the potential of the treatment of growth-associated protein 43 (GAP43) gene-modified bone marrow-derived mesenchymal stem cells (BMSCs) for retinitis pigmentosa (RP). Methods BMSCs were isolated and cultured by adherence method. By transfecting GAP43 gene into BMSCs via a lentivirus vector, we got GAP43 gene-modified BMSCs. Sixty-three Royal College of Surgeons (RCS) rats were randomly divided into three groups: experimental group, negative control group and blank control group. The experimental rats received subretinal injection of GAP43 gene-modified BMSCs. The negative control rats received subretinal injection of BMSCs. The control rats received subretinal injection of PBS. Thirty days after transplanting, the retinal thickness was detected by optical coherence tomography (OCT), and the expression of rhodopsin in RCS rat retinas was examined by Western blotting. Results Compared with the blank control group and the negative control group, 30 days after GAP43 gene-modified BMSC transplantation, the retinal thickness of the experimental group remarkably increased and the expression of rhodopsin significantly rose. Conclusion GAP43 gene-modified BMSC transplantation can increase survival photoreceptor cells and delay retinal degeneration. PMID:27412933

  9. Heat shock protein 60 affects behavioral improvement in a rat model of Parkinson's disease grafted with human umbilical cord mesenchymal stem cell-derived dopaminergic-like neurons.

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    Zhao, Can; Li, Hui; Zhao, Xian-Jing; Liu, Zheng-Xia; Zhou, Ping; Liu, Ying; Feng, Mei-Jiang

    2016-06-01

    Parkinson's disease (PD) is a neurodegenerative disorder that is caused by a loss of dopaminergic (DAergic) neurons in mesencephalic substantia nigra (SN). Human umbilical cord mesenchymal stem cells (hUC-MSCs) are capable of self-renewal and differentiation into multiple cell lineages, including DAergic neurons. Thus, hUC-MSCs could be a promising alternative to compensate for the loss of DAergic neurons in PD. In the current study, hUC-MSCs and hUC-MSCs-derived DAergic-like neurons were transplanted into the striatum and SN of a rat model of PD that is induced by 6-hydroxydopamine (6-OHDA). We evaluated their therapeutic effects on improving rotation behavior in the rat and on modulating the level of heat shock protein 60 (Hsp60) expression in the brain. After transplantation, an amelioration of rotation behavior was observed in rats that underwent cell grafting, and hUC-MSCs-derived DAergic-like neurons were superior to hUC-MSCs at inducing behavioral improvement. Western blot and immunohistochemistry analysis indicated significantly elevated levels of Hsp60 in cell-grafted rats compared to 6-OHDA-lesioned (PD) rats. These results demonstrate that hUC-MSCs-based cell transplantation is potential therapeutic treatment for PD, and hUC-MSCs-derived DAergic-like neurons appear to be favorable candidates for cell replacement therapy in PD. Finally, Hsp60 could be involved in a mechanism of behavioral recovery. PMID:26758268

  10. Transplantation of insulin-producing cells from umbilical cord mesenchymal stem cells for the treatment of streptozotocin-induced diabetic rats

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    Tsai Pei-Jiun

    2012-04-01

    Full Text Available Abstract Background Although diabetes mellitus (DM can be treated with islet transplantation, a scarcity of donors limits the utility of this technique. This study investigated whether human mesenchymal stem cells (MSCs from umbilical cord could be induced efficiently to differentiate into insulin-producing cells. Secondly, we evaluated the effect of portal vein transplantation of these differentiated cells in the treatment of streptozotocin-induced diabetes in rats. Methods MSCs from human umbilical cord were induced in three stages to differentiate into insulin-producing cells and evaluated by immunocytochemistry, reverse transcriptase, and real-time PCR, and ELISA. Differentiated cells were transplanted into the liver of diabetic rats using a Port-A catheter via the portal vein. Blood glucose levels were monitored weekly. Results Human nuclei and C-peptide were detected in the rat liver by immunohistochemistry. Pancreatic β-cell development-related genes were expressed in the differentiated cells. C-peptide release was increased after glucose challenge in vitro. Furthermore, after transplantation of differentiated cells into the diabetic rats, blood sugar level decreased. Insulin-producing cells containing human C-peptide and human nuclei were located in the liver. Conclusion Thus, a Port-A catheter can be used to transplant differentiated insulin-producing cells from human MSCs into the portal vein to alleviate hyperglycemia among diabetic rats.

  11. Neuronal-like differentiation of bone marrow-derived mesenchymal stem cells induced by striatal extracts from a rat model of Parkinson's disease

    Institute of Scientific and Technical Information of China (English)

    Xiaoling Qin; Wang Han; Zhigang Yu

    2012-01-01

    A rat model of Parkinson's disease was established by 6-hydroxydopamine injection into the medial forebrain bundle. Bone marrow-derived mesenchymal stem cells (BMSCs) were isolated from the femur and tibia, and were co-cultured with 10% and 60% lesioned or intact striatal extracts. The results showed that when exposed to lesioned striatal extracts, BMSCs developed bipolar or multi-polar morphologies, and there was an increase in the percentage of cells that expressed glial fibrillary acidic protein (GFAP), nestin and neuron-specific enolase (NSE). Moreover, the percentage of NSE-positive cells increased with increasing concentrations of lesioned striatal extracts. However, intact striatal extracts only increased the percentage of GFAP-positive cells. The findings suggest that striatal extracts from Parkinson's disease rats induce BMSCs to differentiate into neuronal-like cells in vitro.

  12. Effects of a hybrid micro/nanorod topography-modified titanium implant on adhesion and osteogenic differentiation in rat bone marrow mesenchymal stem cells

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    Zhang WJ

    2013-01-01

    Full Text Available Wenjie Zhang,1,2,* Zihui Li,3,* Qingfeng Huang,1 Ling Xu,1 Jinhua Li,3 Yuqin Jin,1,2 Guifang Wang,1,2 Xuanyong Liu,2 Xinquan Jiang11Department of Prosthodontics, 2Oral Bioengineering Laboratory, Shanghai Research Institute of Stomatology, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, 3State Key Laboratory of High Performance Ceramics and Superfine Microstructure, Shanghai Institute of Ceramics, Chinese Academy of Sciences, Shanghai, China *These authors contributed equally to this workBackground and methods: Various methods have been used to modify titanium implant surfaces with the aim of achieving better osseointegration. In this study, we fabricated a clustered nanorod structure on an acid-etched, microstructured titanium plate surface using hydrogen peroxide. We also evaluated biofunctionalization of the hybrid micro/nanorod topography on rat bone marrow mesenchymal stem cells. Scanning electron microscopy and x-ray diffraction were used to investigate the surface topography and phase composition of the modified titanium plate. Rat bone marrow mesenchymal stem cells were cultured and seeded on the plate. The adhesion ability of the cells was then assayed by cell counting at one, 4, and 24 hours after cell seeding, and expression of adhesion-related protein integrin β1 was detected by immunofluorescence. In addition, a polymerase chain reaction assay, alkaline phosphatase and Alizarin Red S staining assays, and osteopontin and osteocalcin immunofluorescence analyses were used to evaluate the osteogenic differentiation behavior of the cells.Results: The hybrid micro/nanoscale texture formed on the titanium surface enhanced the initial adhesion activity of the rat bone marrow mesenchymal stem cells. Importantly, the hierarchical structure promoted osteogenic differentiation of these cells.Conclusion: This study suggests that a hybrid micro/nanorod topography on a titanium surface fabricated by

  13. Chinese preparation Xuesaitong promotes the mobilization of bone marrow mesenchymal stem cells in rats with cerebral infarction

    OpenAIRE

    Bao-xia Zhang; Jin-sheng Zhang; Mei-mei Du; Xiao-ya Wang; Wei Li

    2016-01-01

    After cerebral ischemia, bone marrow mesenchymal stem cells are mobilized and travel from the bone marrow through peripheral circulation to the focal point of ischemia to initiate tissue regeneration. However, the number of bone marrow mesenchymal stem cells mobilized into peripheral circulation is not enough to exert therapeutic effects, and the method by which blood circulation is promoted to remove blood stasis influences stem cell homing. The main ingredient of Xuesaitong capsules is Pana...

  14. CXCL13 Promotes the Effect of Bone Marrow Mesenchymal Stem Cells (MSCs on Tendon-Bone Healing in Rats and in C3HIOT1/2 Cells

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    Feng Tian

    2015-01-01

    Full Text Available Objectives: Mesenchymal stem cells (MSCs are potential effective therapy for tissue repair and bone regeneration. In present study, the effects of CXC chemokine ligand-13 (CXCL13 were evaluated on tendon-bone healing of rats. Methods: Tendon bone healing of the rat model was established and biomechanical testing was performed at 2, 4, 8 weeks after surgery. Murine mesenchymal cell line (C3HIOT1/2 cells was cultured. The expression of miRNA-23a was detected by real-time PCR. The protein expression of ERK1/2, JNK and p38 was detected by western blotting. MiR-23a mimic and inhibitor were used to overexpress or silence the expression of miR-23a. Results: MSCs significantly elevated the levels of ultimate load to failure, stiffness and stress in specimens of rats, the effects of which were enhanced by CXCL13. The expression of miR-23a was down-regulated and the protein of ERK1/2 level was up-regulated by CXCL13 treatment in both in vivo and in vitro experiments. ERK1/2 expression was elevated by overexpression of miR-23a and reduced by miR-23a inhibitor. Conclusions: These findings revealed that CXCL13 promoted the tendon-bone healing in rats with MSCs treatment, and implied that the activation of ERK1/2 via miR-23a was involved in the process of MSCs treated bone regeneration.

  15. Retinal Electrophysiological Effects of Intravitreal Bone Marrow Derived Mesenchymal Stem Cells in Streptozotocin Induced Diabetic Rats

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    Akkoç, Tolga; Eraslan, Muhsin; Şahin, Özlem; Özkara, Selvinaz; Vardar Aker, Fugen; Subaşı, Cansu; Karaöz, Erdal; Akkoç, Tunç

    2016-01-01

    Diabetic retinopathy is the most common cause of legal blindness in developed countries at middle age adults. In this study diabetes was induced by streptozotocin (STZ) in male Wistar albino rats. After 3 months of diabetes, rights eye were injected intravitreally with green fluorescein protein (GFP) labelled bone marrow derived stem cells (BMSC) and left eyes with balanced salt solution (Sham). Animals were grouped as Baseline (n = 51), Diabetic (n = 45), Diabetic+BMSC (n = 45 eyes), Diabetic+Sham (n = 45 eyes), Healthy+BMSC (n = 6 eyes), Healthy+Sham (n = 6 eyes). Immunohistology analysis showed an increased retinal gliosis in the Diabetic group, compared to Baseline group, which was assessed with GFAP and vimentin expression. In the immunofluorescence analysis BMSC were observed to integrate mostly into the inner retina and expressing GFP. Diabetic group had prominently lower oscillatory potential wave amplitudes than the Baseline group. Three weeks after intravitreal injection Diabetic+BMSC group had significantly better amplitudes than the Diabetic+Sham group. Taken together intravitreal BMSC were thought to improve visual function. PMID:27300133

  16. Combined Bone Mesenchymal Stem Cell and Olfactory Ensheathing Cell Transplantation Promotes Neural Repair Associated With CNTF Expression in Traumatic Brain-Injured Rats.

    Science.gov (United States)

    Fu, Xue-Mei; Liu, Su-Juan; Dan, Qi-Qin; Wang, Yan-Ping; Lin, Na; Lv, Long-Yun; Zou, Yu; Liu, Su; Zhou, Xue; Wang, Ting-Hua

    2015-01-01

    This study examined the role of bone mesenchymal stem cell (BMSC) and olfactory ensheathing cell (OEC) cografting on neural function and underlying molecular mechanisms in acute stage of traumatic brain injury (TBI) rats. Eighty Sprague-Dawley (SD) female rats were randomly divided into five groups (n = 16 per category): sham operated group (Sham), weight-drop-induced TBI group (TBI), BMSC transplantation group (BMSC), OEC transplantation group (OEC), and cotransplantation group (CO). Eight rats were randomly selected from each group for behavioral and morphological assessment. Another category (n = 8 rats) was employed in the genetic expression detection. BMSCs were isolated from GFP mice and identified by CD44 antibody. OECs were isolated from the SD rats, identified by P75 antibody and labeled by Hoechst 33342. They were then transplanted into the surrounding tissue of the epicenter of TBI rats. The result of neurological severity scores revealed that BMSC or OEC transplantation alone and BMSC and OEC cografting significantly ameliorated the neurological deficits of TBI rats. Quantitative immunohistochemical analysis showed that graft-recipient animals possessed dramatically more neurons and regenerated axons and smaller amounts of astrocytes than controls 14 days posttransplantation (p < 0.05). However, the expressional level of ciliary neurotrophic factor significantly decreased in the cografting group as determined by RT-PCR (p < 0.05), and the Janus kinase/signal transducer and activator of transcription pathway was significantly activated at 7 days after cell transplantation (p < 0.05). This study is the first to report the role of cotransplantation of BMSCs and OECs in the therapy of TBI and explore its potential molecular mechanisms, therefore providing the important morphological and molecular biological evidence for the clinical application of BMSC and/or OEC transplantation in TBI. PMID:24612678

  17. Promoting effect of small molecules in cardiomyogenic and neurogenic differentiation of rat bone marrow-derived mesenchymal stem cells

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    Khanabdali R

    2015-12-01

    Full Text Available Ramin Khanabdali,1 Anbarieh Saadat,1 Maizatul Fazilah,1 Khairul Fidaa’ Khairul Bazli,1 Rida-e-Maria Qazi,2 Ramla Sana Khalid,2 Durriyyah Sharifah Hasan Adli,1 Soheil Zorofchian Moghadamtousi,1 Nadia Naeem,2 Irfan Khan,2 Asmat Salim,2 ShamsulAzlin Ahmad Shamsuddin,1 Gokula Mohan1 1Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia; 2Dr Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan Abstract: Small molecules, growth factors, and cytokines have been used to induce differentiation of stem cells into different lineages. Similarly, demethylating agents can trigger differentiation in adult stem cells. Here, we investigated the in vitro differentiation of rat bone marrow mesenchymal stem cells (MSCs into cardiomyocytes by a demethylating agent, zebularine, as well as neuronal-like cells by β-mercaptoethanol in a growth factor or cytokines-free media. Isolated bone marrow-derived MSCs cultured in Dulbecco’s Modified Eagle’s Medium exhibited a fibroblast-like morphology. These cells expressed positive markers for CD29, CD44, and CD117 and were negative for CD34 and CD45. After treatment with 1 µM zebularine for 24 hours, the MSCs formed myotube-like structures after 10 days in culture. Expression of cardiac-specific genes showed that treated MSCs expressed significantly higher levels of cardiac troponin-T, Nkx2.5, and GATA-4 compared with untreated cells. Immunocytochemical analysis showed that differentiated cells also expressed cardiac proteins, GATA-4, Nkx 2.5, and cardiac troponin-T. For neuronal differentiation, MSCs were treated with 1 and 10 mM β-mercaptoethanol overnight for 3 hours in complete and serum-free Dulbecco’s Modified Eagle’s Medium, respectively. Following overnight treatment, neuron-like cells with axonal and dendritic-like projections originating from the

  18. Effects of transplantation with bone marrow-derived mesenchymal stem cells modified by Survivin on experimental stroke in rats

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    Cheng Ronghua

    2011-07-01

    Full Text Available Abstract Background This study was performed to determine whether injury induced by cerebral ischemia could be further improved by transplantation with bone marrow-derived mesenchymal stem cells (MSCs modified by Survivin (SVV. Methods MSCs derived from bone marrow of male Sprague-Dawley rats were infected by the self-inactive lentiviral vector GCFU carrying green fluorescent protein (GFP gene and SVV recombinant vector (GCFU-SVV. In vitro, vascular endothelial growth factor (VEGF and basic fibroblast growth factor (bFGF were detected in infected MSCs supernatants under hypoxic conditions by ELSIA. In vivo, experiments consisted of three groups, one receiving intravenous injection of 500 μl of phosphate-buffered saline (PBS without cells (control group and two groups administered the same volume solution with either three million GFP-MSCs (group GFP or SVV/GFP-MSCs (group SVV. All animals were submitted to 2-hour middle cerebral artery occlusion (MCAO and then reperfusion. Differentiation and survival of the transplanted MSCs were determined by confocal microscope. Western blot was used to detect the expression of VEGF and bFGF in ischemic tissue. A 2,3,5-triphenyltetrazolium chloride (TTC staining was used to assess the infarct volume. Evaluation of neurological function was performed using a modified Neurological Severity Score (mNSS. Results In vitro, modification with SVV further increased secretion of VEGF and bFGF under hypoxic condition. In vivo, only very few transplantated cells co-expressed GFP and NeuN. The survival transplanted cells in the group SVV was 1.3-fold at 4 days after transplantation and 3.4-fold higher at 14 days after transplantation, respectively, when compared with group GFP. Expression of VEGF and bFGF in the ischemic tissue were further up-regulated by modification with SVV. Moreover, modification with SVV further reduced the cerebral infarct volume by 5.2% at 4 days after stroke and improved post

  19. Evaluation of trans-1-amino-3-18F-fluorocyclobutanecarboxylic acid accumulation in low-grade glioma in chemically induced rat models: PET and autoradiography compared with morphological images and histopathological findings

    International Nuclear Information System (INIS)

    Introduction: Magnetic resonance imaging (MRI) can have a problem to delineate diffuse gliomas with an intact blood–brain barrier (BBB) especially when a marked peritumoral edema is present. We evaluated the potential of trans-1-amino-3-18F-fluorocyclobutanecarboxylic acid (anti-18F-FACBC) positron emission tomography (PET) to delineate the extent of diffuse gliomas by comparing PET findings with autoradiography, in vivo and ex vivo MRI, and histopathology findings. Methods: Dynamic PET was performed in rats with N-ethyl-N-nitrosourea-induced glioma for 60 min after anti-18F-FACBC injection. Contrast-enhanced MRI was performed before or after PET. The PET images were fused with in vivo and ex vivo MR images, and histopathological images for direct comparisons. Autoradiograms were compared with the results of Evans Blue (EB) extravasation (to assess BBB integrity) and hematoxylin-eosin staining. Results: Histopathological examination, including EB extravasation assessment, and enhanced T1-weighted MRI identified several diffuse gliomas with slight BBB disruption, similar to low-grade human gliomas. Anti-18F-FACBC uptake was specific and high in the gliomas, irrespective of BBB integrity. Higher anti-18F-FACBC uptake corresponded to areas of T2 hyperintensity, independent of gadolinium enhancement. Ex vivo autoradiography also showed high anti-18F-FACBC accumulation in tumors lacking EB extravasation and a correlation between anti-18F-FACBC accumulation and tumor cell density, but not EB extravasation. Conclusions: Anti-18F-FACBC-PET allowed visualization of gliomas irrespective of BBB integrity. The tumor-to-normal uptake ratio of anti-18F-FACBC generally correlated with the relative cell density. Anti-18F-FACBC PET combined with MRI shows promise for preoperative glioma delineation. Advances in knowledge: Radiopharmaceuticals that cross the BBB, such as anti-18F-FACBC, are taken up by low-grade gliomas with equivocal MRI findings due to an intact BBB

  20. The effect of progesterone in the prevention of the chemically induced experimental colitis in rats Efeito da progesterona na prevenção de colite experimental induzida quimicamente em ratos

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    Oguzhan Karatepe

    2012-01-01

    Full Text Available PURPOSE: To study the effects of progesterone on an experimental colitis model. METHODS: Wistar albino rats were treated subcutaneously with 2mg/kg once a day during seven days Colitis was induced by intrarectal administration of 5mg trinitrobenzene sulfonic acid (TNBS. Disease activities, macroscopic and microscopic scores were evaluated. To determine the response provoked by progesterone we measured Colonic malondialdehyde (MDA, TNF alfa, IL-6 and Nitric oxide (NO levels in addition to the MPO (Myeloperoxidase and caspase-3 activities. RESULTS: Progesterone ameliorated significantly the macroscopic and microscopic scores. TNBS-induced colitis significantly increased the colonic MDA levels and caspase-3 activities in group 2 in comparison to the control group. The results of the study revealed a decline in MDA, NO, IL6 and TNF-α levels in the colon tissue and in blood due to progesterone therapy in group 3 when compared to the group 2, a significant improvement. Progesterone treatment was associated with decreased MDA, MPO, TNF alfa and caspase-3 activity. CONCLUSION: Progesterone therapy decreased oxidative damage in the colonic mucosa.OBJETIVO: Investigar os efeitos da progesterona em um modelo de colite experimental. MÉTODOS: Ratos albinos Wistar foram tratados subcutaneamente com 2mg/kg por dia durante sete dias. A colite foi induzida por administração intrarretal de 5mg ácido sulfônico trinitrobenzeno (TNBS. Foram avaliadas as atividades da doença, escores macroscópicos e microscópicos Para determinar a resposta provocada pela progesterona foi medida no cólon os níveis de malondialdeído (MDA, TNF alfa, IL-6 e óxido nítrico (NO, além da atividade da MPO (Myeloperoxidase e caspase-3. RESULTADOS: A progesterone melhorou significantemente os escores macroscópicos e microscópicos. A colite induzida pelo TNBS significantemente aumentou os níveis colônicos de MDA e a atividade da caspase-3 no grupo 2 em comparação com o grupo

  1. Peripheral nerve reconstruction with epsilon-caprolactone conduits seeded with vasoactive intestinal peptide gene-transfected mesenchymal stem cells in a rat model

    Science.gov (United States)

    Hernández-Cortés, P.; Toledo-Romero, M. A.; Delgado, M.; Sánchez-González, C. E.; Martin, F.; Galindo-Moreno, P.; O'Valle, F.

    2014-08-01

    Objective. Attempts have been made to improve nerve conduits in peripheral nerve reconstruction. We investigated the potential therapeutic effect of a vasoactive intestinal peptide (VIP), a neuropeptide with neuroprotective, trophic and developmental regulatory actions, in peripheral nerve regeneration in a severe model of nerve injury that was repaired with nerve conduits. Approach. The sciatic nerve of each male Wistar rat was transected unilaterally at 10 mm and then repaired with Dl-lactic-ɛ-caprolactone conduits. The rats were treated locally with saline, with the VIP, with adipose-derived mesenchymal stem cells (ASCs) or with ASCs that were transduced with the VIP-expressing lentivirus. The rats with the transected nerve, with no repairs, were used as untreated controls. At 12 weeks post-surgery, we assessed their limb function by measuring the ankle stance angle and the percentage of their muscle mass reduction, and we evaluated the histopathology, immunohistochemistry and morphometry of the myelinated fibers. Main results. The rats that received a single injection of VIP-expressing ASCs showed a significant functional recovery in the ankle stance angle (p = 0.049) and a higher number of myelinated fibers in the middle and distal segments of the operated nerve versus the other groups (p = 0.046). Significance. These results suggest that utilization of a cellular substrate, plus a VIP source, is a promising method for enhancing nerve regeneration using Dl-lactic-ɛ-caprolactone conduits and that this method represents a potential useful clinical approach to repairing peripheral nerve damage.

  2. Cell multiplication, apoptosis and p-Akt protein expression of bone mesenchymal stem cells of rat under hypoxia environment

    Institute of Scientific and Technical Information of China (English)

    Hongliang Kong; Ningning Liu; Xin Huo; Bo Wang; Haipeng Zhang; Mingyu Gao; Guoxian Qi

    2007-01-01

    Objective :To elucidate whether cell multiplication, apoptosis, glucose intake and p-Akt protein expression of bone Mesenchymal Stem Cells(MSCs) of rats is influenced by a hypoxic environment ex vivo. Methods:Passage 3 of bone marrow MSCs taken from Wistar rats, were cultured in a culturing chamber with 94%N2,1%O2, 5%CO2 at 37℃. At different hypoxia time points, 0,0.5,1,4 and 8 h, glucose uptake was assayed by using radiation isotope 3H-G, Apoptotic Rate(AR) and dead rate(DR) were analyzed by flow cytometry(FCM) after Annexin V/PI staining, cell multiplication(by MTT methods) and p-Akt protein by immunocytochemistry and western blot. Results:Assay for CD29+,CD44+,CD71+,CD34-, Tn T+(after 5-azacytidine agent inducing) and ALP+(after bone differentiation agent inducing) suggested these bone-derived cells were MSCs. The 3H-G intaking ratio (CPM/flask value:157 ± 11,110 ± 11,107 ± 13,103 ± 10,100 ± 9 and 98 ± 10) of MSCs at different hypoxia time points, significantly decreased compared to that of normoxia(P < 0.01) and tended to descend slowly with hypoxia time duration, for which there was no statistical significance(P > 0.05). The AR(0.09 ± 2.03%,12.9 ± 1.72%,13.7 ± 2.26%,13.8 ± 3.01% ,14.1 ± 2.78% and 14.7 ±4.01% at 0,0.5,1,4 and 8 h,respectively,P < 0.01) and DR (0.04 ± 1.79% ,0.93 ± 1.85% ,3.11 ± 2.14%,4.09 ± 2.36% ,4.72 ±2.05% and 4.91 ± 3.72% at 0,0.5,1,4 and 8 h, respectively, P < 0.05) at different hypoxia time points significantly increased compared to those time in normoxia; The AR further went up with time (P < 0.05), however there was no statistical significance(P > 0.05) for the DR. Optical absorption value of MTT methods at different hypoxia time points significantly decreased compared to those with a corresponding normoxia time (P < 0.01 ) and degraded with time (in an hypoxic environment -P < 0.01 ).IOD of p-Akt protein of MSCs at different hypoxia time points significantly increased (0.367 ± 0.031,0.556 ± 0

  3. Early distribution of intravenously injected mesenchymal stem cells in rats with acute brain trauma evaluated by 99mTc-HMPAO labeling

    International Nuclear Information System (INIS)

    Introduction: Stem cell tracking is essential for evaluation of its migration, transplantation and therapeutic response. The aim of this study was to evaluate early distribution of intravenously transplanted rat bone marrow mesenchymal stem cells (BMSCs) in rats with acute cerebral trauma by labeling with 99mTc-hexamethylpropyleneamine oxime (99mTc-HMPAO). Methods: 99mTc-HMPAO-labeled BMSCs were injected intravenously to trauma rats (n=14) and sham-operated controls (n=13). Gamma camera images were acquired at 4 h after injection, and then organs were removed for gamma counting. Confocal microscope was used to confirm the migration of 99mTc-BMSCs by co-labeling with PKH26. Cytometric analysis was performed to evaluate apoptotic or necrotic change until the seventh day after labeling. Results: 99mTc-BMSCs were distributed mostly to lungs, liver and spleen at 4 h, and uptake of these organs was not significantly different between traumatic rats and controls. Meanwhile, the cerebral uptake of 99mTc-BMSCs was significantly higher in the traumatic rats than in controls (0.40% vs. 0.20%; P=.0002). Additionally, 99mTc-BMSCs' uptake of traumatic hemisphere was significantly higher than that of contralateral ones (0.27% vs. 0.13%; P=.0001) in traumatic rats. Regardless of radiolabeling, BMSCs migrated to traumatic regions, but not to nontraumatic hemispheres. However, gamma camera failed to demonstrate 99mTc-BMSCs in traumatic hemispheres. No significant apoptotic or necrotic change was observed until 7 days after radiolabeling. Conclusions: Early distribution of BMSCs in traumatic brain disease could be monitored by 99mTc-labeling, which does not induce cellular death. However, our data showed that the amount of migrated 99mTc-BMSCs was not enough to be demonstrated by clinical gamma camera.

  4. Gender difference in the neuroprotective effect of rat bone marrow mesenchymal cells against hypoxia-induced apoptosis of retinal ganglion cells

    OpenAIRE

    Jing Yuan; Jian-xiong Yu

    2016-01-01

    Bone marrow mesenchymal stem cells can reduce retinal ganglion cell death and effectively prevent vision loss. Previously, we found that during differentiation, female rhesus monkey bone marrow mesenchymal stem cells acquire a higher neurogenic potential compared with male rhesus monkey bone marrow mesenchymal stem cells. This suggests that female bone marrow mesenchymal stem cells have a stronger neuroprotective effect than male bone marrow mesenchymal stem cells. Here, we first isolated and...

  5. Deletion of a conserved regulatory element required for Hmx1 expression in craniofacial mesenchyme in the dumbo rat: a newly identified cause of congenital ear malformation

    Directory of Open Access Journals (Sweden)

    Lely A. Quina

    2012-11-01

    Hmx1 is a homeodomain transcription factor expressed in the developing eye, peripheral ganglia, and branchial arches of avian and mammalian embryos. Recent studies have identified a loss-of-function allele at the HMX1 locus as the causative mutation in the oculo-auricular syndrome (OAS in humans, characterized by ear and eye malformations. The mouse dumbo (dmbo mutation, with similar effects on ear and eye development, also results from a loss-of-function mutation in the Hmx1 gene. A recessive dmbo mutation causing ear malformation in rats has been mapped to the chromosomal region containing the Hmx1 gene, but the nature of the causative allele is unknown. Here we show that dumbo rats and mice exhibit similar neonatal ear and eye phenotypes. In midgestation embryos, dumbo rats show a specific loss of Hmx1 expression in neural-crest-derived craniofacial mesenchyme (CM, whereas Hmx1 is expressed normally in retinal progenitors, sensory ganglia and in CM, which is derived from mesoderm. High-throughput resequencing of 1 Mb of rat chromosome 14 from dmbo/dmbo rats, encompassing the Hmx1 locus, reveals numerous divergences from the rat genomic reference sequence, but no coding changes in Hmx1. Fine genetic mapping narrows the dmbo critical region to an interval of ∼410 kb immediately downstream of the Hmx1 transcription unit. Further sequence analysis of this region reveals a 5777-bp deletion located ∼80 kb downstream in dmbo/dmbo rats that is not apparent in 137 other rat strains. The dmbo deletion region contains a highly conserved domain of ∼500 bp, which is a candidate distal enhancer and which exhibits a similar relationship to Hmx genes in all vertebrate species for which data are available. We conclude that the rat dumbo phenotype is likely to result from loss of function of an ultraconserved enhancer specifically regulating Hmx1 expression in neural-crest-derived CM. Dysregulation of Hmx1 expression is thus a candidate mechanism for congenital ear

  6. Ameliorative effects of human adipose tissue-derived mesenchymal stem cells on myelin basic protein-induced experimental autoimmune encephalomyelitis in Lewis rats

    Institute of Scientific and Technical Information of China (English)

    Myung-Soon Ko; Hyeong-geun Park; Young-Min Yun; Jeong Chan Ra; Taekyun Shin; Kyoung-Kap Lee

    2011-01-01

    Mesenchymal stem cells have been previously shown to exert an immunomodulatory function. The present study sought to investigate the effects of multipotential human adipose tissue-derived mesenchymal stem cells (hAdMSCs) on disease progression and cytokine expression in Lewis rats with experimental autoimmune encephalomyelitis (EAE) induced by myelin basic protein. The duration of EAE paralysis in the group treated on day 7 postimmunization with 5 × 106 hAdMSCs was significantly reduced compared with the vehicle-treated controls and the 1 × 106 hAdMSC- treated group. The duration of EAE paralysis in the groups treated with 5 × 106 hAdMSCs on both day 1 and day 7 postimmunization was significantly reduced compared with the vehicle-treated controls and the groups treated with 5 × 106 hAdMSCs on both day 7 and day 10 postimmunization. The mRNA expression of interleukin-10 and indoleamine 2, 3-dioxygenase was significantly decreased in the hAdMSC-treated group compared with the vehicle-treated group. These findings suggest that the ameliorative effects of hAdMSCs on EAE symptoms operate in a dose- and time-dependent manner and can be mediated in part by the ample production of anti-inflammatory cytokines.

  7. Human adipose tissue-derived mesenchymal stem cells expressing yeast cytosinedeaminase::uracil phosphoribosyltransferase inhibit intracerebral rat glioblastoma

    Czech Academy of Sciences Publication Activity Database

    Altanerova, V.; Cihova, M.; Babič, Michal; Rychly, B.; Ondicova, K.; Mravec, B.; Altaner, C.

    2012-01-01

    Roč. 130, č. 10 (2012), s. 2455-2463. ISSN 0020-7136 Institutional research plan: CEZ:AV0Z40500505 Keywords : glioblastoma * mesenchymal stem cells * suicide gene therapy Subject RIV: CD - Macromolecular Chemistry Impact factor: 6.198, year: 2012

  8. Construction of the Eukaryotic Expression Vector with EGFP and hVE GF121 Gene and its Expression in Rat Mesenchymal Stem Cells

    Institute of Scientific and Technical Information of China (English)

    Su Li; Chen Yunzhen; Zhang Xiaogang; She Qiang

    2005-01-01

    Objectives To construct a recombinant plasmid carrying enhanced green fluorescent protein (EGFP) and human vascular endothelial growth factor (VEGF) 121 gene and detect its expression in rat mesenchymal stem cells (MSCs). Methods Human VEGF121 cDNA was amplified with polymerase chain reaction (PCR) from pCD/hVEGF121 and was inserted into the eukaryotic expression vector pEGFPC1. After being identified with PCR, double enzyme digestion and DNA sequencing. The recombinant plasmid pEGFP/hVEGF121 was transferred into rat MSCs with lipofectamine. The expression of EGFP/VEGF121 fusion protein were detected with fluorescence microscope and immunocytochemical staining respectively. Results The recombinant plasmid was confirmed with PCR, double enzyme digestion and DNA sequencing. The fluorescence microscope and immunocytochemical staining results showed that the EGFP and VEGF121 protein were expressed in MSCs 48 h after transfection.Conclusions The recombinant plasmid carrying EGFP and human VEGF was successfully constructed and expressed positively in rat MSCs. It offers a promise tool for further research on differentiation of MSCs and VEGF gene therapy for ischemial cardiovascular disease.

  9. In Vitro Differentiation of Human Umbilical Cord Blood CD133+Cells into Insulin Producing Cells in Co-Culture with Rat Pancreatic Mesenchymal Stem Cells

    Science.gov (United States)

    Sahraneshin Samani, Fazel; Ebrahimi, Marzieh; Zandieh, Tahereh; Khoshchehreh, Reyhaneh; Baghaban Eslaminejad, Mohamadreza; Aghdami, Nasser; Baharvand, Hossein

    2015-01-01

    Objective Pancreatic stroma plays an important role in the induction of pancreatic cells by the use of close range signaling. In this respect, we presume that pancreatic mesenchymal cells (PMCs) as a fundamental factor of the stromal niche may have an effective role in differentiation of umbilical cord blood cluster of differentiation 133+ (UCB-CD133+) cells into newly-formed β-cells in vitro. Materials and Methods This study is an experimental research. The UCB-CD133+cells were purified by magnetic activated cell sorting (MACS) and differentiated into insulin producing cells (IPCs) in co-culture, both directly and indirectly with rat PMCs. Immunocytochemistry and enzyme linked immune sorbent assay (ELISA) were used to determine expression and production of insulin and C-peptide at the protein level. Results Our results demonstrated that UCB-CD133+differentiated into IPCs. Cells in islet-like clusters with (out) co-cultured with rat pancreatic stromal cells produced insulin and C-peptide and released them into the culture medium at the end of the induction protocol. However they did not respond well to glucose challenges. Conclusion Rat PMCs possibly affect differentiation of UCB-CD133+cells into IPCs by increasing the number of immature β-cells. PMID:26199900

  10. Human Umbilical Cord Blood-Derived Mesenchymal Stem Cell Therapy Promotes Functional Recovery of Contused Rat Spinal Cord through Enhancement of Endogenous Cell Proliferation and Oligogenesis

    Directory of Open Access Journals (Sweden)

    Sang In Park

    2012-01-01

    Full Text Available Numerous studies have shown the benefits of mesenchymal stem cells (MSCs on the repair of spinal cord injury (SCI model and on behavioral improvement, but the underlying mechanisms remain unclear. In this study, to investigate possible mechanisms by which MSCs contribute to the alleviation of neurologic deficits, we examined the potential effect of human umbilical cord blood-derived MSCs (hUCB-MSCs on the endogenous cell proliferation and oligogenesis after SCI. SCI was injured by contusion using a weight-drop impactor and hUCB-MSCs were transplanted into the boundary zone of the injured site. Animals received a daily injection of bromodeoxyuridine (BrdU for 7 days after treatment to identity newly synthesized cells of ependymal and periependymal cells that immunohistochemically resembled stem/progenitor cells was evident. Behavior analysis revealed that locomotor functions of hUCB-MSCs group were restored significantly and the cavity volume was smaller in the MSCs-transplanted rats compared to the control group. In MSCs-transplanted group, TUNEL-positive cells were decreased and BrdU-positive cells were significantly increased rats compared with control group. In addition, more of BrdU-positive cells expressed neural stem/progenitor cell nestin and oligo-lineage cell such as NG2, CNPase, MBP and glial fibrillary acidic protein typical of astrocytes in the MSC-transplanted rats. Thus, endogenous cell proliferation and oligogenesis contribute to MSC-promoted functional recovery following SCI.

  11. In Vitro Differentiation of Human Umbilical Cord Blood CD133+ Cells into Insulin Producing Cells in Co-Culture with Rat Pancreatic Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Fazel Sahraneshin Samani

    2015-07-01

    Full Text Available Objective: Pancreatic stroma plays an important role in the induction of pancreatic cells by the use of close range signaling. In this respect, we presume that pancreatic mesenchymal cells (PMCs as a fundamental factor of the stromal niche may have an effective role in differentiation of umbilical cord blood cluster of differentiation 133+ (UCB-CD133+ cells into newly-formed β-cells in vitro. Materials and Methods: This study is an experimental research. The UCB-CD133+ cells were purified by magnetic activated cell sorting (MACS and differentiated into insulin producing cells (IPCs in co-culture, both directly and indirectly with rat PMCs. Immunocytochemistry and enzyme linked immune sorbent assay (ELISA were used to determine expression and production of insulin and C-peptide at the protein level. Results: Our results demonstrated that UCB-CD133+ differentiated into IPCs. Cells in islet-like clusters with (out co-cultured with rat pancreatic stromal cells produced insulin and C-peptide and released them into the culture medium at the end of the induction protocol. However they did not respond well to glucose challenges. Conclusion: Rat PMCs possibly affect differentiation of UCB-CD133+ cells into IPCs by increasing the number of immature β-cells.

  12. Bone marrow mesenchymal stem cell transplantation combined with perindopril treatment attenuates infarction remodelling in a rat model of acute myocardial infarction

    Institute of Scientific and Technical Information of China (English)

    WANG Jian-an; LUO Rong-hua; ZHANG Xing; XIE Xiao-jie; HU Xin-yang; HE Ai-na; CHEN Jie; LI Jia-hui

    2006-01-01

    Objective: This study was performed to evaluate whether implantation of mesenchymal stem cell (MSC) would reduce left ventricular remodelling from the molecular mechanisms compared with angiotensin-converting enzyme inhibitors (ACEIs) perindopril into ischemic myocardium after acute myocardial infarction. Methods: Forty rats were divided into four groups: control, MSC, ACEI, MSC+ACEI groups. Bone marrow stem cell derived rat was injected immediately into a zone made ischemic by coronary artery ligation in MSC group and MSC+ACEI group. Phosphate-buffered saline (PBS) was injected into control group. Perindopril was administered p.o. to ACEI group and MSC+ACEI group. Six weeks after implantation, the rats were killed and heart sample was collected. Fibrillar collagen was observed by meliorative Masson's trichome stain. Western Blotting was employed to evaluate the protein expression of matrix metalloproteinase (MMP)-2, matrix metalloproteinase (MMP)-9 in infarction zone. The transcriptional level of MMP2, MMP9 and tissue inhibitor of matrix metalloproteinase (TIMP)-1in infarction area was detected by reverse transcriptase PCR (RT-PCR) analysis. Results: The fibrillar collagen area, the protein expression of MMP2, MMP9 and the transcriptional level of MMP2, MMP9 mRNA in infarction zone reduced in MSC group,ACEI group, and MSC+ACEI group. No significant difference was detected in the expression of TIMP1 mRNA among the 4groups. Conclusion: Both MSC and ACEI could reduce infarction remodelling by altering collagen metabolism.

  13. Intrathecally Transplanting Mesenchymal Stem Cells (MSCs) Activates ERK1/2 in Spinal Cords of Ischemia-Reperfusion Injury Rats and Improves Nerve Function.

    Science.gov (United States)

    Wang, Yonghong; Liu, He; Ma, Hong

    2016-01-01

    BACKGROUND We investigated whether an intrathecal transplantation of mesenchymal stem cells (MSCs) activates extracellular adjusting protein kinase1 and 2(ERK1/2) in the spinal cords of rats following an ischemia-reperfusion injury, resulting in improved spinal cord function and inhibition of apoptosis. MATERIAL AND METHODS We observed the relationship between the activation of ERK1/2 in the rat spinal cord and intrathecal transplantation of MSCs, as well as the effect of U0126, a MEK1/2 (upstream protein of ERK1/2) inhibitor, on a spinal cord ischemia-reperfusion injury model in rats using Basso Beattie Bresnahan (BBB) scoring, somatosensory evoked potentials (SSEPs), immunohistochemistry, and Western blot analysis. RESULTS After transplantation of MSCs, the lower limb motor function score increased, and the incubation period of SSEPs and amplitude were improved. Moreover, following transplantation of MSCs, Bcl2 expression increased, whereas Bax expression decreased after reperfusion. Transplantation of MSCs significantly enhanced pERK1/2 expression in the spinal cord, as well as pERK1/2 in immunoreactive cells located in the grey matter of the L4/5 levels of the spinal cord, following ischemia reperfusion injury in rats. The effective dose of U0126 required to inhibit pERK1/2 expression was 200 µg/kg. Bcl-2 decreased and the level of Bax expression increased in the spinal cord after ischemia reperfusion injury, and the protective effects of MSCs were attenuated. CONCLUSIONS Our findings suggest that intrathecal transplantation of MSCs activates ERK1/2 in the spinal cord following ischemia reperfusion injury, partially improves spinal cord function, and inhibits apoptosis in rats. PMID:27135658

  14. Intravenous administration of mesenchymal stem cells exerts therapeutic effects on parkinsonian model of rats: Focusing on neuroprotective effects of stromal cell-derived factor-1α

    Directory of Open Access Journals (Sweden)

    Tayra Judith

    2010-04-01

    Full Text Available Abstract Background Mesenchymal stem cells (MSCs are pluripotent stem cells derived from bone marrow with secretory functions of various neurotrophic factors. Stromal cell-derived factor-1α (SDF-1α is also reported as one of chemokines released from MSCs. In this research, the therapeutic effects of MSCs through SDF-1α were explored. 6-hydroxydopamine (6-OHDA, 20 μg was injected into the right striatum of female SD rats with subsequent administration of GFP-labeled MSCs, fibroblasts, (i.v., 1 × 107 cells, respectively or PBS at 2 hours after 6-OHDA injection. All rats were evaluated behaviorally with cylinder test and amphetamine-induced rotation test for 1 month with consequent euthanasia for immunohistochemical evaluations. Additionally, to explore the underlying mechanisms, neuroprotective effects of SDF-1α were explored using 6-OHDA-exposed PC12 cells by using dopamine (DA assay and TdT-mediated dUTP-biotin nick-end labeling (TUNEL staining. Results Rats receiving MSC transplantation significantly ameliorated behaviorally both in cylinder test and amphetamine-induced rotation test compared with the control groups. Correspondingly, rats with MSCs displayed significant preservation in the density of tyrosine hydroxylase (TH-positive fibers in the striatum and the number of TH-positive neurons in the substantia nigra pars compacta (SNc compared to that of control rats. In the in vitro study, SDF-1α treatment increased DA release and suppressed cell death induced by 6-OHDA administration compared with the control groups. Conclusions Consequently, MSC transplantation might exert neuroprotection on 6-OHDA-exposed dopaminergic neurons at least partly through anti-apoptotic effects of SDF-1α. The results demonstrate the potentials of intravenous MSC administration for clinical applications, although further explorations are required.

  15. Ultrasound-targeted stromal cell-derived factor-1-loaded microbubble destruction promotes mesenchymal stem cell homing to kidneys in diabetic nephropathy rats

    Directory of Open Access Journals (Sweden)

    Wu S

    2014-12-01

    Full Text Available Shengzheng Wu,1 Lu Li,1 Gong Wang,1 Weiwei Shen,2 Yali Xu,1 Zheng Liu,1 Zhongxiong Zhuo,1 Hongmei Xia,1 Yunhua Gao,1 Kaibin Tan1 1Department of Ultrasound, 2Department of Orthopedics, Xinqiao Hospital, Third Military Medical University, Chongqing, People’s Republic of China Abstract: Mesenchymal stem cell (MSC therapy has been considered a promising strategy to cure diabetic nephropathy (DN. However, insufficient MSCs can settle in injured kidneys, which constitute one of the major barriers to the effective implementation of MSC therapy. Stromal cell-derived factor-1 (SDF-1 plays a vital role in MSC migration and involves activation, mobilization, homing, and retention, which are presumably related to the poor homing in DN therapy. Ultrasound-targeted microbubble destruction has become one of the most promising strategies for the targeted delivery of drugs and genes. To improve MSC homing to DN kidneys, we present a strategy to increase SDF-1 via ultrasound-targeted microbubble destruction. In this study, we developed SDF-1-loaded microbubbles (MBSDF-1 via covalent conjugation. The characterization and bioactivity of MBSDF-1 were assessed in vitro. Target release in the targeted kidneys was triggered with diagnostic ultrasound in combination with MBSDF-1. The related bioeffects were also elucidated. Early DN was induced in rats with streptozotocin. Green fluorescent protein-labeled MSCs were transplanted intravenously following the target release of SDF-1 in the kidneys of normal and DN rats. The homing efficacy was assessed by detecting the implanted exogenous MSCs at 24 hours. The in vitro results showed an impressive SDF-1 loading efficacy of 79% and a loading content of 15.8 µg/mL. MBSDF-1 remained bioactive as a chemoattractant. In the in vivo study, SDF-1 was successfully released in the targeted kidneys. The homing efficacy of MSCs to DN kidneys after the target release of SDF-1 was remarkably ameliorated at 24 hours compared with

  16. Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide

    OpenAIRE

    Naaldijk Yahaira; Staude Marek; Fedorova Viktoriya; Stolzing Alexandra

    2012-01-01

    Abstract Background Mesenchymal stem cells (MSCs) are increasingly used as therapeutic agents as well as research tools in regenerative medicine. Development of technologies which allow storing and banking of MSC with minimal loss of cell viability, differentiation capacity, and function is required for clinical and research applications. Cryopreservation is the most effective way to preserve cells long term, but it involves potentially cytotoxic compounds and processing steps. Here, we inves...

  17. MicroRNA-9 promotes the neuronal differentiation of rat bone marrow mesenchymal stem cells by activating autophagy

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    Guang-yu Zhang

    2015-01-01

    Full Text Available MicroRNA-9 (miR-9 has been shown to promote the differentiation of bone marrow mesenchymal stem cells into neuronal cells, but the precise mechanism is unclear. Our previous study confirmed that increased autophagic activity improved the efficiency of neuronal differentiation in bone marrow mesenchymal stem cells. Accumulating evidence reveals that miRNAs adjust the autophagic pathways. This study used miR-9-1 lentiviral vector and miR-9-1 inhibitor to modulate the expression level of miR-9. Autophagic activity and neuronal differentiation were measured by the number of light chain-3 (LC3-positive dots, the ratio of LC3-II/LC3, and the expression levels of the neuronal markers enolase and microtubule-associated protein 2. Results showed that LC3-positive dots, the ratio of LC3-II/LC3, and expression of neuron specific enolase and microtubule-associated protein 2 increased in the miR-9 + group. The above results suggest that autophagic activity increased and bone marrow mesenchymal stem cells were prone to differentiate into neuronal cells when miR-9 was overexpressed, demonstrating that miR-9 can promote neuronal differentiation by increasing autophagic activity.

  18. MicroRNA-9 promotes the neuronal differentiation of rat bone marrow mesenchymal stem cells by activating autophagy

    Institute of Scientific and Technical Information of China (English)

    Guang-yu Zhang; Jun Wang; Yan-jie Jia; Rui Han; Ping Li; Deng-na Zhu

    2015-01-01

    MicroRNA-9 (miR-9) has been shown to promote the differentiation of bone marrow mesen-chymal stem cells into neuronal cells, but the precise mechanism is unclear. Our previous study conifrmed that increased autophagic activity improved the efifciency of neuronal differentiation in bone marrow mesenchymal stem cells. Accumulating evidence reveals that miRNAs adjust the autophagic pathways. This study used miR-9-1 lentiviral vector and miR-9-1 inhibitor to modulate the expression level of miR-9. Autophagic activity and neuronal differentiation were measured by the number of light chain-3 (LC3)-positive dots, the ratio of LC3-II/LC3, and the expression levels of the neuronal markers enolase and microtubule-associated protein 2. Re-sults showed that LC3-positive dots, the ratio of LC3-II/LC3, and expression of neuron speciifc enolase and microtubule-associated protein 2 increased in the miR-9+ group. The above results suggest that autophagic activity increased and bone marrow mesenchymal stem cells were prone to differentiate into neuronal cells when miR-9 was overexpressed, demonstrating that miR-9 can promote neuronal differentiation by increasing autophagic activity.

  19. Protective effect of adipose-derived mesenchymal stem cells against acute kidney injury induced by ischemia-reperfusion in Sprague-Dawley rats

    Science.gov (United States)

    SHEASHAA, HUSSEIN; LOTFY, AHMED; ELHUSSEINI, FATMA; AZIZ, AZZA ABDEL; BAIOMY, AZZA; AWAD, SAMAH; ALSAYED, AZIZA; EL-GILANY, ABDEL-HADY; SAAD, MOHAMED-AHDY A.A.; MAHMOUD, KHALED; ZAHRAN, FATEN; SALEM, DALIA A.; SARHAN, AHMED; GHAFFAR, HASSAN ABDEL; SOBH, MOHAMED

    2016-01-01

    Acute kidney injury (AKI) is a complex clinical condition associated with significant morbidity and mortality and lacking effective management. Ischemia-reperfusion injury (IRI) remains one of the leading causes of AKI in native and transplanted kidneys. The aim of this study was to evaluate the efficacy of adipose-derived mesenchymal stem cells (ADSCs) in the prevention of renal IRI in rats. The study was conducted on male Sprague-Dawley rats (n=72) weighing 250–300 g. Rats were randomly assigned to three main groups: i) Sham-operated control group (n=24); ii) positive control group, in which rats were subjected to IRI and were administered culture media following 4 h of IRI (n=24); and iii) ADSC group (n=24), in which rats were administered 1×106 ADSCs via the tail vein following 4 h of IRI. Each main group was further divided according to the timing after IRI into four equal-sized subgroups. Renal function was tested via the measurement of serum creatinine levels and creatinine clearance. In addition, malondialdehyde (MDA) levels were determined in serum and renal tissue homogenate as an indicator of oxidative stress. Histopathological changes were analyzed in different regions of the kidney, namely the cortex, outer stripe of the outer medulla (OSOM), inner stripe of the outer medulla (ISOM) and inner medulla. In each region, the scoring system considered active injury changes, regenerative changes and chronic changes. The ADSCs were assessed and their differentiation capability was verified. IRI resulted in a significant increase in serum creatinine, serum and tissue MDA levels and a significant reduction in creatinine clearance compared with those in sham-operated rats,. These changes were attenuated by the use of ADSCs. The prominent histopathological changes in the cortex, ISOM and OSOM were reflected in the injury score, which was significantly evident in the positive control group. The use of ADSCs was associated with significantly lowered injury

  20. Exosomes Secreted by Human-Induced Pluripotent Stem Cell-Derived Mesenchymal Stem Cells Repair Critical-Sized Bone Defects through Enhanced Angiogenesis and Osteogenesis in Osteoporotic Rats

    Science.gov (United States)

    Qi, Xin; Zhang, Jieyuan; Yuan, Hong; Xu, Zhengliang; Li, Qing; Niu, Xin; Hu, Bin; Wang, Yang; Li, Xiaolin

    2016-01-01

    Bone defects caused by trauma, severe infection, tumor resection and skeletal abnormalities are common osteoporotic conditions and major challenges in orthopedic surgery, and there is still no effective solution to this problem. Consequently, new treatments are needed to develop regeneration procedures without side effects. Exosomes secreted by mesenchymal stem cells (MSCs) derived from human induced pluripotent stem cells (hiPSCs, hiPSC-MSC-Exos) incorporate the advantages of both MSCs and iPSCs with no immunogenicity. However, there are no reports on the application of hiPSC-MSC-Exos to enhance angiogenesis and osteogenesis under osteoporotic conditions. HiPSC-MSC-Exos were isolated and identified before use. The effect of hiPSC-MSC-Exos on the proliferation and osteogenic differentiation of bone marrow MSCs derived from ovariectomized (OVX) rats (rBMSCs-OVX) in vitro were investigated. In vivo, hiPSC-MSC-Exos were implanted into critical size bone defects in ovariectomized rats, and bone regeneration and angiogenesis were examined by microcomputed tomography (micro-CT), sequential fluorescent labeling analysis, microfil perfusion and histological and immunohistochemical analysis. The results in vitro showed that hiPSC-MSC-Exos enhanced cell proliferation and alkaline phosphatase (ALP) activity, and up-regulated mRNA and protein expression of osteoblast-related genes in rBMSCs-OVX. In vivo experiments revealed that hiPSC-MSC-Exos dramatically stimulated bone regeneration and angiogenesis in critical-sized calvarial defects in ovariectomized rats. The effect of hiPSC-MSC-Exos increased with increasing concentration. In this study, we showed that hiPSC-MSC-Exos effectively stimulate the proliferation and osteogenic differentiation of rBMSCs-OVX, with the effect increasing with increasing exosome concentration. Further analysis demonstrated that the application of hiPSC-MSC-Exos+β-TCP scaffolds promoted bone regeneration in critical-sized calvarial defects by

  1. Application potential of bone marrow mesenchymal stem cell (BMSCs) based tissue-engineering for spinal cord defect repair in rat fetuses with spina bifida aperta.

    Science.gov (United States)

    Li, Xiaoshuai; Yuan, Zhengwei; Wei, Xiaowei; Li, Hui; Zhao, Guifeng; Miao, Jiaoning; Wu, Di; Liu, Bo; Cao, Songying; An, Dong; Ma, Wei; Zhang, Henan; Wang, Weilin; Wang, Qiushi; Gu, Hui

    2016-04-01

    Spina bifida aperta are complex congenital malformations resulting from failure of fusion in the spinal neural tube during embryogenesis. Despite surgical repair of the defect, most patients who survive with spina bifida aperta have a multiple system handicap due to neuron deficiency of the defective spinal cord. Tissue engineering has emerged as a novel treatment for replacement of lost tissue. This study evaluated the prenatal surgical approach of transplanting a chitosan-gelatin scaffold seeded with bone marrow mesenchymal stem cells (BMSCs) in the healing the defective spinal cord of rat fetuses with retinoic acid induced spina bifida aperta. Scaffold characterisation revealed the porous structure, organic and amorphous content. This biomaterial promoted the adhesion, spreading and in vitro viability of the BMSCs. After transplantation of the scaffold combined with BMSCs, the defective region of spinal cord in rat fetuses with spina bifida aperta at E20 decreased obviously under stereomicroscopy, and the skin defect almost closed in many fetuses. The transplanted BMSCs in chitosan-gelatin scaffold survived, grew and expressed markers of neural stem cells and neurons in the defective spinal cord. In addition, the biomaterial presented high biocompatibility and slow biodegradation in vivo. In conclusion, prenatal transplantation of the scaffold combined with BMSCs could treat spinal cord defect in fetuses with spina bifida aperta by the regeneration of neurons and repairmen of defective region. PMID:26894267

  2. New therapy of skin repair combining adipose-derived mesenchymal stem cells with sodium carboxymethylcellulose scaffold in a pre-clinical rat model.

    Directory of Open Access Journals (Sweden)

    Cristiano Rodrigues

    Full Text Available Lesions with great loss of skin and extensive burns are usually treated with heterologous skin grafts, which may lead rejection. Cell therapy with mesenchymal stem cells is arising as a new proposal to accelerate the healing process. We tested a new therapy consisting of sodium carboxymethylcellulose (CMC as a biomaterial, in combination with adipose-derived stem cells (ADSCs, to treat skin lesions in an in vivo rat model. This biomaterial did not affect membrane viability and induced a small and transient genotoxicity, only at the highest concentration tested (40 mg/mL. In a rat wound model, CMC at 10 mg/mL associated with ADSCs increased the rate of cell proliferation of the granulation tissue and epithelium thickness when compared to untreated lesions (Sham, but did not increase collagen fibers nor alter the overall speed of wound closure. Taken together, the results show that the CMC is capable to allow the growth of ADSCs and is safe for this biological application up to the concentration of 20 mg/mL. These findings suggest that CMC is a promising biomaterial to be used in cell therapy.

  3. Articular chondrocytes and mesenchymal stem cells seeded on biodegradable scaffolds for the repair of cartilage in a rat osteochondral defect model.

    Science.gov (United States)

    Dahlin, Rebecca L; Kinard, Lucas A; Lam, Johnny; Needham, Clark J; Lu, Steven; Kasper, F Kurtis; Mikos, Antonios G

    2014-08-01

    This work investigated the ability of co-cultures of articular chondrocytes and mesenchymal stem cells (MSCs) to repair articular cartilage in osteochondral defects. Bovine articular chondrocytes and rat MSCs were seeded in isolation or in co-culture onto electrospun poly(ɛ-caprolactone) (PCL) scaffolds and implanted into an osteochondral defect in the trochlear groove of 12-week old Lewis rats. Additionally, a blank PCL scaffold and untreated defect were investigated. After 12 weeks, the extent of cartilage repair was analyzed through histological analysis, and the extent of bone healing was assessed by quantifying the total volume of mineralized bone in the defect through microcomputed tomography. Histological analysis revealed that the articular chondrocytes and co-cultures led to repair tissue that consisted of more hyaline-like cartilage tissue that was thicker and possessed more intense Safranin O staining. The MSC, blank PCL scaffold, and empty treatment groups generally led to the formation of fibrocartilage repair tissue. Microcomputed tomography revealed that while there was an equivalent amount of mineralized bone formation in the MSC, blank PCL, and empty treatment groups, the defects treated with chondrocytes or co-cultures had negligible mineralized bone formation. Overall, even with a reduced number of chondrocytes, co-cultures led to an equal level of cartilage repair compared to the chondrocyte samples, thus demonstrating the potential for the use of co-cultures of articular chondrocytes and MSCs for the in vivo repair of cartilage defects. PMID:24927682

  4. In vivo tracing of superparamagnetic iron oxide-labeled bone marrow mesenchymal stem cells transplanted for traumatic brain injury by susceptibility weighted imaging in a rat model

    Institute of Scientific and Technical Information of China (English)

    CHENG Jing-liang; YANG Yun-jun; LI Hua-li; WANG Juan; WANG Mei-hao; ZHANG Yong

    2010-01-01

    Objective:To label rat bone marrow mesenchymal stem cells (BMSCs) with superparamagnetic iron oxide (SPIO) in vitro, and to monitor the survival and location of these labeled BMSCs in a rat model of traumatic brain injury (TBI) by susceptibility weighted imaging (SWI)sequence.Methods:BMSCs were cultured in vitro and then labeled with SPIO. Totally 24 male Sprague Dawley (SD) rats weighing 200-250 g were randomly divided into 4 groups: Groups A-D (n=6 for each group). Moderate TBI models of all the rats were developed in the left hemisphere following Feeney's method. Group A was the experimental group and stereotaxic transplantation of BMSCs labeled with SPIO into the region nearby the contusion was conducted in this group 24 hours after TBI modeling. The other three groups were control groups with transplantation of SPIO, unlabeled BMSCs and injection of nutrient solution respectively conducted in Groups B, C and D at the same time. Monitoring of these SPIO-labeled BMSCs by SWI was performed one day,one week and three weeks after implantation.Results: Numerous BMSCs were successfully labeled with SPIO. They were positive for Prussian blue staining and intracytoplasm positive blue stained particles were found under a microscope (×200). Scattered little iron particles were observed in the vesicles by electron microscopy (×5000). MRI of the transplantation sites of the left hemisphere demonstrated a low signal intensity on magnitude images,phase images and SWI images for all the test rats in Group A, and the lesion in the left parietal cortex demonstrated a semicircular low intensity on SWI images, which clearly showed the distribution and migration of BMSCs in the first and third weeks. For Group B, a low signal intensity by MRI was only observed on the first day but undetected during the following examination. No signals were observed in Groups C and D at any time points.Conclusion:SWI sequence in vivo can consecutively and noninvasively trace and demonstrate the

  5. Hypoxia/Reoxygenation-Preconditioned Human Bone Marrow-Derived Mesenchymal Stromal Cells Rescue Ischemic Rat Cortical Neurons by Enhancing Trophic Factor Release.

    Science.gov (United States)

    Kim, Young Seo; Noh, Min Young; Cho, Kyung Ah; Kim, Hyemi; Kwon, Min-Soo; Kim, Kyung Suk; Kim, Juhan; Koh, Seong-Ho; Kim, Seung Hyun

    2015-08-01

    Bone marrow-derived mesenchymal stromal cells (BM-MSCs) represent a promising tool for stem cell-based therapies. However, the majority of MSCs fail to reach the injury site and have only minimal therapeutic effect. In this study, we assessed whether hypoxia/reoxygenation (H/R) preconditioning of human BM-MSCs could increase their functional capacity and beneficial effect on ischemic rat cortical neurons. Human BM-MSCs were cultured under hypoxia (1% O2) and with long-term reoxygenation for various times to identify the optimal conditions for increasing their viability and proliferation. The effects of H/R preconditioning on the BM-MSCs were assessed by analyzing the expression of prosurvival genes, trophic factors, and cell migration assays. The functionally improved BM-MSCs were cocultured with ischemic rat cortical neurons to compare with normoxic cultured BM-MSCs. Although the cell viability and proliferation of BM-MSCs were reduced after 1 day of hypoxic culture (1% O2), when this was followed by 5-day reoxygenation, the BM-MSCs recovered and multiplied extensively. The immunophenotype and trilineage differentiation of BM-MSCs were also maintained under this H/R preconditioning. In addition, the preconditioning enhanced the expression of prosurvival genes, the messenger RNA (mRNA) levels of various trophic factors and migration capacity. Finally, coculture with the H/R-preconditioned BM-MSCs promoted the survival of ischemic rat cortical neurons. H/R preconditioning of BM-MSCs increases prosurvival signals, trophic factor release, and cell migration and appears to increase their ability to rescue ischemic cortical neurons. This optimized H/R preconditioning procedure could provide the basis for a new strategy for stem cell therapy in ischemic stroke patients. PMID:25288154

  6. Intravenous transplantation of bone marrow mesenchymal stem cells promotes neural regeneration after traumatic brain injury

    OpenAIRE

    Anbari, Fatemeh; Khalili, Mohammad Ali; Bahrami, Ahmad Reza; Khoradmehr, Arezoo; Sadeghian, Fatemeh; Fesahat, Farzaneh; Nabi, Ali

    2014-01-01

    To investigate the supplement of lost nerve cells in rats with traumatic brain injury by intravenous administration of allogenic bone marrow mesenchymal stem cells, this study established a Wistar rat model of traumatic brain injury by weight drop impact acceleration method and administered 3 × 106 rat bone marrow mesenchymal stem cells via the lateral tail vein. At 14 days after cell transplantation, bone marrow mesenchymal stem cells differentiated into neurons and astrocytes in injured rat...

  7. Biofunctionalization of a titanium surface with a nano-sawtooth structure regulates the behavior of rat bone marrow mesenchymal stem cells

    Directory of Open Access Journals (Sweden)

    Zhang WJ

    2012-08-01

    Full Text Available Wenjie Zhang,1,2 Zihui Li,3 Yan Liu,1,2 Dongxia Ye,4 Jinhua Li,3 Lianyi Xu,1,2 Bin Wei,1 Xiuli Zhang,2 Xuanyong Liu,3,* Xinquan Jiang,1,2,* 1Department of Prosthodontics, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, 2Oral Bioengineering Laboratory, Shanghai Research Institute of Stomatology, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai Key Laboratory of Stomatology, 3State Key Laboratory of High Performance Ceramics and Superfine Microstructure, Shanghai Institute of Ceramics, Chinese Academy of Sciences, 4Shanghai Research Institute of Stomatology, Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University, School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai, China*Joint principal authors of this workBackground: The topography of an implant surface can serve as a powerful signaling cue for attached cells and can enhance the quality of osseointegration. A series of improved implant surfaces functionalized with nanoscale structures have been fabricated using various methods. Methods: In this study, using an H2O2 process, we fabricated two size-controllable sawtooth-like nanostructures with different dimensions on a titanium surface. The effects of the two nano-sawtooth structures on rat bone marrow mesenchymal stem cells (BMMSCs were evaluated without the addition of osteoinductive chemical factors.Results: These new surface modifications did not adversely affect cell viability, and rat BMMSCs demonstrated a greater increase in proliferation ability on the surfaces of the nano-sawtooth structures than on a control plate. Furthermore, upregulated expression of osteogenic-related genes and proteins indicated that the nano-sawtooth structures promote osteoblastic differentiation of rat BMMSCs. Importantly, the large nano-sawtooth structure resulted in the greatest cell responses, including increased adhesion, proliferation

  8. Magnetic resonance imaging tracing of transplanted bone marrow mesenchymal stem cells in a rat model of cardiac arrest-induced global brain ischemia

    Institute of Scientific and Technical Information of China (English)

    Yue Fu; Xiangshao Fang; Tong Wang; Jiwen Wang; Jun Jiang; Zhigang Luo; Xiaohui Duan; Jun Shen; Zitong Huang

    2009-01-01

    BACKGROUND: Numerous studies have shown that magnetic resonance imaging (MRI) can detect survival and migration of super paramagnetic iron oxide-labeled stem cells in models of focal cerebral infarction. OBJECTIVE: To observe distribution of bone marrow mesenchymal stem cells (BMSCs) in a rat model of global brain ischemia following cardiac arrest and resuscitation, and to investigate the feasibility of tracing iron oxide-labeled BMSCs using non-invasive MRI. DESIGN, TIME AND SETTING: The randomized, controlled, molecular imaging study was performed at the Linbaixin Medical Research Center, Second Affiliated Hospital, Sun Yat-sen University, and the Institute of Cardiopulmonary Cerebral Resuscitation, Sun Yat-sen University, China from October 2006 to February 2009.MATERIALS: A total of 40 clean, Sprague Dawley rats, aged 6 weeks and of either gender, were supplied by the Experimental Animal Center, Sun Yat-sen University, China, for isolation of BMSCs. Feridex (iron oxide), Gyroscan Inetra 1.5T MRI system, and cardiopulmonary resuscitation device were used in this study. METHODS: A total of 30 healthy, male Sprague Dawley rats, aged 6 months, were used to induce ventricular fibrillation using alternating current. After 8 minutes, the rats underwent 6-minute chest compression and mechanical ventilation, followed by electric defibrillation, to establish rat models of global brain ischemia due to cardiac arrest and resuscitation. A total of 24 successful models were randomly assigned to Feridex-labeled and non-labeled groups (n=12 for each group). At 2 hours after resuscitation, 5 x 10 6 Feddex-labeled BMSCs, with protamine sulfate as a carrier, and 5 × 10 6 non-labeled BMSCs were respectively transplanted into both groups of rats through the right carotid artery (cells were harvested in 1 mL phosphate buffered saline). MAIN OUTCOME MEASURES: Feridex-labeled BMSCs were observed by Prussian blue staining and electron microscopy. Signal intensity, celluar viability

  9. Effect of mesenchymal stem cells transplantation combining with hyperbaric oxygen therapy on rehabilitation of rat spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    Cheng-Kui Geng; Hong-Hua Cao; Xiong Ying; Hua-Lin Yu

    2015-01-01

    Objective:To investigate the effect of BMSCs transplantation plus hyperbaric oxygen (HBO) on repair of rat SCI. Methods:Seventy five male rats were divided randomly into five groups:sham, vehicle, BMSCs transplantation group, combination group, 15 rats in each group. Every week after the SCI onset, all animals were evaluated for behavior outcome by Basso-Beattle-Bresnahan (BBB) score and inclined plane test. Axon recovery was examined with focal spinal cord tissue by electron microscope at 6 weeks after the SCI onset. HE staining and BrdU staining were performed to examine the BMSCs and lesion post injury. Somatosensory evoked potential (SEP) testing was performed to detect the recovery of neural conduction. Results:Results from the behavior tests from combination group were significant higher than rats which received only transplantation or HBO treatment. Results from histopathology showed favorable recovery from combination group than other treatment groups. The number of BrdU+ in combination group were measureable more than transplantation group (P<0.05). The greatest decrease in TNF-α, IL-1β, IL-6, IFN-αdetermined by Elisa assay in combination group were evident too. Conclusions:BMSCs transplantation can promote the functional recovery of rat hind limbs after SCI, and its combination with HBO has a synergistic effect.

  10. Effect of mesenchymal stem cells transplantation combining with hyperbaric oxygen therapy on rehabilitation of rat spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    Cheng-Kui; Geng; Hong-Hua; Cao; Xiong; Ying; Hua-Lin; Yu

    2015-01-01

    Objective:To investigate the effect of BMSCs transplantation plus hyperbaric oxygen(HBO)on repair of rat SCI.Methods:Seventy five male rats were divided randomly into five groups:sham,vehicle.BMSCs transplantation group,combination group,15 rats in each group.Every week after the SCI onset,all animals were evaluated for behavior outcome by Basso-BeattleBresnahan(BBB) score and inclined plane test.Axon recovery was examined with focal spinal cord tissue by electron microscope at 6 weeks after the SCI onset.HE staining and BrdU staining were performed to examine the BMSCs and lesion post injury.Somatosensory evoked potential(SEP) testing was performed to detect the recovery of neural conduction.Results from the behavior tests from combination group were significant higher than rats which received only transplantation or HBO treatment.Results from histopathology showed favorable recovery from combination group than other treatment groups.The number of BrdU+ in combination group were measureable more than transplantation group(P<0.05).The greatest decrease in TNF-α,IL-1β,IL-6.IFN-α determined by Elisa assay in combination group were evident too.Conclusions:BMSCs transplantation can promote the functional recovery of rat hind limbs after SCI,and its combination with HBO has a synergistic effect.

  11. The Chemopreventive Effect of Tamoxifen Combined with Celecoxib on DMBA chemically-Induced Breast Cancer

    Institute of Scientific and Technical Information of China (English)

    Xiaoxu Liu; Huafeng Kang; Xijing Wang; Zhijun Dai; Fengjie Xue; Xinghuan Xue

    2007-01-01

    Objective: To investigate the chemopreventive effect of tamoxifen combined with a COX-2 selective inhibitor, celecoxib, on breast cancer in rats chemically induced by 7,12-dimethylben (a)anthracene (DMBA). Methods:DMBA was irrigated into the stomaches of SD female rats to build breast cancer model. A total of 120 rats were divided into four groups: control group, tamoxifen group, celecoxib group and combined group. The incidence rate, latent period, number and volume of breast cancer were detected and analyzed. Results:The tumor incidence rate of tamoxifen group (48.15%, 13/27) and celecoxib group (50.00%,14/28) were lower than that of control group (85.71%, 24/28), but higher than that of combined group (21.43%, 6/28). The tumor's latent period of tamoxifen group (97.54±1.85 d) and celecoxib group (96.79±2.89 d) were longer than that of control group (89.50±5.99 d), but shorter than that of combined group (103.67±3.39 d). The average tumor number of tamoxifen group (1.77±0.73) and celecoxib group (1.71±0.61) were less than that of control group (3.50±1.62), but more than that of combined group ( 1.17±0.42 ). The average tumor volume of tamoxifen group (1.78±0.71 cm3) and celecoxib group (2.05±1.04 cm3) were smaller than that of control group (6.42±3.96 cm3), but bigger than that of combined group (0.71±0.96 cm3) (P < 0.05 respectively).Conclusion:Celecoxib and tamoxifen are effective drugs in preventing the occurrence of rat breast cancer chemically induced by DMBA. Furthermore, combination of them has better chemopreventive effect.

  12. Mesenchymal Stem Cell Attenuates Neutrophil-predominant Inflammation and Acute Lung Injury in an In Vivo Rat Model of Ventilator-induced Lung Injury

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    Tian-Shun Lai

    2015-01-01

    Full Text Available Background: Subsequent neutrophil (polymorphonuclear neutrophil [PMN]-predominant inflammatory response is a predominant feature of ventilator-induced lung injury (VILI, and mesenchymal stem cell (MSC can improve mice survival model of endotoxin-induced acute lung injury, reduce lung impairs, and enhance the repair of VILI. However, whether MSC could attenuate PMN-predominant inflammatory in the VILI is still unknown. This study aimed to test whether MSC intervention could attenuate the PMN-predominate inflammatory in the mechanical VILI. Methods: Sprague-Dawley rats were ventilated for 2 hours with large tidal volume (20 mL/kg. MSCs were given before or after ventilation. The inflammatory chemokines and gas exchange were observed and compared dynamically until 4 hours after ventilation, and pulmonary pathological change and activation of PMN were observed and compared 4 hours after ventilation. Results: Mechanical ventilation (MV caused significant lung injury reflected by increasing in PMN pulmonary sequestration, inflammatory chemokines (tumor necrosis factor-alpha, interleukin-6 and macrophage inflammatory protein 2 in the bronchoalveolar lavage fluid, and injury score of the lung tissue. These changes were accompanied with excessive PMN activation which reflected by increases in PMN elastase activity, production of radical oxygen series. MSC intervention especially pretreatment attenuated subsequent lung injury, systemic inflammation response and PMN pulmonary sequestration and excessive PMN activation initiated by injurious ventilation. Conclusions: MV causes profound lung injury and PMN-predominate inflammatory responses. The protection effect of MSC in the VILI rat model is related to the suppression of the PMN activation.

  13. Mesenchymal Stem Cell Attenuates Neutrophil-predominant Inflammation and Acute Lung Injury in an In Vivo Rat Model of Ventilator-induced Lung Injury

    Institute of Scientific and Technical Information of China (English)

    Tian-Shun Lai; Zhi-Hong Wang; Shao-Xi Cai

    2015-01-01

    Background:Subsequent neutrophil (polymorphonuclear neutrophil [PMN])-predominant inflammatory response is a predominant feature of ventilator-induced lung injury (VILI),and mesenchymal stem cell (MSC) can improve mice survival model of endotoxin-induced acute lung injury,reduce lung impairs,and enhance the repair of VILI.However,whether MSC could attenuate PMN-predominant inflammatory in the VILI is still unknown.This study aimed to test whether MSC intervention could attenuate the PMN-predominate inflammatory in the mechanical VILI.Methods:Sprague-Dawley rats were ventilated for 2 hours with large tidal volume (20 mL/kg).MSCs were given before or after ventilation.The inflammatory chemokines and gas exchange were observed and compared dynamically until 4 hours after ventilation,and pulmonary pathological change and activation of PMN were observed and compared 4 hours after ventilation.Results:Mechanical ventilation (MV) caused significant lung injury reflected by increasing in PMN pulmonary sequestration,inflammatory chemokines (tumor necrosis factor-alpha,interleukin-6 and macrophage inflammatory protein 2) in the bronchoalveolar lavage fluid,and injury score of the lung tissue.These changes were accompanied with excessive PMN activation which reflected by increases in PMN elastase activity,production of radical oxygen series.MSC intervention especially pretreatment attenuated subsequent lung injury,systemic inflammation response and PMN pulmonary sequestration and excessive PMN activation initiated by injurious ventilation.Conclusions:MV causes profound lung injury and PMN-predominate inflammatory responses.The protection effect of MSC in the VILI rat model is related to the suppression of the PMN activation.

  14. Inhibition of the osteogenic differentiation of mesenchymal stem cells derived from the offspring of rats treated with caffeine during pregnancy and lactation.

    Science.gov (United States)

    Reis, Amanda Maria Sena; Ocarino, Natália de Melo; Boeloni, Jankerle Neves; Gomes, Dawidson Assis; Goes, Alfredo Miranda; Ferreira, Andrea da Fonseca; Serakides, Rogéria

    2016-03-01

    Caffeine is an alkaloid that is widely consumed due to its presence in drugs, coffee, tea, and chocolate. This compound passes to offspring through the placenta and milk; can cause teratogenic mutations; and reduces the formation, growth, and mass of bone. Because mesenchymal stem cells (MSCs) are responsible for generating the entire skeleton, we hypothesized that these cells are targets of caffeine. This study evaluated the osteogenic differentiation of MSCs derived from the offspring of rats treated with caffeine during pregnancy and lactation. Twenty-four adult Wistar rats were randomly divided into four groups, including one control group and three experimental groups treated with 25, 50, or 100 mg/kg of caffeine. At weaning, three 21-day-old pups from each dam in each group were euthanized for extraction of bone marrow cells for in vitro tests. Caffeine doses of 50 and 100 mg/kg significantly reduced the activity of alkaline phosphatase at 7, 14, and 21 days and the expression of collagen I at 21 days. However, the expression of gene transcripts for alkaline phosphatase, Runx-2, and bone sialoprotein, as well as the synthesis of mineralization nodules, decreased significantly in all groups treated with caffeine. The expression of osteocalcin was significantly reduced only in the group treated with 50 mg/kg caffeine. The caffeine that passes from the mother to the offspring during pregnancy and lactation reduces the osteogenic differentiation of MSCs. We propose that this reduction in the osteogenic potential of MSCs may be involved in the pathogenesis of osteopenia resulting from caffeine consumption. PMID:26634797

  15. Preconditioning in lowered oxygen enhances the therapeutic potential of human umbilical mesenchymal stem cells in a rat model of spinal cord injury.

    Science.gov (United States)

    Zhilai, Zhou; Biling, Mo; Sujun, Qiu; Chao, Dong; Benchao, Shi; Shuai, Huang; Shun, Yao; Hui, Zhang

    2016-07-01

    Human umbilical cord mesenchymal stem cells (UCMSCs) have recently been shown to hold great therapeutic potential for the treatment of spinal cord injury (SCI). However, the number of engrafted cells has been shown to decrease dramatically post-transplantation. Physioxia is known to enhance the paracrine properties and immune modulation of stem cells, a notion that has been applied in many clinical settings. We therefore hypothesized that preconditioning of UCMSCs in physioxic environment would enhance the regenerative properties of these cells in the treatment of rat SCI. UCMSCs were pretreated with either atmospheric normoxia (21% O2, N-UCMSC) or physioxia (5% O2, P-UCMSC). The MSCs were characterized using flow cytometry, immunocytochemistry, and real-time polymerase chain reaction. Furthermore, 10(5) N-UCMSC or P-UCMSC were injected into the injured spinal cord immediately after SCI, and locomotor function as well as cellular, molecular and pathological changes were compared between the groups. We found that N-UCMSC and P-UCMSC displayed similar surface protein expression. P-UCMSC grew faster, while physioxia up-regulated the expression of trophic and growth factors, including hepatocyte growth factor (HGF), brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor(VEGF), in UCMSCs. Compared to N-UCMSC, treatment with P-UCMSC was associated with marked changes in the SCI environment, with a significant increase in axonal preservation and a decrease in the number of caspase-3+ cells and ED-1+ macrophages. These changes were accompanied by improved functional recovery. Thus, the present study indicated that preculturing UCMSCs under 5% lowered oxygen physioxic conditions prior to transplantation improves their therapeutic potential for the treatment of SCI in rats. PMID:27085204

  16. Conditioned medium from bone marrow-derived mesenchymal stem cells improves recovery after spinal cord injury in rats: an original strategy to avoid cell transplantation.

    Directory of Open Access Journals (Sweden)

    Dorothée Cantinieaux

    Full Text Available Spinal cord injury triggers irreversible loss of motor and sensory functions. Numerous strategies aiming at repairing the injured spinal cord have been studied. Among them, the use of bone marrow-derived mesenchymal stem cells (BMSCs is promising. Indeed, these cells possess interesting properties to modulate CNS environment and allow axon regeneration and functional recovery. Unfortunately, BMSC survival and differentiation within the host spinal cord remain poor, and these cells have been found to have various adverse effects when grafted in other pathological contexts. Moreover, paracrine-mediated actions have been proposed to explain the beneficial effects of BMSC transplantation after spinal cord injury. We thus decided to deliver BMSC-released factors to spinal cord injured rats and to study, in parallel, their properties in vitro. We show that, in vitro, BMSC-conditioned medium (BMSC-CM protects neurons from apoptosis, activates macrophages and is pro-angiogenic. In vivo, BMSC-CM administered after spinal cord contusion improves motor recovery. Histological analysis confirms the pro-angiogenic action of BMSC-CM, as well as a tissue protection effect. Finally, the characterization of BMSC-CM by cytokine array and ELISA identified trophic factors as well as cytokines likely involved in the beneficial observed effects. In conclusion, our results support the paracrine-mediated mode of action of BMSCs and raise the possibility to develop a cell-free therapeutic approach.

  17. High-Dose {sup 111}In Induces G1 Cell Cycle Arrest and Cell Death in Rat Bone Marrow Mesenchymal Stem Cells

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    Park, Bok Nam; Shim, Woo Young; Ahn, Young Hwan; Lee, Jae Ho; Yoon, Joon Kee [Ajou Univ. School of Medicine, Suwon (Korea, Republic of)

    2012-06-15

    This study was performed to evaluate the effect of {sup 111}In-labeling on the cell growth, cycle and viability of bone marrow mesenchymal stem cells (BMSCs). Rat BMSCs were labeled with various doses of {sup 111}In (0.4-11.1 Bq/cell). The growth curve of {sup 111}In-BMSCs was obtained up to 14th day of labeling. The cell cycle was evaluated by 5-bromo-2-deoxyuridine (BrdU) labeling or prospidium iodide (PI) staining. Senescent cells were counted under a light microscope after staining with 5-bromo-4-chloro-3-indolyl-{sup D-}galactopyranoside. Flow cytometry was performed to measure apoptotic and necrotic fractions after staining with annexin V-FITC and PI. The growth of BMSCs labeled with higher doses of {sup 111}In (4.4 or 11.1 Bq/cell) was significantly inhibited from the 3rd day of labeling. Flow cytometry revealed less BrdU-positive BMSCs at 11.1 Bq {sup 111}In/cell (9.07%/3.18%) on the 14th day (control=1.60%/0.39%). However, no cellular senescence was visualized up to the 14th day. A high dose of {sup 111}In-labeling induced cell cycle arrest and death in BMSCs; therefore, it should be used with a careful dosimetry in case of applying it to humans.

  18. The Effect of Vitamin E on the In Vitro Differentiation of Adult Rat Bone Marrow Mesenchymal Stem Cells to Osteoblast During Sodium Arsenite Exposure

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    M. Soleimani Mehranjani

    2016-01-01

    Full Text Available Introduction & Objective: Sodium arsenite disturbs the differentiation of adult rat bone marrow mesenchymal stem cells (rMSCs to Osteoblast through oxidative stress. We aimed to investigate the preventive effect of vitamin E, a strong antioxidant, in sodium arsenite toxicity on rMSCs differentiation to osteoblast. Materials & Methods: rMSCs were cultured in Dulbecco’s Modified Eagles Medium containing 15% Fetal Bovine Serum and divided into: control, sodium arsenite (20 nM, vitamin E (50 µM and sodium arsenite + vitamin E for 21 days in the osteogenic media containing 10% of fetal bovine serum. Cell viability, bone matrix mineralization, intercellular and extracellular calcium, alkaline phosphatase activity, DNA damage and cell morphological changes were evaluated. Data were analyzed using one-way ANOVA and Tukey's test and means were considered significantly different at P<0.05. Results: Cell viability, bone matrix mineralization, calcium deposition, alkaline phosphatase activity and nuclei diameter decreased significantly in the sodium arsenite group. The mentioned parameters increased significantly in cells treated with sodium arsenite + vitamin E to the control level (P<0.05. Cytoplasmic extensions were also observed in the vitamin E group. Conclusions: Vitamin E reduces sodium arsenite toxicity, increasing osteogenic differentiation in rMSCs. Sci J Hamadan Univ Med Sci . 2016; 22 (4 :276-285

  19. The use of SHP-2 gene transduced bone marrow mesenchymal stem cells to promote osteogenic differentiation and bone defect repair in rat.

    Science.gov (United States)

    Fan, Dapeng; Liu, Shen; Jiang, Shichao; Li, Zhiwei; Mo, Xiumei; Ruan, Hongjiang; Zou, Gang-Ming; Fan, Cunyi

    2016-08-01

    Bone tissue engineering is a promising approach for bone regeneration, in which growth factors play an important role. The tyrosine phosphatase Src-homology region 2-containing protein tyrosine phosphatase 2 (SHP2), encoded by the PTPN11 gene, is essential for the differentiation, proliferation and metabolism of osteoblasts. However, SHP-2 has never been systematically studied for its effect in osteogenesis. We predicted that overexpression of SHP-2 could promote bone marrow-derived mesenchymal stem cell (BMSC)osteogenic differentiation and SHP-2 transduced BMSCs could enhance new bone formation, determined using the following study groups: (1) BMSCs transduced with SHP-2 and induced with osteoblast-inducing liquid (BMSCs/SHP-2/OL); (2) BMSCs transduced with SHP-2 (BMSCs/-SHP-2); (3) BMSCs induced with osteoblast-inducing liquid (BMSCs/OL) and (4) pure BMSCs. Cells were assessed for osteogenic differentiation by quantitative real-time polymerase chain reaction analysis, western blot analysis, alkaline phosphatase activity and alizarin red S staining. For in vivo assessment, cells were combined with beta-tricalcium phosphate scaffolds and transplanted into rat calvarial defects for 8 weeks. Following euthanasia, skull samples were explanted for osteogenic evaluation, including micro-computed tomography measurement, histology and immunohistochemistry staining. SHP-2 and upregulation of its gene promoted BMSC osteogenic differentiation and therefore represents a potential new therapeutic approach to bone repair. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1871-1881, 2016. PMID:26999642

  20. Combined Effects of Mechanical Strain and Hydroxyapatite/Collagen Composite on Osteogenic Differentiation of Rat Bone Marrow Derived Mesenchymal Stem Cells

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    Yan Huang

    2013-01-01

    Full Text Available Mesenchymal stem cells (MSCs represent a promising source for bone repair and regeneration. Recent lines of evidence have shown that appropriate strain could regulate the osteogenic differentiation of MSCs. Our previous study demonstrated that hydroxyapatite/collagen (HA/Col composite also played an important role in the osteogenic differentiation of MSCs. The aim of this study is to investigate the effects of mechanical strain and HA/Col composite on the osteogenic differentiation of rat bone marrow derived MSCs (rBMSCs in vitro. rBMSCs were treated with cyclic strain generated by a self-designed stretching device with or without the presence of HA/Col composite. Osteogenic differentiation levels were evaluated using reverse transcription polymerase chain reaction (RT-PCR, alkaline phosphatase spectrophotometry, and western blotting. The results demonstrated that mechanical strain combined with HA/Col composite could obviously induce the differentiation of rBMSCs into osteoblasts, which had a better effect than only mechanical strain or HA/Col composite treatment. This provides a new avenue for mechanistic studies of stem cell differentiation and a novel approach to obtain more committed differentiated cells.

  1. Nanoparticle Labeling of Bone Marrow-Derived Rat Mesenchymal Stem Cells: Their Use in Differentiation and Tracking

    Directory of Open Access Journals (Sweden)

    Ece Akhan

    2015-01-01

    Full Text Available Mesenchymal stem cells (MSCs are promising candidates for cellular therapies due to their ability to migrate to damaged tissue without inducing immune reaction. Many techniques have been developed to trace MSCs and their differentiation efficacy; however, all of these methods have limitations. Conjugated polymer based water-dispersible nanoparticles (CPN represent a new class of probes because they offer high brightness, improved photostability, high fluorescent quantum yield, and noncytotoxicity comparing to conventional dyes and quantum dots. We aimed to use this tool for tracing MSCs’ fate in vitro and in vivo. MSC marker expression, survival, and differentiation capacity were assessed upon CPN treatment. Our results showed that after CPN labeling, MSC markers did not change and significant number of cells were found to be viable as revealed by MTT. Fluorescent signals were retained for 3 weeks after they were differentiated into osteocytes, adipocytes, and chondrocytes in vitro. We also showed that the labeled MSCs migrated to the site of injury and retained their labels in an in vivo liver regeneration model. The utilization of nanoparticle could be a promising tool for the tracking of MSCs in vivo and in vitro and therefore can be a useful tool to understand differentiation and homing mechanisms of MSCs.

  2. In-vivo generation of bone via endochondral ossification by in-vitro chondrogenic priming of adult human and rat mesenchymal stem cells

    LENUS (Irish Health Repository)

    Farrell, Eric

    2011-01-31

    Abstract Background Bone grafts are required to repair large bone defects after tumour resection or large trauma. The availability of patients\\' own bone tissue that can be used for these procedures is limited. Thus far bone tissue engineering has not lead to an implant which could be used as alternative in bone replacement surgery. This is mainly due to problems of vascularisation of the implanted tissues leading to core necrosis and implant failure. Recently it was discovered that embryonic stem cells can form bone via the endochondral pathway, thereby turning in-vitro created cartilage into bone in-vivo. In this study we investigated the potential of human adult mesenchymal stem cells to form bone via the endochondral pathway. Methods MSCs were cultured for 28 days in chondrogenic, osteogenic or control medium prior to implantation. To further optimise this process we induced mineralisation in the chondrogenic constructs before implantation by changing to osteogenic medium during the last 7 days of culture. Results After 8 weeks of subcutaneous implantation in mice, bone and bone marrow formation was observed in 8 of 9 constructs cultured in chondrogenic medium. No bone was observed in any samples cultured in osteogenic medium. Switch to osteogenic medium for 7 days prevented formation of bone in-vivo. Addition of β-glycerophosphate to chondrogenic medium during the last 7 days in culture induced mineralisation of the matrix and still enabled formation of bone and marrow in both human and rat MSC cultures. To determine whether bone was formed by the host or by the implanted tissue we used an immunocompetent transgenic rat model. Thereby we found that osteoblasts in the bone were almost entirely of host origin but the osteocytes are of both host and donor origin. Conclusions The preliminary data presented in this manuscript demonstrates that chondrogenic priming of MSCs leads to bone formation in vivo using both human and rat cells. Furthermore, addition of

  3. Effect of mesenchymal stromal cells (MSC) on chronic visceral hypersensitivity in a radio-induced colonic ulceration model in the rat

    International Nuclear Information System (INIS)

    Patients who undergo pelvic radiotherapy may develop significant incidence of undesirable chronic gastrointestinal complications resulting from radiation-induced damages around the tumour. Chronic visceral pain is one of the radiation-induced side effects that greatly affects the quality of life of 'cancer survivors'. The lack of effective analgesic treatment highlights the importance of novel and effective therapeutic strategies. In our laboratory, mesenchymal stromal cell (MSC) based approach showed beneficial immunomodulatory and regenerative effects in a rat model of irreversible radiationinduced colonic ulcers. The goal of my work was to assess the relevance of this model to study radiation-induced visceral persistent hypersensitivity and its modulation by MSC treatment. We first demonstrated that this model is associated with long-lasting visceral hypersensitivity and central neuronal sensitization. In this context we showed then that mast cells (MC) are involved in the mechanism of peripheral sensitization. Moreover, we suggested the implication of the neuro-mediator NO. in the pathophysiology of persistent radiation-induced visceral hypersensitivity. We also suggested that MSC treatment reversed radiation-induced hypersensitivity by a mechanism that in part may involve the modulation of MC activation and/or the decrease in the number of MC and nerve fiber interactions. In addition, MSC treatment reduced the percentage of nitrinergic neurons, increased after irradiation, and restored colonic muscular contractibility. Such processes may promote the therapeutic benefit of MSC observed in our study. In conclusion, this work provided new insights on the therapeutic benefit of MSC in our study model and a new argument in favour of their use in a future clinical trial to cure abdomino-pelvic radiotherapy side effects. (author)

  4. Boron Induces Early Matrix Mineralization via Calcium Deposition and Elevation of Alkaline Phosphatase Activity in Differentiated Rat Bone Marrow Mesenchymal Stem Cells

    Science.gov (United States)

    Movahedi Najafabadi, Bent-al-hoda; Abnosi, Mohammad Hussein

    2016-01-01

    Objective Boron (B) is essential for plant development and might be an essential micronutrient for animals and humans. This study was conducted to characterize the impact of boric acid (BA) on the cellular and molecular nature of differentiated rat bone marrow mesenchymal stem cells (BMSCs). Materials and Methods In this experimental study, BMSCs were extracted and expanded to the 3rdpassage, then cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) complemented with osteogenic media as well as 6 ng/ml and 6 µg/ml of BA. After 5, 10, 15 and 21 days the viability and the level of mineralization was determined using MTT assay and alizarin red respectively. In addition, the morphology, nuclear diameter and cytoplasmic area of the cells were studied with the help of fluorescent dye. The concentration of calcium, activity of alanine transaminase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) as well as sodium and potassium levels were also evaluated using commercial kits and a flame photometer respectively. Results Although 6 µg/ml of BA was found to be toxic, a concentration of 6 ng/ml increased the osteogenic ability of the cell significantly throughout the treatment. In addition it was observed that B treatment caused the early induction of matrix mineralization compared to controls. Conclusion Although more investigation is required, we suggest the prescription of a very low concentration of B in the form of BA or foods containing BA, in groups at high risk of osteoporosis or in the case of bone fracture. PMID:27054120

  5. Transplantation of Mesenchymal Stem Cells for Acute Spinal Cord Injury in Rats: Comparative Study between Intralesional Injection and Scaffold Based Transplantation.

    Science.gov (United States)

    Kim, Yoon Chung; Kim, Young Hoon; Kim, Jang Woon; Ha, Kee Yong

    2016-09-01

    Experimental stem cell therapy for spinal cord injury (SCI) has been extensively investigated. The selection of effective cell transplantation route is also an important issue. Although various types of scaffold have been widely tried as a carrier of stem cells to the injured spinal cord, there was little comparative study to investigate the efficacy of transplantation comparing with conventional transplantation route. A total of 48 Sprague-Dawley rats were subjected to standardized SCI, followed by transplantation of allogeneic mesenchymal stem cells (MSCs), either via intralesional injection (IL group), or via the poly (lactic-co-glycolic acid) (PLGA) scaffold (IP group) or chitosan scaffold (IC group). Engraftment and differentiation of the transplanted cells, expression of neurotrophic factors in the injured spinal cord, and functional recovery were compared with those of the control group. The mean numbers of engrafted MSCs in the IL, IP, and IC groups were 20.6 ± 0.7, 25.6 ± 1.7 and 26.7 ± 1.8 cells/high power filed (HPF), respectively. Results showed higher success rate of MSCs engraftment in the scaffold groups compared to the IL group. Expression of neuroprotective growth factors in the SCI lesions showed no significant differences between the IL, IP, and IC groups. The mean Basso, Beattie and Bresnahan locomotor scales at 6 weeks post-transplantation in the IL, IP, IC, and control groups were 7.9 ± 1.1, 7.9 ± 2.1, 8.7 ± 2.1, and 2.9 ± 1.0, respectively. The functional improvement was most excellent in the IC group. The scaffold based MSC transplantation for acute SCI presented the better cell engraftment and neuroprotective effect compared to the intralesional injection transplantation. PMID:27510379

  6. The impact of silica encapsulated cobalt zinc ferrite nanoparticles on DNA, lipids and proteins of rat bone marrow mesenchymal stem cells.

    Science.gov (United States)

    Novotna, Bozena; Turnovcova, Karolina; Veverka, Pavel; Rössner, Pavel; Bagryantseva, Yana; Herynek, Vit; Zvatora, Pavel; Vosmanska, Magda; Klementova, Mariana; Sykova, Eva; Jendelova, Pavla

    2016-08-01

    Nanomaterials are currently the subject of intense research due to their wide variety of potential applications in the biomedical, optical and electronic fields. We prepared and tested cobalt zinc ferrite nanoparticles (Co0.5Zn0.5Fe2O4+γ [CZF-NPs]) encapsulated by amorphous silica in order to find a safe contrast agent and magnetic label for tracking transplanted cells within an organism using magnetic resonance imaging (MRI). Rat mesenchymal stem cells (rMSCs) were labeled for 48 h with a low, medium or high dose of CZF-NPs (0.05; 0.11 or 0.55 mM); silica NPs (Si-NPs; 0.11 mM) served as a positive control. The internalization of NPs into cells was verified by transmission electron microscopy. Biological effects were analyzed at the end of exposure and after an additional 72 h of cell growth without NPs. Compared to untreated cells, Annexin V/Propidium Iodide labeling revealed no significant cytotoxicity for any group of treated cells and only a high dose of CZF-NPs slowed down cell proliferation and induced DNA damage, manifested as a significant increase of DNA-strand breaks and oxidized DNA bases. This was accompanied by high concentrations of 15-F2t-isoprostane and carbonyl groups, demonstrating oxidative injury to lipids and proteins, respectively. No harmful effects were detected in cells exposed to the low dose of CZF-NPs. Nevertheless, the labeled cells still exhibited an adequate relaxation rate for MRI in repeated experiments and ICP-MS confirmed sufficient magnetic label concentrations inside the cells. The results suggest that the silica-coated CZF-NPs, when applied at a non-toxic dose, represent a promising contrast agent for cell labeling. PMID:26581309

  7. Differentiation of Rat Bone Marrow Mesenchymal Stem Cells Into Neuron-Like Cells In Vitro and Co-Cultured with Biological Scaffold as Transplantation Carrier.

    Science.gov (United States)

    Yue, Wei; Yan, Feng; Zhang, Yue-Lin; Liu, Shu-Ling; Hou, Shu-Ping; Mao, Guo-Chao; Liu, Ning; Ji, Yong

    2016-01-01

    BACKGROUND Autograft and allograft transplantation are used to prompt the regeneration of axons after nerve injury. However, the poor self-regeneration caused by the glial scar and growth inhibitory factors after neuronal necrosis limit the efficacy of these methods. The purpose of this study was to develop a new chitosan porous scaffold for cell seeding. MATERIAL AND METHODS The bone marrow mesenchymal stem cells (BMSCs) and tissue-engineered biomaterial scaffold compound were constructed and co-cultured in vitro with the differentiated BMSCs of Wistar rats and chitosan scaffold in a 3D environment. The purity of the third-generation BMSCs culture was identified using flow cytometry and assessment of induced neuronal differentiation. The scaffolds were prepared by the freeze-drying method. The internal structure of scaffolds and the change of cells' growth and morphology were observed under a scanning electron microscope. The proliferation of cells was detected with the MTT method. RESULTS On day 5 there was a significant difference in the absorbance value of the experimental group (0.549±0.0256) and the control group (0.487±0.0357) (P>0.05); but on day 7 there was no significant difference in the proliferation of the experimental group (0.751±0.011) and the control group and (0.78±0.017) (P>0.05). CONCLUSIONS Tissue engineering technology can provide a carrier for cells seeding and is expected to become an effective method for the regeneration and repair of nerve cells. Our study showed that chitosan porous scaffolds can be used for such purposes. PMID:27225035

  8. Acute and subacute chemical-induced lung injuries: HRCT findings

    Energy Technology Data Exchange (ETDEWEB)

    Akira, Masanori, E-mail: Akira@kch.hosp.go.jp [Department of Radiology, National Hospital Organization Kinki-Chuo Chest Medical Center, 1180 Nagasone-cho, Kita-ku, Sakai City, Osaka 591-8555 (Japan); Suganuma, Narufumi [Department of Environmental Medicine, Kochi Medical School (Japan)

    2014-08-15

    Lung injury caused by chemicals includes bronchitis, bronchiolitis, chemical pneumonitis, pulmonary edema, acute respiratory distress syndrome, organizing pneumonia, hypersensitivity pneumonitis, acute eosinophilic pneumonia, and sarcoid-like granulomatous lung disease. Each chemical induces variable pathophysiology and the situation resembles to the drug induced lung disease. The HRCT features are variable and nonspecific, however HRCT may be useful in the evaluation of the lung injuries and so we should know about HRCT features of lung parenchymal abnormalities caused by chemicals.

  9. Approaches to the evaluation of chemical-induced immunotoxicity.

    OpenAIRE

    Krzystyniak, K; Tryphonas, H; Fournier, M

    1995-01-01

    The immune system plays a crucial role in maintaining health; however, accumulating evidence indicates that this system can be the target for immunotoxic effects caused by a variety of chemicals including the environmental pollutants of polychlorinated biphenyls, chlorinated dibenzo-p-dioxins, pesticides, and heavy metals. Adverse chemical-induced immunomodulation, which is studied within the discipline of immunotoxicology, may be expressed either as immunosuppression/immunodepression or immu...

  10. Acute and subacute chemical-induced lung injuries: HRCT findings

    International Nuclear Information System (INIS)

    Lung injury caused by chemicals includes bronchitis, bronchiolitis, chemical pneumonitis, pulmonary edema, acute respiratory distress syndrome, organizing pneumonia, hypersensitivity pneumonitis, acute eosinophilic pneumonia, and sarcoid-like granulomatous lung disease. Each chemical induces variable pathophysiology and the situation resembles to the drug induced lung disease. The HRCT features are variable and nonspecific, however HRCT may be useful in the evaluation of the lung injuries and so we should know about HRCT features of lung parenchymal abnormalities caused by chemicals

  11. Promise(s of mesenchymal stem cells as an in vitro model system to depict pre-diabetic/diabetic milieu in WNIN/GR-Ob mutant rats.

    Directory of Open Access Journals (Sweden)

    Soundarya L Madhira

    Full Text Available BACKGROUND: Development of model systems have helped to a large extent, in bridging gap to understand the mechanism(s of disease including diabetes. Interestingly, WNIN/GR-Ob rats (Mutants, established at National Centre for Laboratory Animals (NCLAS of National Institute of Nutrition (NIN, form a suitable model system to study obesity with Type 2 diabetes (T2D demonstrating several secondary complications (cataract, cardiovascular complications, infertility, nephropathy etc. The present study has been carried out to explore the potent application(s of multipotent stem cells such as bone marrow mesenchymal stem cells (BM-MSCs, to portray features of pre-diabetic/T2D vis-à-vis featuring obesity, with impaired glucose tolerance (IGT, hyperinsulinemia (HI and insulin resistance (IR seen with Mutant rats akin to human situation. METHODOLOGY/PRINCIPAL FINDINGS: Primary cultures of BM-MSCs (third passage from Mutants, its lean littermate (Lean and parental control (Control were characterized for: proliferation markers, disease memory to mark obesity/T2D/HI/IR which included phased gene expression studies for adipogenic/pancreatic lineages, inflammatory markers and differentiation ability to form mature adipocytes/Insulin-like cellular aggregates (ILCAs. The data showed that BM-MSCs from Mutant demonstrated a state of disease memory, depicted by an upregulated expression of inflammatory markers (IL-6, TNFα; increased stem cell recruitment (Oct-4, Sox-2 and proliferation rates (CD90+/CD29+, PDA, 'S' phase of cell cycle by FACS and BrdU incorporation; accelerated preadipocyte induction (Dact-1, PPARγ2 with a quantitative increase in mature adipocyte formation (Leptin; ILCAs, which were non-responsive to high glucose did confer the Obese/T2D memory in Mutants. Further, these observations were in compliance with the anthropometric data. CONCLUSIONS: Given the ease of accessibility and availability of MSCs, the present study form the basis to report for

  12. Electro-acupuncture promotes survival, differentiation of the bone marrow mesenchymal stem cells as well as functional recovery in the spinal cord-transected rats

    OpenAIRE

    Li Yan; Zhang Yu-Jiao; Li Wen-Jie; Zhang Yan-Qing; Ruan Jing-Wen; Yan Qing; Ding Ying; Dong Hongxin; Zeng Yuan-Shan

    2009-01-01

    Abstract Background Bone marrow mesenchymal stem cells (MSCs) are one of the potential tools for treatment of the spinal cord injury; however, the survival and differentiation of MSCs in an injured spinal cord still need to be improved. In the present study, we investigated whether Governor Vessel electro-acupuncture (EA) could efficiently promote bone marrow mesenchymal stem cells (MSCs) survival and differentiation, axonal regeneration and finally, functional recovery in the transected spin...

  13. Propofol promotes spinal cord injury repair by bone marrow mesenchymal stem cell transplantation

    Institute of Scientific and Technical Information of China (English)

    Ya-jing Zhou; Jian-min Liu; Shu-ming Wei; Yun-hao Zhang; Zhen-hua Qu; Shu-bo Chen

    2015-01-01

    Propofol is a neuroprotective anesthetic. Whether propofol can promote spinal cord injury repair by bone marrow mesenchymal stem cells remains poorly understood. We used rats to investigate spinal cord injury repair using bone marrow mesenchymal stem cell transplantation combined with propofol administrationvia the tail vein. Rat spinal cord injury was clearly alleviated; a large number of newborn non-myelinated and myelinated nerve ifbers appeared in the spinal cord, the numbers of CM-Dil-labeled bone marrow mesenchymal stem cells and lfuorogold-labeled nerve ifbers were increased and hindlimb motor function of spinal cord-injured rats was mark-edly improved. These improvements were more prominent in rats subjected to bone marrow mesenchymal cell transplantation combined with propofol administration than in rats receiving monotherapy. These results indicate that propofol can enhance the therapeutic effects of bone marrow mesenchymal stem cell transplantation on spinal cord injury in rats.

  14. Combined transplantation of bone marrow mesenchymal stem cells and pedicled greater omentum promotes locomotor function and regeneration of axons after spinal cord injury in rats

    Institute of Scientific and Technical Information of China (English)

    Liang Li; Zhiying Zhang; Haiyan Lin; Congli Ren; Chuansen Zhang

    2008-01-01

    BACKGROUND: According to previous studies, the neuroprotective effect of the pedicled greater omentum may be attributed to the secretion of neurotrophic factors and stimulation of angiogenesis. The neurotrophic factors released from the pedicled greater omentum, such as brain-derived neurotrophic factor and neurotrophin 3/4/5 could exert a neuroprotective effect on the damaged host neural and glial cells, and also could induce the transdifferentiation of transplanted bone marrow mesenchymal stem cells (BMSCs) into neural cells. OBJECTIVE: Based on the functions of the omentum of neuro-protection and vascularization, we hypothesize that the transplantation of BMSCs and pedicled greater omentum into injured rat spinal cord might improve the survival rate and neural differentiation of transplanted BMSCs and consequently gain a better functional outcome. DESIGN, TIME AND SETTING: A randomized, controlled animal experiment. The experiments were carried out at the Department of Anatomy, the Secondary Military Medical University of Chinese PLA between June 2005 and June 2007.MATERIALS: Fifteen male inbred Wistar rats, weighing (200±20) g, provided by the Experimental Animal Center of the Secondary Military Medical University of Chinese PLA were used and met the animal ethical standards. Mouse anti-BrdU and mouse anti-NF200 monoclonal antibody were purchased from Boster, China. METHODS: Cell culture: We used inbred Sprague-Dawley rats to harvest bone marrow for culture of BMSCs and transplantation to avoid possible immune rejection. BMSCs were cultured via total bone marrow adherence. Experimental grouping and intervention: The rats were randomly divided into a control group, cell group and combined group, five rats per group. Rats in the control group underwent spinal cord injury (SCI) only, during which an artery clamp with pressure force of 30 g was employed to compress the spinal cord at the T10 level for 30 seconds to produce the SCI model. 5 μL PBS containing 105

  15. Retinal incorporation and differentiation of mesenchymal stem cells intravitreally injected in the injured retina of rats Incorporação e diferenciação retiniana de células tronco mesenquimais intravítreas em ratos

    OpenAIRE

    Paula Castanheira; Leonardo Torquetti; Marcio Bittar Nehemy; Alfredo Miranda Goes

    2008-01-01

    PURPOSE: To evaluate the pattern of retinal integration and differentiation of mesenchymal stem cells (MSCs) injected into the vitreous cavity of rat eyes with retinal injury. METHODS: Adult rat retinas were submitted to laser damage followed by transplantation of DAPI-labeled BM-MSCs grafts. To assess the integration and differentiation of BM-MSCs in laser-injured retina, host retinas were evaluated 2.4 and 8 weeks after injury/transplantation. RESULTS: Our results demonstrated that the graf...

  16. Keratin expression in chemically induced mouse lung adenomas.

    OpenAIRE

    Gunning, W T; Goldblatt, P. J.; Stoner, G D

    1992-01-01

    Chemically induced mouse lung tumors exhibit distinctive growth patterns, characterized by an alveolar or solid appearance, a papillary appearance, or a combination of the two. Lung tumors induced in strain A/J mice by either benzo(a)pyrene (BP) or by N-nitrosoethylurea (ENU) were examined for expression of low- and high-molecular-weight cytokeratins. Simple cytokeratins (low molecular weight) were found in all epithelial cells of the normal mouse lung and in all tumor types, whereas higher-m...

  17. A model for chemically-induced mechanical loading on MEMS

    DEFF Research Database (Denmark)

    Amiot, Fabien

    2007-01-01

    The development of full displacement field measurements as an alternative to the optical lever technique to measure the mechanical response for microelectro-mechanical systems components in their environment calls for a modeling of chemically-induced mechanical fields (stress, strain, and...... displacements). As these phenomena usually arise from species adsorption, adsorbate modification or surface reconstruction, they are surface-related by nature and thus require some dedicated mechanical modeling. The accompanying mechanical modeling proposed herein is intended to represent the chemical part of...... drawn from the energy balance in the accompanying model, highlighting the role of surface functionalization parameters in micromechanical sensors engineering....

  18. Comparison of Human Sodium/Iodide Symporter (hNIS) Gene Expressions between Lentiviral and Adenoviral Vectors in Rat Mesenchymal Stem Cells

    International Nuclear Information System (INIS)

    Quantitative comparison of transgene expression within stem cells between lentivirus and adenovirusmediated delivery systems has not been reported. Here, we evaluated the human sodium iodide symporter (hNIS) gene expression in rat mesenchymal stem cell (rMSC) transduced by lentivirus or adenovirus, and compared the hNIS expression quantitatively between the two delivery systems. Lentiviral-mediated hNIS expressing rMSC (lenti-hNIS-rMSC) was constructed by cloning hNIS gene into pLenti6/UbC/V5-DEST (Invitrogen) to obtain pLenti-hNIS, transducing rMSC with the pLenti-hNIS, and selecting with blasticidin for 3 weeks. Recombinant adenovirus expressing hNIS gene (Rad-hNIS) was produced by homologous recombination and transduction efficiency of Rad-hNIS into rMSC evaluated by Rad-GFP was 19.1±4.7%, 54.0±6.4%, 85.7±8.7%, and 98.4±1.3% at MOI 1, 5, 20, and 100, respectively. The hNIS expressions in lenti-hNIS-rMSC or adeno-hNIS-rMSC were assessed by immunocytochemistry, western blot, and I-125 uptake. Immunocytochemistry and western blot analyses revealed that hNIS expressions in lenti-hNIS-rMSC were greater than those in adeno-hNIS-rMSC at MOI 20 but lower than at MOI 50. However in vitro I-125 uptake test demonstrated that iodide uptake in lenti-hNIS-rMSC (29,704±6,659 picomole/106 cells) was greater than that in adeno-hNIS-rMSC at MOI 100 (6,168±2,134 picomole/106 cells). Despite lower amount of expressed protein, hNIS function in rMSC was greater by lentivirus than by adenovirus mediated expression. Stem cell tracking using hNIS as a reporter gene should be conducted in consideration of relative vector efficiency for transgene expression

  19. Comparison of human sodium iodide symporter (hNIS) gene expression between lentiviral and adenoviral vectors in rat mesenchymal stem cell

    International Nuclear Information System (INIS)

    Quantitative comparison of transgene expression within stem cells between lentivirus and adenovirus-mediated delivery systems has not been done. Here, we evaluated the human sodium iodide symporter (hNIS) gene expression in rat mesenchymal stem cell (rMSC) transduced by lentivirus or adenovirus, and compared the hNIS expression quantitatively between the two delivery systems. Lentiviral-mediated stably hNIS expressing rMSC (lenti-hNIS-rMSC) was constructed by cloning the hNIS gene into pLenti6/UbC/V5-DEST (Invitrogen) to obtain pLenti-hNIS, transducing rMSC with the pLenti-hNIS, and selecting with blasticidin for 3 weeks. Recombinant adenovirus expressing hNIS gene (Rad-hNIS) was produced by homologous recombination and Rad-hNIS transduced rMSC (adeno-hNIS-rMSC) was evaluated for the hNIS expression 48 hours post infection at MOI 1, 5, 20, 50, and 100. The hNIS expression in lenti-hNIS-rMSC or adeno-hNIS-rMSC was assessed by immunocytochemistry, western blot, and I-125 uptake. Immunocytochemistry using mono-clonal anti-hNIS antibody revealed that intensity of hNIS immunoreactivity in lenti-hNIS-rMSC was greater than that in adeno-hNIS-rMSC at MOl 20 but lower than that at MOl 50. Western blot analysis also showed that lenti-hNIS-rMSC was intermediate between adeno-hNIS-rMSCs at MOl 20 and 50 in hNIS expression. However in vitro I-125 uptake test demonstrated that iodide uptake in lenti-hNIS-rMSC (297046659 picomole/106 cells) was greater than that in adeno-hNIS-rMSC at MOI 100 (61682134 picomole/106 cells). These results suggest that lentivirus mediated hNIS expression is greater in terms of hNIS function but lower in terms of hNIS protein amount than adenovirus mediated hNIS expression 48 hours post infection. Stem cell tracking using hNIS as a reporter gene should be conducted in consideration of relative viral efficiency of transgene expression

  20. Comparison of Human Sodium/Iodide Symporter (hNIS) Gene Expressions between Lentiviral and Adenoviral Vectors in Rat Mesenchymal Stem Cells

    Energy Technology Data Exchange (ETDEWEB)

    Park, So Yeon; Lee, Won Woo; Kim, Hyun Joo; Chung, June Key; Kim, Sang Eun [Seoul National University College of Medicine, Seoul (Korea, Republic of); Kim, Sung Jin; Lee, Heui Ran [Medical Research Center, Seoul National University, Seoul (Korea, Republic of)

    2008-10-15

    Quantitative comparison of transgene expression within stem cells between lentivirus and adenovirusmediated delivery systems has not been reported. Here, we evaluated the human sodium iodide symporter (hNIS) gene expression in rat mesenchymal stem cell (rMSC) transduced by lentivirus or adenovirus, and compared the hNIS expression quantitatively between the two delivery systems. Lentiviral-mediated hNIS expressing rMSC (lenti-hNIS-rMSC) was constructed by cloning hNIS gene into pLenti6/UbC/V5-DEST (Invitrogen) to obtain pLenti-hNIS, transducing rMSC with the pLenti-hNIS, and selecting with blasticidin for 3 weeks. Recombinant adenovirus expressing hNIS gene (Rad-hNIS) was produced by homologous recombination and transduction efficiency of Rad-hNIS into rMSC evaluated by Rad-GFP was 19.1{+-}4.7%, 54.0{+-}6.4%, 85.7{+-}8.7%, and 98.4{+-}1.3% at MOI 1, 5, 20, and 100, respectively. The hNIS expressions in lenti-hNIS-rMSC or adeno-hNIS-rMSC were assessed by immunocytochemistry, western blot, and I-125 uptake. Immunocytochemistry and western blot analyses revealed that hNIS expressions in lenti-hNIS-rMSC were greater than those in adeno-hNIS-rMSC at MOI 20 but lower than at MOI 50. However in vitro I-125 uptake test demonstrated that iodide uptake in lenti-hNIS-rMSC (29,704{+-}6,659 picomole/10{sup 6} cells) was greater than that in adeno-hNIS-rMSC at MOI 100 (6,168{+-}2,134 picomole/10{sup 6} cells). Despite lower amount of expressed protein, hNIS function in rMSC was greater by lentivirus than by adenovirus mediated expression. Stem cell tracking using hNIS as a reporter gene should be conducted in consideration of relative vector efficiency for transgene expression.

  1. Improved dopamine transporter binding activity after bone marrow mesenchymal stem cell transplantation in a rat model of Parkinson's disease: small animal positron emission tomography study with F-18 FP-CIT

    International Nuclear Information System (INIS)

    We evaluated the effects of bone marrow-derived mesenchymal stem cells (BMSCs) in a model of Parkinson's disease (PD) using serial F-18 fluoropropylcarbomethoxyiodophenylnortropane (FP-CIT) PET. Hemiparkinsonian rats were treated with intravenously injected BMSCs, and animals without stem cell therapy were used as the controls. Serial FP-CIT PET was performed after therapy. The ratio of FP-CIT uptake in the lesion side to uptake in the normal side was measured. The changes in FP-CIT uptake were also analyzed using SPM. Behavioural and histological changes were observed using the rotational test and tyrosine hydroxylase (TH)-reactive cells. FP-CIT uptake ratio was significantly different in the BMSCs treated group (n = 28) at each time point. In contrast, there was no difference in the ratio in control rats (n = 25) at any time point. SPM analysis also revealed that dopamine transporter binding activity was enhanced in the right basal ganglia area in only the BMSC therapy group. In addition, rats that received BMSC therapy also exhibited significantly improved rotational behaviour and preservation of TH-positive neurons compared to controls. The therapeutic effect of intravenously injected BMSCs in a rat model of PD was confirmed by dopamine transporter PET imaging, rotational functional studies, and histopathological evaluation. (orig.)

  2. Improved dopamine transporter binding activity after bone marrow mesenchymal stem cell transplantation in a rat model of Parkinson's disease: small animal positron emission tomography study with F-18 FP-CIT

    Energy Technology Data Exchange (ETDEWEB)

    Park, Bok-Nam; Lee, Kwanjae; An, Young-Sil [School of Medicine, Ajou University, Department of Nuclear Medicine and Molecular Imaging, Woncheon-dong, Yeongtong-gu, Gyeonggi-do, Suwon (Korea, Republic of); Kim, Jang-Hee; Park, So Hyun [Ajou University School of Medicine, Department of Pathology, Suwon (Korea, Republic of)

    2015-05-01

    We evaluated the effects of bone marrow-derived mesenchymal stem cells (BMSCs) in a model of Parkinson's disease (PD) using serial F-18 fluoropropylcarbomethoxyiodophenylnortropane (FP-CIT) PET. Hemiparkinsonian rats were treated with intravenously injected BMSCs, and animals without stem cell therapy were used as the controls. Serial FP-CIT PET was performed after therapy. The ratio of FP-CIT uptake in the lesion side to uptake in the normal side was measured. The changes in FP-CIT uptake were also analyzed using SPM. Behavioural and histological changes were observed using the rotational test and tyrosine hydroxylase (TH)-reactive cells. FP-CIT uptake ratio was significantly different in the BMSCs treated group (n = 28) at each time point. In contrast, there was no difference in the ratio in control rats (n = 25) at any time point. SPM analysis also revealed that dopamine transporter binding activity was enhanced in the right basal ganglia area in only the BMSC therapy group. In addition, rats that received BMSC therapy also exhibited significantly improved rotational behaviour and preservation of TH-positive neurons compared to controls. The therapeutic effect of intravenously injected BMSCs in a rat model of PD was confirmed by dopamine transporter PET imaging, rotational functional studies, and histopathological evaluation. (orig.)

  3. Neuro-immune interactions in chemical-induced airway hyperreactivity.

    Science.gov (United States)

    Devos, Fien C; Boonen, Brett; Alpizar, Yeranddy A; Maes, Tania; Hox, Valérie; Seys, Sven; Pollaris, Lore; Liston, Adrian; Nemery, Benoit; Talavera, Karel; Hoet, Peter H M; Vanoirbeek, Jeroen A J

    2016-08-01

    Asthma may be induced by chemical sensitisers, via mechanisms that are still poorly understood. This type of asthma is characterised by airway hyperreactivity (AHR) and little airway inflammation. Since potent chemical sensitisers, such as toluene-2,4-diisocyanate (TDI), are also sensory irritants, it is suggested that chemical-induced asthma relies on neuro-immune mechanisms.We investigated the involvement of transient receptor potential channels (TRP) A1 and V1, major chemosensors in the airways, and mast cells, known for their ability to communicate with sensory nerves, in chemical-induced AHR.In vitro intracellular calcium imaging and patch-clamp recordings in TRPA1- and TRPV1-expressing Chinese hamster ovarian cells showed that TDI activates murine TRPA1, but not TRPV1. Using an in vivo model, in which an airway challenge with TDI induces AHR in TDI-sensitised C57Bl/6 mice, we demonstrated that AHR does not develop, despite successful sensitisation, in Trpa1 and Trpv1 knockout mice, and wild-type mice pretreated with a TRPA1 blocker or a substance P receptor antagonist. TDI-induced AHR was also abolished in mast cell deficient Kit(Wsh) (/Wsh) mice, and in wild-type mice pretreated with the mast cell stabiliser ketotifen, without changes in immunological parameters.These data demonstrate that TRPA1, TRPV1 and mast cells play an indispensable role in the development of TDI-elicited AHR. PMID:27126687

  4. 脐带间充质干细胞移植治疗大鼠糖尿病%Umbilical cord mesenchymal stem cell transplantation for the treatment of diabetes mellitus in rats

    Institute of Scientific and Technical Information of China (English)

    李丽; 赵博

    2014-01-01

    背景:脐血间充质干细胞具有很强的增殖能力和分化能力,在趋向分化作用下可以分化成胰岛β细胞,进而起到治疗糖尿病的作用。  目的:观察移植脐带间充质干细胞对大鼠糖尿病的治疗效果。  方法:30只雄性SD大鼠中随机取6只作为对照组,注射生理盐水;其中24只按45 mg/kg的剂量注射链脲霉素建立糖尿病模型后,随机等分为移植组和糖尿病组,移植组大鼠尾静脉注射移植脐带间充质干细胞。  结果与结论:造模后30 d,糖尿病组大鼠空腹血糖维持在较高水平,且高于对照组(P 0.05),而糖尿病组大鼠空腹血糖维持较高水平,且体质量持续下降。提示脐带间充质干细胞移植能有效治疗大鼠糖尿病。%BACKGROUND:Umbilical cord mesenchymal stem cels have strong proliferation and differentiation capacities, and can be induced to differentiate into pancreatic β cels, thereby playing a therapeutic effect on diabetes mel itus. OBJECTIVE:To study the therapeutic effects of transplantation of umbilical cord mesenchymal stem cels for treatment of diabetes melitus in rats. METHODS: Thirty male Sprague-Dawley rats were randomly divided into control group (n=6), transplantation group (n=12) and diabetic group (n=12). Rats in the control group were given normal saline injection. Rats in the other two groups were injected with streptozotocin at a dose of 45 mg/kg to establish diabetic models. After modeling, transplantation of umbilical cord mesenchymal stem celsviatail vein was given in the transplantation group. RESULTS AND CONCLUSION:Thirty days after modeling, the fasting blood glucose was maintained at a higher level in comparison with the control group (P 0.05), but in the diabetic group, the fasting blood glucose level was stil higher and the body mass continued to decrease. These findings indicate that the transplantation of umbilical cord mesenchymal stem cels can be effective

  5. Quantum measurement corrections to chemically induced dynamic nuclear polarization

    CERN Document Server

    Kominis, I K

    2013-01-01

    Chemically induced dynamic nuclear polarization has emerged as a universal signature of spin order in photosynthetic reaction centers. Such polarization, significantly enhanced above thermal equilibrium, is known to result from the nuclear spin sorting inherent in the radical pair mechanism underlying long-lived charge-separated states in photosynthetic reaction centers. We will here show that the recently understood fundamental quantum dynamics of radical-ion-pair reactions open up a new and completely unexpected venue towards obtaining CIDNP signals. The fundamental decoherence mechanism inherent in the recombination process of radical pairs is shown to produce nuclear spin polarizations on the order of $10^4$ times or more higher than thermal equilibrium values at low fields relevant to natural photosynthesis in earth's magnetic field. This opens up the possibility of a fundamentally new exploration of the biological significance of high nuclear polarizations in photosynthesis.

  6. Evaluation of early healing events around mesenchymal stem cell-seeded collagen-glycosaminoglycan scaffold. An experimental study in Wistar rats.

    LENUS (Irish Health Repository)

    Alhag, Mohamed

    2011-03-01

    Tissue engineering using cell-seeded biodegradable scaffolds offers a new bone regenerative approach that might circumvent many of the limitations of current therapeutic modalities. The aim of this experiment was to study the early healing events around mesenchymal stem cell-seeded collagen-glycosaminoglycan scaffolds.

  7. Nuclear factor erythroid-2 related factor 2 overexpressed mesenchymal stem cells transplantation, improves renal function, decreases injuries markers and increases repair markers in glycerol-induced Acute kidney injury rats

    Science.gov (United States)

    Zhaleh, Fateme; Amiri, Fatemeh; Mohammadzadeh-Vardin, Mohammad; Bahadori, Marzie; Harati, Mitra Dehghan; Roudkenar, Mehryar Habibi; Saki, Sasan

    2016-01-01

    Objective(s): Recently cell therapy is a promising therapeutic modality for many types of disease including acute kidney injury (AKI). Due to the unique biological properties, mesenchymal stem cells (MSCs) are attractive cells in this regard. This study aims to transplant MSCs equipped with nuclear factor E2-related factor 2 (Nrf2) in rat experimental models of acute kidney and evaluate regeneration potential of injured kidney especially expression of injury and repaired biomarkers. Materials and methods: Nrf2 was overexpressed in bone marrow-derived MSCs by pcDNA.3.1 plasmid. AKI was induced using glycerol in rat models. The regenerative potential of Nrf2-overexpressed MSCs was evaluated in AKI-Induced animal models using biochemical and histological methods after transplantation. Expression of repaired genes, AQP1 and CK-18, as well as injury markers, Kim-1 and Cystatin C, was also assayed in engrafted kidney sections. Results: Our results revealed that transplantation of Nrf2-overexpressed MSCs into AKI-induced rats decreased blood urea nitrogen and creatinine and ameliorated kidney regeneration throughout 14 days. Upregulation of repaired markers and downregulation of injury markers were considerable 14 days after transplantation. Conclusions: Overexpression of Nrf2 in MSCs suggests a new strategy to increase efficiency of MSC-based cell therapy in AKI. PMID:27114803

  8. To assess the reparative ability of differentiated mesenchymal stem cells in a rat critical size bone repair defect model using high frequency co-registered photoacoustic/ultrasound imaging and micro computed tomography

    Science.gov (United States)

    Zafar, Haroon; Gaynard, Sean; O'Flatharta, Cathal; Doroshenkova, Tatiana; Devine, Declan; Sharif, Faisal; Barry, Frank; Hayes, Jessica; Murphy, Mary; Leahy, Martin J.

    2016-03-01

    Stem cell based treatments hold great potential and promise to address many unmet clinical needs. The importance of non-invasive imaging techniques to monitor transplanted stem cells qualitatively and quantitatively is crucial. The objective of this study was to create a critical size bone defect in the rat femur and then assess the ability of the differentiated mesenchymal stem cells (MSCs) to repair the defect using high frequency co-registered photoacoustic(PA)/ultrasound(US) imaging and micro computed tomography (μCT) over an 8 week period. Combined PA and US imaging was performed using 256 elements, 21 MHz frequency linear-array transducer combined with multichannel collecting system. In vivo 3D PA and US images of the defect bone in the rat femur were acquired after 4 and 8 weeks of the surgery. 3D co-registered structural such as microvasculature and the functional images such as total concentration of haemoglobin (HbT) and the haemoglobin oxygen saturation (sO2) were obtained using PA and US imaging. Bone formation was assessed after 4 and 8 weeks of the surgery by μCT. High frequency linear-array based coregistered PA/US imaging has been found promising in terms of non-invasiveness, sensitivity, adaptability, high spatial and temporal resolution at sufficient depths for the assessment of the reparative ability of MSCs in a rat critical size bone repair defect model.

  9. A Meta-Analysis of the Motion Function through the Therapy of Spinal Cord Injury with Intravenous Transplantation of Bone Marrow Mesenchymal Stem Cells in Rats

    OpenAIRE

    Duo Zhang; Xijing He

    2014-01-01

    Background To compare the efficacy of the therapy of spinal cord injury with intravenous transplantation of bone marrow mesenchymal stem cells (BMSCs) by Meta-analysis. Methods Studies of the BBB scores after intravenous transplantation of BMSCs were searched out from Pubmed, SCI, Cochrane Library, Chinese journal full-text database, China Biology Medicinedisc and Wanfang data-base and analyzed by Review Manager 5.2.5. Results Nine randomized controlled animal trials were selected with 235 ra...

  10. Conditioned Medium from Bone marrow-derived Mesenchymal Stem Cells improves recovery after Spinal Cord Injury in rats: an original strategy to avoid cell transplantation.

    OpenAIRE

    Dorothée Cantinieaux; Renaud Quertainmont; Silvia Blacher; Loïc Rossi; Thomas Wanet; Agnès Noël; Gary Brook; Jean Schoenen; Rachelle Franzen

    2013-01-01

    Spinal cord injury triggers irreversible loss of motor and sensory functions. Numerous strategies aiming at repairing the injured spinal cord have been studied. Among them, the use of bone marrow-derived mesenchymal stem cells (BMSCs) is promising. Indeed, these cells possess interesting properties to modulate CNS environment and allow axon regeneration and functional recovery. Unfortunately, BMSC survival and differentiation within the host spinal cord remain poor, and these cells have been ...

  11. Mesenchymal Stem Cell Attenuates Neutrophil-predominant Inflammation and Acute Lung Injury in an In Vivo Rat Model of Ventilator-induced Lung Injury

    OpenAIRE

    Tian-Shun Lai; Zhi-Hong Wang; Shao-Xi Cai

    2015-01-01

    Background: Subsequent neutrophil (polymorphonuclear neutrophil [PMN])-predominant inflammatory response is a predominant feature of ventilator-induced lung injury (VILI), and mesenchymal stem cell (MSC) can improve mice survival model of endotoxin-induced acute lung injury, reduce lung impairs, and enhance the repair of VILI. However, whether MSC could attenuate PMN-predominant inflammatory in the VILI is still unknown. This study aimed to test whether MSC intervention could attenuate the PM...

  12. Polyurethane/polylactide-based biomaterials combined with rat olfactory bulb-derived glial cells and adipose-derived mesenchymal stromal cells for neural regenerative medicine applications

    International Nuclear Information System (INIS)

    Research concerning the elaboration and application of biomaterial which may support the nerve tissue regeneration is currently one of the most promising directions. Biocompatible polymer devices are noteworthy group among the numerous types of potentially attractive biomaterials for regenerative medicine application. Polylactides and polyurethanes may be utilized for developing devices for supporting the nerve regeneration, like nerve guide conduits or bridges connecting the endings of broken nerve tracts. Moreover, the combination of these biomaterial devices with regenerative cell populations, like stem or precursor cells should significantly improve the final therapeutic effect. Therefore, the composition and structure of final device should support the proper adhesion and growth of cells destined for clinical application. In current research, the three polymer mats elaborated for connecting the broken nerve tracts, made from polylactide, polyurethane and their blend were evaluated both for physical properties and in vitro, using the olfactory-bulb glial cells and mesenchymal stem cells. The evaluation of Young's modulus, wettability and roughness of obtained materials showed the differences between analyzed samples. The analysis of cell adhesion, proliferation and morphology showed that the polyurethane–polylactide blend was the most neutral for cells in culture, while in the pure polymer samples there were significant alterations observed. Our results indicated that polyurethane–polylactide blend is an optimal composition for culturing and delivery of glial and mesenchymal stem cells. - Highlights: • Polyurethane–polylactide blends exhibit different characteristics from pure polymers. • Pure PU and PLA negatively influence on morphology of glial and mesenchymal cells. • PU/PLA blend was neutral for glial and mesenchymal cell proliferation and morphology

  13. Human umbilical cord blood mononuclear cells and chorionic plate-derived mesenchymal stem cells promote axon survival in a rat model of optic nerve crush injury

    OpenAIRE

    CHUNG, SOKJOONG; RHO, SEUNGSOO; KIM, GIJIN; Kim, So-Ra; Baek, Kwang-Hyun; KANG, MYUNGSEO; Lew, Helen

    2016-01-01

    The use of mesenchymal stem cells (MSCs) in cell therapy in regenerative medicine has great potential, particularly in the treatment of nerve injury. Umbilical cord blood (UCB) reportedly contains stem cells, which have been widely used as a hematopoietic source and may have therapeutic potential for neurological impairment. Although ongoing research is dedicated to the management of traumatic optic nerve injury using various measures, novel therapeutic strategies based on the complex underly...

  14. Foxp3-modified bone marrow mesenchymal stem cells promotes liver allograft tolerance through the generation of regulatory T cells in rats

    OpenAIRE

    QI, HAIZHI; Chen, Guangshun; Huang, Yaxun; Si, Zhongzhou; Li, Jiequn

    2015-01-01

    Background The transcription factor forkhead box P3 (Foxp3) is a master regulatory gene necessary for the development and function of CD4+CD25+ regulatory T cells (Tregs). Mesenchymal stem cells (MSC) have recently emerged as promising candidates for cell-based immunosuppression/tolerance induction protocols. Thus, we hypothesized that MSC-based Foxp3 gene therapy would improve immunosuppressive capacity of MSC and induce donor-specific allograft tolerance in rat’s liver allograft model. Meth...

  15. Polyurethane/polylactide-based biomaterials combined with rat olfactory bulb-derived glial cells and adipose-derived mesenchymal stromal cells for neural regenerative medicine applications

    Energy Technology Data Exchange (ETDEWEB)

    Grzesiak, Jakub, E-mail: grzesiak.kuba@gmail.com [Electron Microscopy Laboratory, University of Environmental and Life Sciences, Kozuchowska 5b, 51-631 Wroclaw (Poland); Marycz, Krzysztof [Electron Microscopy Laboratory, University of Environmental and Life Sciences, Kozuchowska 5b, 51-631 Wroclaw (Poland); Szarek, Dariusz [Department of Neurosurgery, Lower Silesia Specialist Hospital of T. Marciniak, Emergency Medicine Center, Traugutta 116, 50-420 Wroclaw (Poland); Bednarz, Paulina [State Higher Vocational School in Tarnów, Mickiewicza 8, 33-100 Tarnów (Poland); Laska, Jadwiga [AGH University of Science and Technology, Faculty of Materials Science and Ceramics, Mickiewicza 30, 30-059 Kraków (Poland)

    2015-07-01

    Research concerning the elaboration and application of biomaterial which may support the nerve tissue regeneration is currently one of the most promising directions. Biocompatible polymer devices are noteworthy group among the numerous types of potentially attractive biomaterials for regenerative medicine application. Polylactides and polyurethanes may be utilized for developing devices for supporting the nerve regeneration, like nerve guide conduits or bridges connecting the endings of broken nerve tracts. Moreover, the combination of these biomaterial devices with regenerative cell populations, like stem or precursor cells should significantly improve the final therapeutic effect. Therefore, the composition and structure of final device should support the proper adhesion and growth of cells destined for clinical application. In current research, the three polymer mats elaborated for connecting the broken nerve tracts, made from polylactide, polyurethane and their blend were evaluated both for physical properties and in vitro, using the olfactory-bulb glial cells and mesenchymal stem cells. The evaluation of Young's modulus, wettability and roughness of obtained materials showed the differences between analyzed samples. The analysis of cell adhesion, proliferation and morphology showed that the polyurethane–polylactide blend was the most neutral for cells in culture, while in the pure polymer samples there were significant alterations observed. Our results indicated that polyurethane–polylactide blend is an optimal composition for culturing and delivery of glial and mesenchymal stem cells. - Highlights: • Polyurethane–polylactide blends exhibit different characteristics from pure polymers. • Pure PU and PLA negatively influence on morphology of glial and mesenchymal cells. • PU/PLA blend was neutral for glial and mesenchymal cell proliferation and morphology.

  16. Spirolactone provides protection from renal fibrosis by inhibiting the endothelial–mesenchymal transition in isoprenaline-induced heart failure in rats

    OpenAIRE

    Zhou H; Xi D; Liu J; Zhao J; Chen S; Guo Z

    2016-01-01

    Hao Zhou, Dan Xi, Jichen Liu, Jinjin Zhao, Si Chen, Zhigang Guo Division of Cardiology, Huiqiao Medical Center, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People’s Republic of China Background: Fibrosis results in excessive accumulation of extracellular matrix proteins, collagen component alteration, and abnormalities in structure and is partly derived from a process called the endothelial–mesenchymal transition involving transforming growth factor &bet...

  17. Spirolactone provides protection from renal fibrosis by inhibiting the endothelial–mesenchymal transition in isoprenaline-induced heart failure in rats

    OpenAIRE

    Zhou, Hao

    2016-01-01

    Hao Zhou, Dan Xi, Jichen Liu, Jinjin Zhao, Si Chen, Zhigang Guo Division of Cardiology, Huiqiao Medical Center, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, People’s Republic of China Background: Fibrosis results in excessive accumulation of extracellular matrix proteins, collagen component alteration, and abnormalities in structure and is partly derived from a process called the endothelial–mesenchymal transition involving transforming growth fac...

  18. Chemically induced skin carcinogenesis: Updates in experimental models (Review)

    Science.gov (United States)

    NEAGU, MONICA; CARUNTU, CONSTANTIN; CONSTANTIN, CAROLINA; BODA, DANIEL; ZURAC, SABINA; SPANDIDOS, DEMETRIOS A.; TSATSAKIS, ARISTIDIS M.

    2016-01-01

    Skin cancer is one of the most common malignancies affecting humans worldwide, and its incidence is rapidly increasing. The study of skin carcinogenesis is of major interest for both scientific research and clinical practice and the use of in vivo systems may facilitate the investigation of early alterations in the skin and of the mechanisms involved, and may also lead to the development of novel therapeutic strategies for skin cancer. This review outlines several aspects regarding the skin toxicity testing domain in mouse models of chemically induced skin carcinogenesis. There are important strain differences in view of the histological type, development and clinical evolution of the skin tumor, differences reported decades ago and confirmed by our hands-on experience. Using mouse models in preclinical testing is important due to the fact that, at the molecular level, common mechanisms with human cutaneous tumorigenesis are depicted. These animal models resemble human skin cancer development, in that genetic changes caused by carcinogens and pro-inflammatory cytokines, and simultaneous inflammation sustained by pro-inflammatory cytokines and chemokines favor tumor progression. Drugs and environmental conditions can be tested using these animal models. keeping in mind the differences between human and rodent skin physiology. PMID:26986013

  19. Chemically induced skin carcinogenesis: Updates in experimental models (Review).

    Science.gov (United States)

    Neagu, Monica; Caruntu, Constantin; Constantin, Carolina; Boda, Daniel; Zurac, Sabina; Spandidos, Demetrios A; Tsatsakis, Aristidis M

    2016-05-01

    Skin cancer is one of the most common malignancies affecting humans worldwide, and its incidence is rapidly increasing. The study of skin carcinogenesis is of major interest for both scientific research and clinical practice and the use of in vivo systems may facilitate the investigation of early alterations in the skin and of the mechanisms involved, and may also lead to the development of novel therapeutic strategies for skin cancer. This review outlines several aspects regarding the skin toxicity testing domain in mouse models of chemically induced skin carcinogenesis. There are important strain differences in view of the histological type, development and clinical evolution of the skin tumor, differences reported decades ago and confirmed by our hands‑on experience. Using mouse models in preclinical testing is important due to the fact that, at the molecular level, common mechanisms with human cutaneous tumorigenesis are depicted. These animal models resemble human skin cancer development, in that genetic changes caused by carcinogens and pro‑inflammatory cytokines, and simultaneous inflammation sustained by pro‑inflammatory cytokines and chemokines favor tumor progression. Drugs and environmental conditions can be tested using these animal models. keeping in mind the differences between human and rodent skin physiology. PMID:26986013

  20. Chemically Induced and Light-Independent Cryptochrome Photoreceptor Activation

    Institute of Scientific and Technical Information of China (English)

    Gesa Rosenfeldt; Rafael Mu(n)oz Viana; Henning D.Mootz; Albrecht G.Von Arnim; Alfred Batschauer

    2008-01-01

    The cryptochrome photoreceptors of higher plants are dimeric proteins. Their N-terminal photosensory domain mediates dimerization, and the unique C-terminal extension (CCT) mediates signaling. We made use of the human FK506-binding protein (FKBP) that binds with high affinity to rapamycin or rapamycin analogs (rapalogs). The FKBP-rapamycin complex is recognized by another protein, FRB, thus allowing rapamycin-induced dimerization of two target proteins. Here we demonstrate by bioluminescence resonance energy transfer (BRET) assays the applicability of this regulated dimerization system to plants. Furthermore, we show that fusion proteins consisting of the C-terminal domain of Arabidopsis cryptochrome 2 fused to FKBP and FRB and coexpressed in Arabidopsis cells specifically induce the expression of cryptochrome-controlled reporter and endogenous genes in darkness upon incubation with the rapalog. These results demonstrate that the activation of cryptochrome signal transduction can be chemically induced in a dose-dependent fashion and uncoupled from the light signal, and provide the groundwork for gain-of-function experiments to study specifically the role of photoreceptors in darkness or in signaling cross-talk even under light conditions that activate members of all photoreceptor families.

  1. Bone marrow mesenchymal stem cells repair spinal cord ischemia/reperfusion injury by promoting axonal growth and anti-autophagy

    OpenAIRE

    Yin, Fei; Meng, Chunyang; Lu, Rifeng; Li, Lei; Zhang, Ying; Chen, Hao; Qin, Yonggang; Guo, Li

    2014-01-01

    Bone marrow mesenchymal stem cells can differentiate into neurons and astrocytes after transplantation in the spinal cord of rats with ischemia/reperfusion injury. Although bone marrow mesenchymal stem cells are known to protect against spinal cord ischemia/reperfusion injury through anti-apoptotic effects, the precise mechanisms remain unclear. In the present study, bone marrow mesenchymal stem cells were cultured and proliferated, then transplanted into rats with ischemia/reperfusion injury...

  2. 人脐带间充质干细胞移植对围绝经期大鼠INHB和AMH的影响%The effect of human umbilical cord mesenchymal stem cells transplantation on INHB and AMH in perimenopausal rats.

    Institute of Scientific and Technical Information of China (English)

    胡佳佳; 尹春艳

    2015-01-01

    Objective:To explore the effect of human umbilical cord mesenchymal stem cells ( hUCMSCs) transplantation on INHB and AMH in perimenopausal SPF SD rats. Method:Vaginal exfoliated cells smear screened out of 30 age 13 to 15 months of SPF SD rat models of perimenopausal,were randomly divided into three groups (A,B and C,n=10). Group A re-ceived no treatment. Group B via the tail vein was grafted with concentration of 1×106/ml of human umbilical cord derived mesenchymal stem cells 1ml,every other day for 1 time,a total of 2 times. Group C and B at the same time and method to have a transplantation with an equal a-mount of cell culture fluid. In the model establishment,the day past 7 days after transplantation and the day past 14 days,AMH and INHB levels were detected in rats. Results:The AMH、IN-HB levels showed no statistical significance when the model establishment and the day past 7 days after transplantation(P > 0. 05). On the day past 14 days after transplantation,the AMH and INHB levels of group B were significantly higher than that of group A、C ( P 0. 05). 末次移植后14天,B组的AMH、INHB水平高于A、C组,差异有统计学意义(P<0. 05);移植后,A、C组的AMH、IN-HB水平逐渐下降,移植后第14 天时下降显著,与成模时比较差异有统计学意义( P<0. 05). 结论:hUCMSCs移植可延缓围绝经期SD大鼠AMH、INHB下降水平,提示对延缓围绝经期大鼠卵巢储备功能下降有积极意义.

  3. A high-throughput chemically induced inflammation assay in zebrafish

    Directory of Open Access Journals (Sweden)

    Liebel Urban

    2010-12-01

    Full Text Available Abstract Background Studies on innate immunity have benefited from the introduction of zebrafish as a model system. Transgenic fish expressing fluorescent proteins in leukocyte populations allow direct, quantitative visualization of an inflammatory response in vivo. It has been proposed that this animal model can be used for high-throughput screens aimed at the identification of novel immunomodulatory lead compounds. However, current assays require invasive manipulation of fish individually, thus preventing high-content screening. Results Here we show that specific, noninvasive damage to lateral line neuromast cells can induce a robust acute inflammatory response. Exposure of fish larvae to sublethal concentrations of copper sulfate selectively damages the sensory hair cell population inducing infiltration of leukocytes to neuromasts within 20 minutes. Inflammation can be assayed in real time using transgenic fish expressing fluorescent proteins in leukocytes or by histochemical assays in fixed larvae. We demonstrate the usefulness of this method for chemical and genetic screens to detect the effect of immunomodulatory compounds and mutations affecting the leukocyte response. Moreover, we transformed the assay into a high-throughput screening method by using a customized automated imaging and processing system that quantifies the magnitude of the inflammatory reaction. Conclusions This approach allows rapid screening of thousands of compounds or mutagenized zebrafish for effects on inflammation and enables the identification of novel players in the regulation of innate immunity and potential lead compounds toward new immunomodulatory therapies. We have called this method the chemically induced inflammation assay, or ChIn assay. See Commentary article: http://www.biomedcentral.com/1741-7007/8/148.

  4. Evaluation of engraftment of superparamagnetic iron oxide-labeled mesenchymal stem cells using three-dimensional reconstruction of magnetic resonance imaging in photo thrombotic cerebral infarction models of rats

    Energy Technology Data Exchange (ETDEWEB)

    Shim, Jae Hyun; Kwak, Byung Kook; Jung, Ji Sung; Park, Se Rah [Chung-Ang University College of Medicine, Seoul (Korea, Republic of)

    2015-06-15

    To evaluate engraftment by visualizing the location of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) three-dimensionally in photothrombotic cerebral infarction (PTCI) models of rats. Magnetic resonance imaging (MRI) of an agarose block containing superparamagnetic iron oxide (SPIO)-labeled hBM-MSCs was performed using a 3.0-T MRI, T2-(T2WI), T2{sup *}-(T2{sup *}WI), and susceptibility-weighted images (SWI). PTCI was induced in 6 rats, and 2.5 x 10(5) SPIO-labeled hBM-MSCs were infused through the ipsilateral internal carotid artery (ICA group) or tail vein (IV group). MRI was performed on days 1, 3, 7, and 14 after stem cell injection. Dark signal regions were confirmed using histology. Three-dimensional MRI reconstruction was performed using the clinical workflow solution to evaluate the engraftment of hBM-MSCs. Volumetric analysis of the engraftment was also performed. The volumes of SPIO-labeled hBM-MSCs in the phantom MRI were 129.3, 68.4, and 25.9 microL using SWI, T2{sup *}WI, and T2WI, respectively. SPIO-labeled hBM-MSCs appeared on day 1 after injection, encircling the cerebral infarction from the ventral side. Dark signal regions matched iron positive cells and human origin (positive) cells. The volume of the engraftment was larger in the ICA group on days 1, 3, and 7, after stem cell injection (p < 0.05 on SWI). SWI was the most sensitive MRI pulse sequence (p < 0.05). The volume of infarction decreased until day 14. The engraftment of SPIO-labeled hBM-MSCs can be visualized and evaluated three-dimensionally in PTCI models of rats. The engraftment volume was larger in the ICA group than IV group on early stage within one week.

  5. Evaluation of engraftment of superparamagnetic iron oxide-labeled mesenchymal stem cells using three-dimensional reconstruction of magnetic resonance imaging in photo thrombotic cerebral infarction models of rats

    International Nuclear Information System (INIS)

    To evaluate engraftment by visualizing the location of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) three-dimensionally in photothrombotic cerebral infarction (PTCI) models of rats. Magnetic resonance imaging (MRI) of an agarose block containing superparamagnetic iron oxide (SPIO)-labeled hBM-MSCs was performed using a 3.0-T MRI, T2-(T2WI), T2*-(T2*WI), and susceptibility-weighted images (SWI). PTCI was induced in 6 rats, and 2.5 x 10(5) SPIO-labeled hBM-MSCs were infused through the ipsilateral internal carotid artery (ICA group) or tail vein (IV group). MRI was performed on days 1, 3, 7, and 14 after stem cell injection. Dark signal regions were confirmed using histology. Three-dimensional MRI reconstruction was performed using the clinical workflow solution to evaluate the engraftment of hBM-MSCs. Volumetric analysis of the engraftment was also performed. The volumes of SPIO-labeled hBM-MSCs in the phantom MRI were 129.3, 68.4, and 25.9 microL using SWI, T2*WI, and T2WI, respectively. SPIO-labeled hBM-MSCs appeared on day 1 after injection, encircling the cerebral infarction from the ventral side. Dark signal regions matched iron positive cells and human origin (positive) cells. The volume of the engraftment was larger in the ICA group on days 1, 3, and 7, after stem cell injection (p < 0.05 on SWI). SWI was the most sensitive MRI pulse sequence (p < 0.05). The volume of infarction decreased until day 14. The engraftment of SPIO-labeled hBM-MSCs can be visualized and evaluated three-dimensionally in PTCI models of rats. The engraftment volume was larger in the ICA group than IV group on early stage within one week.

  6. 骨髓间充质干细胞改善糖尿病大鼠勃起功能障碍的研究%Effect of bone marrow mesenchymal stem cells on improving erectile dysfunction in diabetic rats

    Institute of Scientific and Technical Information of China (English)

    聂永华; 肖明朝; 苟欣; 汤为学; 陈力学; 蔡贤福

    2012-01-01

    目的:探讨骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMMSCs)移植到大鼠阴茎海绵体组织后对勃起功能的影响.方法:体外分离培养BMMSCs并利用形态学及流式细胞术进行鉴定证实.用腹腔注射链脲左菌素(Streptozotocin,STZ)建立糖尿病性大鼠模型,并用阿扑吗啡(Apomorphine,APO)筛选出糖尿病勃起功能障碍(Diabetes mellitus induced erectile dysfunction,DMED)大鼠模型,成模后将BMMSCs(2×106个)移植于大鼠阴茎海绵体内.2周后,分别对正常组、糖尿病组及治疗组进行海绵体内压(Intracorporeal pressure,ICP)及平均动脉压(Mean arterial blood pressure,MAP)测定并取大鼠阴茎海绵体组织,荧光显微镜下观察BMMSCs的局部存活情况,苏木素-伊红染色(Hematoxylin and eosin,HE)观察阴茎海绵体组织中血管.结果:荧光显微镜观察提示BMMSCs能在糖尿病大鼠体内存活;勃起功能测定结果表明治疗组大鼠ICP/MAP比值明显高于糖尿病组;HE染色结果显示在治疗组大鼠阴茎海绵体中血管数目[(12.75±1.89)根/HP]多于糖尿病组[(8.05±1.43)根/HP].结论:BMMSCs能够有效改善糖尿病性大鼠勃起功能.%Objective:To study the effect of bone marrow mesenchymal stem cells(BMMSCs) transplanted into corpus cavernosum on improving erectile dysfunction in diabetic rats. Methods: BMMSCs were separated and cultivated in vitro,then were identified by morphological method and flow cytometry. The diabetic rat model was constructed by intrapertoneal injection of streptozotocin(STZ), then model of diabetes mellitus induced erectile dysfunction(DMED) was sieved by apomorphine(APO). BMMSCs(2×l06 pieces) were transplanted into the corpus cavernosum of the rats in DMED model. Two weeks later.the intracorporeal pressure(ICP) and mean arterial blood pressure(MAP) of normal group,diabetic group and treatment group were determined respectively,the surviving and differentiation of BMMSCs in corpus cavernosum was

  7. Electrophysiological functional recovery in a rat model of spinal cord hemisection injury following bone marrow-derived mesenchymal stem cell transplantation under hypothermia★

    OpenAIRE

    Wang, Dong; Zhang, Jianjun

    2012-01-01

    Following successful establishment of a rat model of spinal cord hemisection injury by resecting right spinal cord tissues, bone marrow stem cells were transplanted into the spinal cord lesions via the caudal vein while maintaining rectal temperature at 34 ± 0.5°C for 6 hours (mild hypothermia). Hematoxylin-eosin staining showed that astrocytes gathered around the injury site and formed scars at 4 weeks post-transplantation. Compared with rats transplanted with bone marrow stem cells under no...

  8. Human umbilical cord blood mesenchymal stem cell transplantation for the treatment of hepatic cirrhosis in rats%人脐血间充质干细胞移植治疗大鼠肝硬化模型*

    Institute of Scientific and Technical Information of China (English)

    詹三华; 张鲁峰; 姚卫民; 杨明辉; 宋晓玲; 焦红亮

    2013-01-01

    BACKGROUND:Studies have shown that cord blood mesenchymal stem cel s can be induced into hepatocyte-like cel s in a specific environment, and the transplanted cel s can exercise the normal function of liver cel s. OBJECTIVE:To establish the rat model of hepatic cirrhosis induced with carbon tetrachloride, and to observe the therapeutic effect of umbilical cord blood mesenchymal stem cel transplantation on a rat model of hepatic cirrhosis. METHODS:Hepatic cirrhosis models were established through carbon tetrachloride inducing, then 20 rat models of hepatic cirrhosis were randomly divided into two groups. The control group was injected with 0.5 mL normal saline via the tail vein;the treatment group was injected with chloromethyl benzamide labeled umbilical cord blood mesenchymal stem cel suspension, and the number of the stem cel s was 1×106. After 2 weeks, al animals were sacrificed. Liver tissues were obtained for the histological observation and the venous blood was col ected to detected the liver function indicators. RESULTS AND CONCLUSION:The liver cel s of the hepatic cirrhosis models were loose and cloudy swel ing, part of the cel s were degenerated and necrosis, the structure of the hepatic lobule was vague, and several pseudolobules with different sizes were formed which meet the diagnostic criteria of hepatic cirrhosis. Compared with the control group, the level of serum albumin in the treatment group was significantly increased (P<0.05), while the levels of bilirubin and transaminases in the treatment group were significantly decreased (P<0.05). There were lots of red chloromethyl benzamide labeled umbilical cord blood mesenchymal stem cel s in the liver tissue of the treatment group. Human umbilical cord blood mesenchymal stem cel s can effectively improve the physiological function of hepatic cirrhosis through tail vein.%  背景:研究表明,脐血间充质干细胞可在特定环境下诱导为肝样细胞,移植入体内能行使正常肝

  9. Characterization of chemically induced liver injuries using gene co-expression modules.

    Directory of Open Access Journals (Sweden)

    Gregory J Tawa

    Full Text Available Liver injuries due to ingestion or exposure to chemicals and industrial toxicants pose a serious health risk that may be hard to assess due to a lack of non-invasive diagnostic tests. Mapping chemical injuries to organ-specific damage and clinical outcomes via biomarkers or biomarker panels will provide the foundation for highly specific and robust diagnostic tests. Here, we have used DrugMatrix, a toxicogenomics database containing organ-specific gene expression data matched to dose-dependent chemical exposures and adverse clinical pathology assessments in Sprague Dawley rats, to identify groups of co-expressed genes (modules specific to injury endpoints in the liver. We identified 78 such gene co-expression modules associated with 25 diverse injury endpoints categorized from clinical pathology, organ weight changes, and histopathology. Using gene expression data associated with an injury condition, we showed that these modules exhibited different patterns of activation characteristic of each injury. We further showed that specific module genes mapped to 1 known biochemical pathways associated with liver injuries and 2 clinically used diagnostic tests for liver fibrosis. As such, the gene modules have characteristics of both generalized and specific toxic response pathways. Using these results, we proposed three gene signature sets characteristic of liver fibrosis, steatosis, and general liver injury based on genes from the co-expression modules. Out of all 92 identified genes, 18 (20% genes have well-documented relationships with liver disease, whereas the rest are novel and have not previously been associated with liver disease. In conclusion, identifying gene co-expression modules associated with chemically induced liver injuries aids in generating testable hypotheses and has the potential to identify putative biomarkers of adverse health effects.

  10. Allelic Variants in Arhgef11 via the Rho-Rock Pathway Are Linked to Epithelial-Mesenchymal Transition and Contributes to Kidney Injury in the Dahl Salt-Sensitive Rat.

    Directory of Open Access Journals (Sweden)

    Zhen Jia

    Full Text Available Previously, genetic analyses identified that variants in Arhgef11 may influence kidney injury in the Dahl salt-sensitive (S rat, a model of hypertensive chronic kidney disease. To understand the potential mechanism by which altered expression and/or protein differences in Arhgef11 could play a role in kidney injury, stably transduced Arhgef11 knockdown cell lines as well as primary cultures of proximal tubule cells were studied. Genetic knockdown of Arhgef11 in HEK293 and NRK resulted in reduced RhoA activity, decreased activation of Rho-ROCK pathway, and less stress fiber formation versus control, similar to what was observed by pharmacological inhibition (fasudil. Primary proximal tubule cells (PTC cultured from the S exhibited increased expression of Arhgef11, increased RhoA activity, and up regulation of Rho-ROCK signaling compared to control (small congenic. The cells were also more prone (versus control to TGFβ-1 induced epithelial-mesenchymal transition (EMT, a hallmark feature of the development of renal interstitial fibrosis, and characterized by development of spindle shape morphology, gene expression changes in EMT markers (Col1a3, Mmp9, Bmp7, and Ocln and increased expression of N-Cadherin and Vimentin. S derived PTC demonstrated a decreased ability to uptake FITC-albumin compared to the small congenic in vitro, which was confirmed by assessment of albumin re-uptake in vivo by infusion of FITC-albumin and immunofluorescence imaging. In summary, these studies suggest that genetic variants in the S form of Arhgef11 via increased expression and/or protein activity play a role in promoting kidney injury in the S rat through changes in cell morphology (Rho-Rock and/or EMT that impact the function of tubule cells.

  11. The anti-oxidative role of micro-vesicles derived from human Wharton-Jelly mesenchymal stromal cells through NOX2/gp91(phox suppression in alleviating renal ischemia-reperfusion injury in rats.

    Directory of Open Access Journals (Sweden)

    Guangyuan Zhang

    Full Text Available Oxidative stress is known as one of the main contributors in renal ischemia/reperfusion injury (IRI. Here we hypothesized that Micro-vesicles (MVs derived from human Wharton Jelly mesenchymal stromal cells (hWJMSCs could protect kidney against IRI through mitigating oxidative stress. MVs isolated from hWJMSCs conditioned medium were injected intravenously in rats immediately after unilateral kidney ischemia for 60 min. The animals were sacrificed at 24 h, 48 h and 2 weeks respectively after reperfusion. Our results show that the expression of NOX2 and reactive oxygen species (ROS in injured kidney tissues was declined and the oxidative stress was alleviated in MVs group at 24 h and 48 h in parallel with the reduced apoptosis and enhanced proliferation of cells. IRI-initiated fibrosis was abrogated by MVs coincident with renal function amelioration at 2 weeks. NOX2 was also found down-regulated by MVs both in human umbilical vein endothelial cells (HUVEC and NRK-52E cell line under hypoxia injury model in vitro. In conclusion, a single administration of hWJMSC-MVs might protect the kidney by alleviation of the oxidative stress in the early stage of kidney IRI through suppressing NOX2 expression. Moreover, it could reduce the fibrosis and improved renal function.

  12. Propofol promotes spinal cord injury repair by bone marrow mesenchymal stem cell transplantation

    OpenAIRE

    Ya-jing Zhou; Jian-min Liu; Shu-ming Wei; Yun-hao Zhang; Zhen-hua Qu; Shu-bo Chen

    2015-01-01

    Propofol is a neuroprotective anesthetic. Whether propofol can promote spinal cord injury repair by bone marrow mesenchymal stem cells remains poorly understood. We used rats to investigate spinal cord injury repair using bone marrow mesenchymal stem cell transplantation combined with propofol administration via the tail vein. Rat spinal cord injury was clearly alleviated; a large number of newborn non-myelinated and myelinated nerve fibers appeared in the spinal cord, the numbers of CM-Dil-l...

  13. Therapeutic effect of mesenchymal stem cell on diabetes In rats%骨髓间充质干细胞对糖尿病大鼠疗效观察

    Institute of Scientific and Technical Information of China (English)

    杨柏梁; 郭丽; 任淑萍; 张桂英; 李鹏; 肖博晗; 刘雅娟

    2009-01-01

    目的 体外诱导骨髓问充质干细胞(mesenchymal stem cells,MSCs)分化为胰岛样细胞,用于干预治疗糖尿病大鼠.方法 取周龄大鼠骨髓细胞,进行MSC培养纯化扩增,用bFGF、胰岛素等培养体系诱导MSC向胰岛细胞转化;双硫腙染色鉴定胰岛样细胞.使用链脲菌素(Streptozotocin,STZ)制作糖尿病大鼠模型,将诱导分化的胰岛样细胞移植至糖尿病模型大鼠,观察干预治疗效果.结果 体外诱导MSCs由长梭形变成多角形并逐渐聚集成圆形,双硫腙染色后细胞团成亮红色,表明为胰岛样细胞.糖尿病模型大鼠移植诱导分化胰岛样细胞1周后,多饮、多尿症状减轻,10 d后,血糖值由(21.352±3.150)mmol/L降低为(14.604±3.236)mmol/L,与对照组(22.256±4.051)mmol/L比较,差异无统计学意义(P>0.05);尿糖值由(18.00±1.00)g/L降低为(8.50±0.25)g/L,与对照组(20.00±0.18)g/L比较,差异有统计学意义(P 0. 05). The urine glucose value decreased (8. 50±0. 25 g/L) and was significantly different from that of control group (20.00±0. 18g/L, P < 0. 05). Conclusion Bone marrow mesenchymaL stem cells could be induced into islet-like cells, and the blood and urine glucose values were decreased after the differentiated cells were transplanted into the diabetes-model rats.

  14. Chemopreventive and chemotherapeutic effect of dietary supplementation of vitamin D on cholangiocarcinoma in a Chemical-Induced animal model.

    Science.gov (United States)

    Chiang, Kun-Chun; Yeh, Chun-Nan; Lin, Kun-Ju; Su, Li-Jen; Yen, Tzu-Chen; Pang, Jong-Hwei S; Kittaka, Atsushi; Sun, Chi-Chin; Chen, Miin-Fu; Jan, Yi-Yin; Chen, Tai C; Juang, Horng-Heng; Yeh, Ta-Sen

    2014-06-15

    Intrahepatic cholangiocarcinoma (ICC) is an aggressive cancer. Vitamin D, a pro-hormone, is getting popular due to its hormone-like functions after converted to its active form, 1α,25(OH)2D3. Here, we show that dietary supplementation with 6 IU/g of vitamin D greatly suppressed ICC initiation and progression without apparent toxicity in a chemically induced rat model. Microarray analysis of rat ICC tissues showed vitamin D supplementation modulated the expressions of several unique genes, including lipocalin 2 (Lcn2), confirmed by RT-qPCR and immunohistochemical (IHC) staining. Further, 53 of 80 human ICC specimens (66%) exhibited high LCN2 expression and LCN2 knockdown in SNU308 cells decreased cell growth and migration, suggesting LCN2 be an oncogene in human ICC. As human ICC SNU1079 cells were treated by 1α,25(OH)2D3, LCN2 expression and cell proliferation were attenuated. The downregulation of LCN2 expression was blunted when vitamin D receptor (VDR) was knocked down, implicating that the in vivo Lcn2 downregulation is a direct consequence of vitamin D supplementation Our results support the prevailing concept that vitamin D status is negatively associated with cancer incidence and mortality and suggest LCN2 may be a potential target against ICC. Further studies of application of vitamin D or its analog against ICC are warranted. PMID:24939880

  15. Distribution of transplanted human mesenchymal stem cells from Wharton’s Jelly in the central nervous systems of the EAE rats

    Directory of Open Access Journals (Sweden)

    Kovalchuk M. V.

    2015-10-01

    Full Text Available Human Wharton’s Jelly MSCs (hWJ-MSCs have a considerable advantage and potential in treating the central nervous system diseases and can be a new alternative treatment of Multiple Sclerosis (MS. Aim. To study the persistence and distribution of hWJ-MSCs along the neuraxis following transplantation in central nervous system of rats with experimental autoimmune encephalomyelitis (EAE, the animal model of MS. Methods. Isolation and cultivation of hWJ-MSCs in vitro. Immunological phenotyping by flow cytometry. EAE induction. Intrathecal (suboccipital injection of MSCs into CNS of SCH-induced EAE rats. Persistence of hWJ-MSCs in the CNS of hWJ-MSCs -treated rats was detected through detection of the human alpha-satellite DNA in the tissue sections and the cerebrospinal fluid (CSF by PCR at days 2, 3, 4 and 5 Results. PCR-assays for alpha-satellite sequences revealed that Human DNA was detected during 5 days following intrathecal injection at the peak of disease in the treated rats. It has been demonstrated that the human DNA was traced in CSF and various segments of a spinal cord. Conclusions. The data obtained suggest that intrathecally delivered hWJ-MSCs, with time, can migrate through the CSF from the injection site to various segments of CNS and persist therein during the first week of post transplantation, which was performed at the EAE disease peak in the xenogeneic setting without immunosuppression. hWJ-MSCs may be considered as a delivery cell source of therapeutic molecules for CNS inflammatory diseases.

  16. Promise(s) of Mesenchymal Stem Cells as an In Vitro Model System to Depict Pre-Diabetic/Diabetic Milieu in WNIN/GR-Ob Mutant Rats

    OpenAIRE

    Madhira, Soundarya L.; Challa, Satya S.; Maniprabha Chalasani; Giridharan Nappanveethl; Ramesh R. Bhonde; Rajanna Ajumeera; Vijayalakshmi Venkatesan

    2012-01-01

    BACKGROUND: Development of model systems have helped to a large extent, in bridging gap to understand the mechanism(s) of disease including diabetes. Interestingly, WNIN/GR-Ob rats (Mutants), established at National Centre for Laboratory Animals (NCLAS) of National Institute of Nutrition (NIN), form a suitable model system to study obesity with Type 2 diabetes (T2D) demonstrating several secondary complications (cataract, cardiovascular complications, infertility, nephropathy etc). The presen...

  17. CD-REST: a system for extracting chemical-induced disease relation in literature.

    Science.gov (United States)

    Xu, Jun; Wu, Yonghui; Zhang, Yaoyun; Wang, Jingqi; Lee, Hee-Jin; Xu, Hua

    2016-01-01

    Mining chemical-induced disease relations embedded in the vast biomedical literature could facilitate a wide range of computational biomedical applications, such as pharmacovigilance. The BioCreative V organized a Chemical Disease Relation (CDR) Track regarding chemical-induced disease relation extraction from biomedical literature in 2015. We participated in all subtasks of this challenge. In this article, we present our participation system Chemical Disease Relation Extraction SysTem (CD-REST), an end-to-end system for extracting chemical-induced disease relations in biomedical literature. CD-REST consists of two main components: (1) a chemical and disease named entity recognition and normalization module, which employs the Conditional Random Fields algorithm for entity recognition and a Vector Space Model-based approach for normalization; and (2) a relation extraction module that classifies both sentence-level and document-level candidate drug-disease pairs by support vector machines. Our system achieved the best performance on the chemical-induced disease relation extraction subtask in the BioCreative V CDR Track, demonstrating the effectiveness of our proposed machine learning-based approaches for automatic extraction of chemical-induced disease relations in biomedical literature. The CD-REST system provides web services using HTTP POST request. The web services can be accessed fromhttp://clinicalnlptool.com/cdr The online CD-REST demonstration system is available athttp://clinicalnlptool.com/cdr/cdr.html. Database URL:http://clinicalnlptool.com/cdr;http://clinicalnlptool.com/cdr/cdr.html. PMID:27016700

  18. Bone marrow mesenchymal stem cells from diabetic versus normal rats for treatment of myocardial infarction%糖尿病与正常大鼠骨髓间充质干细胞移植治疗心肌梗死的比较★

    Institute of Scientific and Technical Information of China (English)

    杨忠路; 王辉山

    2013-01-01

      背景:目前国内外在干细胞移植治疗心肌梗死的研究中,大多采用正常或者年轻的骨髓间充质干细胞作为移植细胞来源。目的:比较糖尿病和正常大鼠骨髓间充质干细胞移植治疗心肌梗死的疗效差别。方法:无菌条件下获取正常大鼠及糖尿病大鼠的骨髓间充质干细胞,建立大鼠心肌梗死模型并随机分为3组,分别于心肌梗死病灶区注射100μL 含有105-106个 F2代正常大鼠或糖尿病大鼠来源骨髓间充质干细胞的细胞混悬液,空白对照组注射100μL 含体积分数20%胎牛血清的 DMEM。移植后1个月行苏木精-伊红染色检测各组梗死区域心肌组织形态的变化;通过免疫组化方法检测 Bcl-2的表达。结果与结论:通过对细胞形态观察及流式细胞术鉴定,大鼠股骨骨髓贴壁培养可获取纯度较高的骨髓间充质干细胞,绘制细胞生长曲线结果显示正常大鼠骨髓间充质干细胞较糖尿病大鼠骨髓间充质干细胞生长快。移植后1个月,正常大鼠干细胞移植组梗死区域心肌组织形态较糖尿病大鼠干细胞移植组及空白对照组明显改善,梗死区域心肌组织 Bcl-2的表达也高于其他2组。证实正常大鼠来源骨髓间充质干细胞较糖尿病大鼠骨髓间充质干细胞生长明显加快,且糖尿病使骨髓干细胞移植治疗心肌梗死的疗效降低。%BACKGROUND: Current studies have shown that bone marrow mesenchymal stem cells from normal or young people usual y serve as a source of transplanted cells in stem cel transplantation treatment of myocardial infarction. OBJECTIVE: To compare the therapeutic effects of bone marrow mesenchymal stem cells from diabetic and normal rats on myocardial infarction. METHODS: Under sterile conditions, bone marrow mesenchymal stem cells from normal and diabetic rats were harvested. Then, rat models of myocardial infarction were established and randomly divided

  19. Bone marrow-derived mesenchymal stem cells repaired but did not prevent gentamicin-induced acute kidney injury through paracrine effects in rats.

    Directory of Open Access Journals (Sweden)

    Luciana A Reis

    Full Text Available This study evaluated the effects of bone marrow-derived mesenchymal stem cells (BMSCs or their conditioned medium (CM on the repair and prevention of Acute Kidney Injury (AKI induced by gentamicin (G. Animals received daily injections of G up to 20 days. On the 10(th day, injections of BMSCs, CM, CM+trypsin, CM+RNase or exosome-like microvesicles extracted from the CM were administered. In the prevention groups, the animals received the BMSCs 24 h before or on the 5(th day of G treatment. Creatinine (Cr, urea (U, FENa and cytokines were quantified. The kidneys were evaluated using hematoxylin/eosin staining and immunohystochemistry. The levels of Cr, U and FENa increased during all the periods of G treatment. The BMSC transplantation, its CM or exosome injections inhibited the increase in Cr, U, FENa, necrosis, apoptosis and also increased cell proliferation. The pro-inflammatory cytokines decreased while the anti-inflammatory cytokines increased compared to G. When the CM or its exosomes were incubated with RNase (but not trypsin, these effects were blunted. The Y chromosome was not observed in the 24-h prevention group, but it persisted in the kidney for all of the periods analyzed, suggesting that the injury is necessary for the docking and maintenance of BMSCs in the kidney. In conclusion, the BMSCs and CM minimized the G-induced renal damage through paracrine effects, most likely through the RNA carried by the exosome-like microvesicles. The use of the CM from BMSCs can be a potential therapeutic tool for this type of nephrotoxicity, allowing for the avoidance of cell transplantations.

  20. Profiles of metabolites and gene expression in rats with chemically induced hepatic necrosis

    NARCIS (Netherlands)

    Heijne, W.H.M.; Lamers, R.J.A.N.; Bladeren, P.J. van; Groten, J.P.; Nesselrooij, J.H.J. van; Ommen, B. van

    2005-01-01

    This study investigated whether integrated analysis of transcriptomics and metabolomics data increased the sensitivity of detection and provided new insight in the mechanisms of hepatotoxicity. Metabolite levels in plasma or urine were analyzed in relation to changes in hepatic gene expression in ra

  1. Protective role of lignan-converting bacteria on chemically-induced breast cancer in gnotobiotic rats

    OpenAIRE

    Mabrok, Hoda Hussein Bakr

    2013-01-01

    Enterolignans (enterodiol and enterolactone) exhibit structural similarity to estradiol and have therefore been hypothesized to modulate hormone related cancers such as breast cancer. The bioactivation of the plant lignan secoisolariciresinol diglucoside (SDG) requires the transformation by intestinal bacteria including the deglycosylation of SDG to secoisolariciresinol (SECO) followed by demethylation and dehydroxylation of SECO to enterodiol (ED). Finally, ED is dehydrogenated to enterolact...

  2. Differentiation of Bone Marrow Mesenchymal Stem Cells in Congenital Spina Bifida of Fetus Rats%骨髓间充质干细胞在先天性脊椎裂胎鼠脊髓中的分化

    Institute of Scientific and Technical Information of China (English)

    高飞; 袁正伟

    2012-01-01

    Objective To investigate the bone of survival and differentiation of mesenchymal stem cells ( MSC) in congenital spina bifida of fetus rats and to explore the theoretical clues for cell replacement of neural lube defects. Methods In sterile condition, rat bone marrow was flushed and cells were seeded in culture flask. Passage 3 of MSC were transfected by adenovirus enhanced green fluorescent protein ( ECFP) gene and then were injected into congenital spina bifida of fetus rats through fine glass needle. On 20 days of gestation, fetus rats underwent operation and spinal cords were removed for fixation and dehydration and got frozen sections. Expressions of Nestin( marker of neural stem cells) and glial fibrillary acidic protein(GFAP,marker of neurogliocyte) in transplanted MSC with EGKF were tested by immunofluores-cence. Results MSC surface antigen by flow cytometry showed CD34 ( - ) , CD44 occupying 95% , and CD90 occupying 99%. Transfection efficiency of adenovirus - EGFP to MSC was 61 %. The rate of dominant spina bifida was 64. 3% in teratogenic group. MSC distributed on the surface of spinal cord,or migrated into it. Some MSCs displayed different cell shapes such as long spindle or multiangle. Expressions of Nestin and GFAP were present in transplanted MSC by immunofluorescence assay. Conclusion MSC with EGFP can survive in fetus spinal cord after transplantation and differentiate into neural stem cells and neurogliocytes.%目的 观察大鼠骨髓间充质干细胞(MSC)在先天性脊椎裂胎鼠脊髓中的存活与分化,探寻细胞替代治疗先天性脊椎裂的方法.方法 用贴壁培养法进行大鼠MSC原代培养并传代,携带增强型绿色荧光蛋白(EGFP)基因的腺病毒转染第3代骨髓MSC.将转染的MSC注射到先天性脊椎裂胎鼠的脊髓中.母鼠孕20 d时取先天性脊椎裂胎鼠的脊椎,固定脱水后做冷冻切片,免疫荧光方法检测脊髓中EGFP阳性MSC中巢蛋白(Nestin)、胶质纤维

  3. Amniotic Fluid-Derived Mesenchymal Stem Cells Cut Short the Acuteness of Cisplatin-Induced Nephrotoxicity in Sprague-Dawley Rats

    Science.gov (United States)

    Al-Husseiny, Fatma; Sobh, Mohamed Ahmed; Ashour, Rehab H.; Foud, Samah; Medhat, Tarek; El-Gilany, Abdel-Hady; Elghannam, Doaa; Abdel-Ghaffar, Hassan; Saad, Mohamed-Ahdy; Sobh, Mohamed

    2016-01-01

    Background and objectives Cisplatin is a nephrotoxic chemotherapeutic agent. So, preventive measures worth to be evaluated. Human amniotic fluid stem cells (hAFSCs) in prevention or amelioration of cisplatin-induced acute kidney injury (AKI) in Sprague-Dawley rates have been tested. Methods 80 Sprague-Dawley rats (250~300 g) were used and divided into 4 major groups, 20 rats each. Group I: Saline-injected group. Group II: Cisplatin-injected group (5 mg/kg I.P). Group III: Cisplatin-injected and hAFSCs-treated group (5×106 hAFSCs I.V. one day after cisplatin administration). Group IV: Cisplatin-injected and culture media-treated group. Each major group was further divided into 4 equal subgroups according to the timing of sacrifice; 4, 7, 11 and 30 days post-cisplatin injection. Renal function tests were done. Kidney tissue homogenate oxidative stress parameters malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) were determined. Histopathological scoring systems for active injury, regenerative and chronic changes were analyzed separately. Results hAFSCs characterization and differentiation was proved. Cisplatin injection resulted in a significant increase in serum creatinine and MDA and decrease in SOD, GSH and creatinine clearance. These changes were attenuated early by day 4 with the use of hAFSCs. Cisplatin injection induced tubular necrosis, atrophy, inflammatory cells infiltration and fibrosis. The use of hAFSCs was associated with significantly lowered injury score at day 4, 7, 11 and 30 with marked regenerative changes starting from day 4. Conclusion hAFSCs have both a protective and regenerative activities largely through an antioxidant activity. This activity cut short the acuteness of cisplatin nephrotoxicity. PMID:27426088

  4. Chemical -induced apoptotic cell death in tomato cells : involvement of caspase-like proteases

    NARCIS (Netherlands)

    Jong, de A.J.; Hoeberichts, F.A.; Yakimova, E.T.; Maximova, E.; Woltering, E.J.

    2000-01-01

    A new system to study programmed cell death in plants is described. Tomato (Lycopersicon esculentum Mill.) suspension cells were induced to undergo programmed cell death by treatment with known inducers of apoptosis in mammalian cells. This chemical-induced cell death was accompanied by the characte

  5. Chemically-induced mouse lung tumors: applications to human health assessments [Poster 2014

    Science.gov (United States)

    A state-of-the-science workshop on chemically-induced mouse lung tumors was conducted by U.S. Environmental Protection Agency to discuss issues related to the use of mouse lung tumor data in human health assessments. Naphthalene, styrene, and ethylbenzene were chosen for the anal...

  6. Chemically-induced Mouse Lung Tumors: Applications to Human Health Assessments

    Science.gov (United States)

    A state-of-the-science workshop on chemically-induced mouse lung tumors was conducted by U.S. Environmental Protection Agency to better understand the mouse lung tumor data’s role in human health assessments. Three environmental chemicals - naphthalene, styrene, and ethylbe...

  7. Infusion of mesenchymal stem cells limits fibrogenesis in irradiated rat lung%间充质干细胞对延缓大鼠放射性肺纤维化进展的作用研究

    Institute of Scientific and Technical Information of China (English)

    常鹏宇; 夏诚诚; 侯雪; 石硙岩; 宋宇哲; 张玉宇; 赵钦; 马利新; 曲雅勤

    2015-01-01

    Dawley rats were enrolled in this study.For establishing the model of radiation-induced lung injury,36 rats were randomly selected to administrate a dose of 15 Gy to their whole right lungs.Thereafter,the irradiated rats were randomly classified into three groups.Each group contained 12 rats.Phosphate buffered sodium (PBS group),human skin-derived fibroblasts (fibroblast group) and human adipose tissue-derived mesenchymal stem cells (Ad-MSC group) were injected into rat through the tail vein respectively.The rest 12 rats without semi-thoracic irradiation were set as healthy controls.At 24 weeks post-irradiation,radiological imaging and pathological analyses were used for evaluating the histological changes in irradiated lungs.The pathological analyses included H&E staining for reflecting pulmonary architecture,Masson's Trichrome (M.T) staining for evaluating extracellular matrix (ECM) formation and IHC-staining for identifying the expressions of alpha-smooth muscle actin (α-SMA) and TGF-β1 in irradiated lungs.Additionally,the concentrations of HGF and TGF-β1 in serum and BALF were tested by ELISA assay.Quantitative real-time PCR assay was used to test the gene-expressions in irradiated lungs,including Collagen Ⅰ-α1 and Collagen Ⅲ-α1.Results The chest radiological imaging revealed that high-density shadows were presented in the irradiated lungs at 24 weeks post-irradiation.The lower lobes were predominant disease.However,the lesion of high-density shadow was not obvious in Ad-MSC group.Histological analysis by H&E staining indicated that the irradiated lungs in both PBS and fibroblast groups lost their architectures,representing by alveolar collapse and thickening alveolar septa.Moreover,compared to Ad-MSC group,excessive ECM formation together with significantly up-regulated expressions of Collagen Ⅰ-α1 and Collagen Ⅲ-α1 (F =4.39,7.73,P < 0.05) were also found within the injured tissue treated with PBS or fibroblasts.Apart from this,obvious expressions of

  8. In vitro differentiation of rat mesenchymal stem cells into photoreceptors%大鼠骨髓间充质干细胞体外诱导为光感受器样细胞的研究

    Institute of Scientific and Technical Information of China (English)

    张枝桥; 董方田

    2008-01-01

    Objective To isolate and culture rat mesenchymal stem cells (MSCs) and induce them to differentiate into photoreceptor cells in vitro. Methods It was a experimental study. MSCs were isolated and attached to the wall of culture dishes by their specific adherent ability. Then the cells were characterized by flow cytometry. MSCs were induced by murine EGF for 8-10 days in-vitro. Then we used immunohistochemistry staining and flow cytometry to see whether they could express rhodopsin and differentiated into photoreceptor cells. Results Many MSCs, which were highly purified, were obtained.After induction by EGF, 15.32% MSCs expressed rhodopsin. Conclusion After induction by EGF,approximately 15.32% MSCs could differentiate into photoreceptor cells.%目的 探讨体外培养的大鼠骨髓问允质干细胞(MSC)诱导为光感受器样细胞的可能性.方法 实验研究.采用贴壁筛选法分离、培养SD大鼠骨髓MSC细胞,经流式细胞仪鉴定后,用含小鼠表皮生长因子(EGF)的培养液诱导MSC细胞8~10 d.用免疫组织化学染色观察诱导后的细胞是否表达视紫红质,并用流式细胞术检测表达视紫红质的细胞比例.结果 传3代的MSC细胞生长曲线呈S型,显示培养细胞具有正常的分裂增殖特性;超过70%的传1代和传2代细胞为CD90单阳性,从传3代绌胞开始CD90单阳性的百分数一直高于93%,即采用贴壁筛选法可获得大量高纯度的大鼠骨髓MSC细胞.经EGF诱导后的细胞有15.32%±0.92%表达视紫红质,而未经诱导的细胞表达视紫红质的比例为1.64%±0.78%.结论 经EGF诱导后,15.32%的大鼠MSC可分化为光感受器样细胞.

  9. Chondroitinase ABC plus bone marrow mesenchymal stem cells for repair of spinal cord injury☆

    OpenAIRE

    Zhang, Chun; He, Xijing; Li, Haopeng; Wang, Guoyu

    2013-01-01

    As chondroitinase ABC can improve the hostile microenvironment and cell transplantation is proven to be effective after spinal cord injury, we hypothesized that their combination would be a more effective treatment option. At 5 days after T8 spinal cord crush injury, rats were injected with bone marrow mesenchymal stem cell suspension or chondroitinase ABC 1 mm from the edge of spinal cord damage zone. Chondroitinase ABC was first injected, and bone marrow mesenchymal stem cell suspension was...

  10. Mesenchymal stem cells preconditioned with trimetazidine promote neovascularization of hearts under hypoxia/reoxygenation injury

    OpenAIRE

    Hu, Xiaowu; Yang, Junjie; Wang, Ying; Zhang, You; Ii, Masaaki; Shen, Zhenya; Hui, Jie

    2015-01-01

    Background: Cell-based angiogenesis is a promising treatment for ischemic diseases; however, survival of implanted cells is impaired by the ischemic microenvironment. In this study, mesenchymal stem cells (MSCs) for cell transplantation were preconditioned with trimetazidine (TMZ). We hypothesized that TMZ enhances the survival rate of MSCs under hypoxic stimuli through up-regulation of HIF1-α. Methods and results: Bone marrow-derived rat mesenchymal stem cells were preconditioned with 10 μM ...

  11. Neuroprotection and immunomodulation by xenografted human mesenchymal stem cells following spinal cord ventral root avulsion

    OpenAIRE

    Ribeiro, Thiago B.; Duarte, Adriana S. S.; Longhini, Ana Leda F.; Fernando Pradella; Farias, Alessandro S.; Angela C. M. Luzo; Oliveira, Alexandre L. R.; Sara Teresinha Olalla Saad

    2015-01-01

    The present study investigates the effects of xenotransplantation of Adipose Tissue Mesenchymal Stem Cells (AT-MSCs) in animals after ventral root avulsion. AT-MSC has similar characteristics to bone marrow mesenchymal stem cells (BM-MSCs), such as immunomodulatory properties and expression of neurotrophic factors. In this study, Lewis rats were submitted to surgery for unilateral avulsion of the lumbar ventral roots and received 5 × 105 AT-MSCs via the lateral funiculus. Two weeks after cell...

  12. 骨髓间充质干细胞移植影响慢性脑缺血大鼠认知功能及海马区Nogo-A 、NgR 的表达%Transplantation of bone marrow mesenchymal stem cells improves cognitive function of rats with chronic cerebral ischemia and decreases Nogo-A and NgR protein expression in the hippocampus

    Institute of Scientific and Technical Information of China (English)

    王燕琳; 闫禹; 宋波; 张惠莉; 龚光明; 陈思; 方慧; 许予明

    2012-01-01

    BACKGROUND: Transplantation of bone marrow mesenchymal stem cells can decrease Nogo-A and NgR expression in rats with spinal cord injury and cerebral infarction. OBJECTIVE: To investigate the effects of transplantation of bone marrow mesenchymal stem cells on cognitive function of rats with chronic cerebral ischemia and Nogo-A and NgR protein expression in the hippocampus. METHODS: Thi rty Sprague-Dawley rats were randomly divided into sham-surgery, model and bone marrow mesenchymal stem cells groups. Rats in the latter two groups were established into model of chronic cerebral ischemia by permanent ligation of bilateral common carotid artery. In the bone marrow mesenchymal stem cells group, at 7 days after chronic cerebral ischemia induction, bone marrow mesenchymal stem cells (1×107) isolated by ficoll density gradient centrifugation were transfused into rats via the tailvein. RESULTS AND CONCLUSION: Compared with sham-surgery group, the average escape latency was significantly shortened (P < 0.01), the number that rats passed through the platform was significantly increased (P < 0.05), and Nogo-A and NgR expression in the hippocampal ti ssue was significantly decreased (P < 0.05) in the bone marrow mesenchymal stem cells group. These findings suggest that transplantation of bone marrow mesenchymal stem cells protects spatial learning and memory abilities of rats with chronic cerebral ischemia. This effect is possibly related to decreased Nogo-A and NgR protein expression.%背景:骨髓间充质干细胞移植可降低脊髓损伤及脑梗死大鼠的Nogo-A 及NgR 的表达.目的:观察骨髓间充质干细胞移植对慢性脑缺血大鼠认知功能及海马区Nogo-A 及NgR 蛋白的表达.方法:将30 只SD大鼠随机等分为假手术组、模型组及骨髓间充质干细胞组,后2 组采用双侧颈总动脉永久结扎法建立大鼠慢性脑缺血模型,骨髓间充质干细胞组在建模7 d后尾静脉注射移植用ficoll密度梯度离心法分

  13. Imprint Control of BaTiO3 Thin Films via Chemically Induced Surface Polarization Pinning.

    Science.gov (United States)

    Lee, Hyungwoo; Kim, Tae Heon; Patzner, Jacob J; Lu, Haidong; Lee, Jung-Woo; Zhou, Hua; Chang, Wansoo; Mahanthappa, Mahesh K; Tsymbal, Evgeny Y; Gruverman, Alexei; Eom, Chang-Beom

    2016-04-13

    Surface-adsorbed polar molecules can significantly alter the ferroelectric properties of oxide thin films. Thus, fundamental understanding and controlling the effect of surface adsorbates are crucial for the implementation of ferroelectric thin film devices, such as ferroelectric tunnel junctions. Herein, we report an imprint control of BaTiO3 (BTO) thin films by chemically induced surface polarization pinning in the top few atomic layers of the water-exposed BTO films. Our studies based on synchrotron X-ray scattering and coherent Bragg rod analysis demonstrate that the chemically induced surface polarization is not switchable but reduces the polarization imprint and improves the bistability of ferroelectric phase in BTO tunnel junctions. We conclude that the chemical treatment of ferroelectric thin films with polar molecules may serve as a simple yet powerful strategy to enhance functional properties of ferroelectric tunnel junctions for their practical applications. PMID:26901570

  14. 骨髓间充质干细胞移植治疗脑出血大鼠的实验研究%Experimental study of bone mesenchymal stem cells transplantation for intracerebral hemorrhage in rats

    Institute of Scientific and Technical Information of China (English)

    徐德才; 张少军; 娄飞云; 姜之全; 韩易

    2014-01-01

    目的:探讨骨髓间充质干细胞移植对脑出血大鼠的行为和血肿周围神经细胞凋亡的影响。方法全骨髓贴壁法分离培养大鼠骨髓间充质干细胞,使用立体定向纹状体注入胶原酶法制作大鼠脑出血模型。 SD大鼠30只,随机分为对照组和移植组(各15只),对照组制作脑出血模型,不移植;移植组制作脑出血模型并于造模后48h经立体定向脑内注射1×105个干细胞,并于移植后1d、3d、5d、7d、14d进行神经功能缺损评分,对照组在相应时间点同样评分,各组再根据时间的不同随机分成五个亚组(每组3只),在相应时间点处死大鼠行免疫组织化学法检测细胞凋亡。结果在移植后1 d、3 d,两组大鼠评分及血肿周围脑组织内凋亡细胞的数量均较高,两组之间无显著性差异。在移植后第5 d、7 d,两组大鼠评分及血肿周围脑组织内凋亡细胞的数量开始下降,移植组下降幅度大于对照组,两组之间差异显著。移植后14 d,两组之间无显著性差异。结论骨髓间充质干细胞移植可明显促进大鼠脑出血后神经功能的恢复,其机制可能与下调血肿周围神经细胞凋亡有关。%Objective To explore the effect of the behavioral test and neurological cell apoptosis on transplantation of bone mesenchymal stem cells (BMSCs) for experimental intracerebral hemorrhage (ICH) in rats.Methods BMSCs were isolated and cultured using the whole bone marrow adherent method.A rat model of ICH was established by stereotaxical injection of collagenase Ⅰinto the right caudate nucleus .30 Sprague-Dawley ( SD ) rats were randomly divided into control group ( 15 ) and transplantation group (15).The control group were injected with collagenaseⅠonly,the transplant-ation group were transfused with BMSCs (1 ×105/rat) at 48 hours after surgery .On 1,3,5,7,14 days afer transplantation neurological function tests were performed among

  15. 黄芪多糖诱导大鼠骨髓间充质干细胞分化的特性%Induction of Astragalus Polysaccharides on Differentiation of Rat Bone Marrow ;Mesenchymal Stem Cells

    Institute of Scientific and Technical Information of China (English)

    刘永琦; 李静雅; 蔡玲; 窦娟娟

    2014-01-01

    目的:观察黄芪多糖对大鼠骨髓间充质干细胞(BMSCs)向神经细胞、脂肪细胞、成骨细胞和软骨细胞分化特性的影响,为开发有效而又低毒的分化诱导剂提供依据。方法采用全骨髓贴壁筛选法分离、纯化无特定病原体级Wistar大鼠BMSCs。用噻唑蓝(MTT)法筛选出黄芪多糖的合适浓度,取F3代细胞,采用随机数字表法分为对照组和诱导组(神经诱导、成脂诱导、成骨诱导、软骨诱导),采用甲苯胺蓝染色、油红O染色、茜素红染色检测神经细胞、脂肪细胞、软骨细胞表面特异性标记物。用Western Blot技术检测神经元特异性烯醇化酶(NSE)、脂蛋白酯酶(LPL)、胶原蛋白Ⅰ、胶原蛋白Ⅱ。分析黄芪多糖对F3代BMSCs向神经、脂肪、软骨和骨的诱导分化作用。结果 MTT显示,在1 g/L黄芪多糖诱导48 h下细胞增殖明显,甲苯胺蓝染色阳性,而油红O、茜素红染色阴性。Western Blot法检测显示NSE为阳性表达,LPL、胶原蛋白Ⅰ、胶原蛋白Ⅱ均为阴性表达。结论黄芪多糖可诱导大鼠BMSCs定向分化为神经细胞,而未向脂肪细胞、成骨细胞和软骨细胞分化。%Objective To investigate the effect of astragalus polysaccharides (APS) on the differentiation of rat bone marrow mesenchymal stem cells (BMSCs) to neurones, adipocytes, osteoblasts and chondrocytes, and provide basis for the development of effective and low toxic differentiation inducing agents. Methods BMSCs were isolated from SPF Wistar rats, purified, expanded and cultured to family 3. The appropriate concentration of APS was filtered out by MTT assay. The F3 cells were randomly divided into control group and induced group (neural induction, adipogenic induction, osteogenic induction, cartilage induction). The effects of APS and classical chemical drugs on differentiation were measured by toluidine blue, oil red o and alizarin red staining. The protein expression of NSE, LPL, collagen

  16. SD大鼠骨髓间充质干细胞的分离及5-氮胞苷对其的诱导分化作用%Induction of rat bone marrow mesenchymal stem cells by 5-azacytidine

    Institute of Scientific and Technical Information of China (English)

    邓丽群; 沈法荣; 金红峰; 王欢

    2014-01-01

    Objective To induce rat bone marrow mesenchymal stem cells (SD- MSCs) with 5- azacytidine (5- aza). Methods The primary culture of SD- MSCs was isolated from the bone marrow of SD- rats by the whole bone marrow adhesion separation. The morphology of SD- MSCs was observed with phase contrast microscopy. The cellcycle was analyzed by flow cy-tometry. The multilineage differentiation capability of SD- MSCs was examined by adipogenic differentiation method. SD- MSCs was induced by 5- aza, and the expression of hyperpolarization- activated cyclic nucleotide- gated channel (HCN2) in induced SD- MSCs was examined by immunohistochemistry and Western blot. Results Through isolation and culture, SD- MSCs devel-oped a spindle appearance and had radiated colony growth. Growth curve of SD- MSCs presented S shape. SD- MSCs kept a high potential of multiplication. After adipogenic differentiation, lipid droplets presented red under oil red O staining. Immunohis-tochemistry and Western blot showed that SD- MSCs expressed positive for HCN2 after induced by 5- aza. Conclusion SD- MSCs can be separated, proliferated and purified under the whole bone marrow adherence culture and 5- aza can success-ful y induce differentiation of SD- MSCs.%目的:探讨SD大鼠骨髓间充质干细胞(SD- MSCs)体外分离的方法条件、生物学特性及5-氮胞苷对其的诱导分化作用。方法采用全骨髓贴壁法分离培养SD- MSCs,进行形态学观察,绘制生长曲线,流式细胞仪对其细胞周期进行分析,并通过成脂肪诱导鉴定,给予5-氮胞苷进行诱导分化,通过免疫组化和蛋白免疫印迹法,观察其对超极化激活环核苷酸门控的通道蛋白(HCN2)表达情况。结果 SD- MSCs以长梭形和多角形为主,呈放射状集落生长,细胞生长曲线呈S形,增殖能力比较活跃。成脂肪诱导后,细胞胞质内有大量橙红色脂滴形成。不同浓度的5-氮胞苷对SD- MSCs进行诱导分化后,7.5

  17. Experimental study of rat mesenchymal stem cells transfected with Sonic Hedgehog gene%Shh基因转染大鼠骨髓间充质干细胞的可行性

    Institute of Scientific and Technical Information of China (English)

    吴晓明; 唐滔; 杨进福; 谭志平; 张伟

    2012-01-01

    Objective To evaluate the feasibility of genetic modification of mesenchymal stem cells (MSC) with Sonic Hedgehog(Shh) gene.Methods The pcDNA3.l-Shh eukaryotic expression plasmid was constructed and its correctness evaluated by the restriction enzyme analysis and sequencing. MSC were isolated from Wistar rats by density gradient centrifugation and purified,transfected with pcDNA3.1-Shh,blank plasmid pcDNA3.1 ( - ) or pmaxGFP respectively by NucleofectorTM.The protein expression of Shh in MSC was detected by Western blot after 48 hours.Results Correct construction of pcDNA3.1-Shh was identified by the methods of restriction enzyme analysis and nucleotide sequence determination. The expression of green fluorescent protein (GFP) could be observed by fluorescence microscopy after 48 hours.The expression of Shh gene was detected by Western blot.But the MSC transfected with empty plasmid expression was not detected.Conclusions Recombinant Eukaryotic expression plasmid pcDNA3.1-Shh is successfully detected in rat MSC.It may provide experimental rationales for the future gene therapy.%目的 探讨Sonic Hedgehog(Shh)基因有效转染大鼠骨髓间充质干细胞(BMSC)的可行性.方法 构建pcDNA3.1-Shh真核表达质粒,酶切和测序鉴定.密度梯度离心-贴壁培养法获取Wistar大鼠BMSC,电击穿孔法将pcDNA3.1-Shh、pcDNA3.1(一)空质粒和pmaxGFP报告质粒转染进BMSC,48 h后Western印迹法检测Shh基因的表达情况.结果 酶切和测序结果证实pcDNA3.1-Shh正确性.转染48 h后,荧光显微镜可观察到绿色荧光蛋白(GFP)的表达,有40%以上的细胞发出绿色荧光.Western印迹法检测证实有Shh基因表达,而转染pcDNA3.1(一)空质粒的BBMSC未检测到表达.结论 重组真核表达质粒pcDNA3.1-Shh转染大鼠BMSC后能有效表达Shh基因,为进一步Shh基因与细胞联合治疗大鼠心肌缺血模型提供了实验依据.

  18. Mammalian models of chemically induced primary malignancies exploitable for imaging-based preclinical theragnostic research

    OpenAIRE

    Liu, Yewei; YIN Ting; Feng, Yuanbo; Cona, Marlein Miranda; Huang, Gang; Liu, Jianjun; Song, Shaoli; Jiang, Yansheng; Xia, Qian; Swinnen, Johannes V; Bormans, Guy; Himmelreich, Uwe; Oyen, Raymond; Ni, Yicheng

    2015-01-01

    Compared with transplanted tumor models or genetically engineered cancer models, chemically induced primary malignancies in experimental animals can mimic the clinical cancer progress from the early stage on. Cancer caused by chemical carcinogens generally develops through three phases namely initiation, promotion and progression. Based on different mechanisms, chemical carcinogens can be divided into genotoxic and non-genotoxic ones, or complete and incomplete ones, usually with an organ-spe...

  19. Influence of duration of exposition in the Chernobyl zone on spontaneous and chemically induced mutagenesis

    International Nuclear Information System (INIS)

    The cycle of researches according to influence of an exposition of laboratory mice of line Af in a zone of Chernobyl disaster on spontaneous and chemically induced mutagenesis has been performed. The frequency of micronucleated erythrocytes in the mouse bone marrow was increase in term of an exposition in a zone of Chernobyl disaster with 1 till 4 months in comparison with the control. (authors)

  20. Improving analytical methods for protein-protein interaction through implementation of chemically inducible dimerization

    OpenAIRE

    Tonni Grube Andersen; Nintemann, Sebastian J.; Magdalena Marek; Halkier, Barbara A.; Alexander Schulz; Meike Burow

    2016-01-01

    When investigating interactions between two proteins with complementary reporter tags in yeast two-hybrid or split GFP assays, it remains troublesome to discriminate true- from false-negative results and challenging to compare the level of interaction across experiments. This leads to decreased sensitivity and renders analysis of weak or transient interactions difficult to perform. In this work, we describe the development of reporters that can be chemically induced to dimerize independently ...

  1. Comparison of Skeletal Effects of Ovariectomy Versus Chemically Induced Ovarian Failure in Mice

    OpenAIRE

    Wright, Laura E; Christian, Patricia J.; Rivera, Zelieann; Van Alstine, William G.; L Funk, Janet; L Bouxsein, Mary; Hoyer, Patricia B.

    2008-01-01

    Bone loss associated with menopause leads to an increase in skeletal fragility and fracture risk. Relevant animal models can be useful for evaluating the impact of ovarian failure on bone loss. A chemically induced model of menopause in which mice gradually undergo ovarian failure yet retain residual ovarian tissue has been developed using the chemical 4-vinylcyclohexene diepoxide (VCD). This study was designed to compare skeletal effects of VCD-induced ovarian failure to those associated wit...

  2. Immunosuppressive effects and mechanism of rat bone marrow mesenchymal stem cells on allogeneic T lymphocytes%大鼠骨髓间充质干细胞和免疫调节作用

    Institute of Scientific and Technical Information of China (English)

    高峰; 林雨佳; 李国良; 吴德全

    2011-01-01

    目的 研究一氧化氮(nitric oxide,NO)在骨髓间充质干细胞(mesenchymal stem cells,MSCs)的免疫调节作用.方法 贴壁筛选法分离培养Lewis大鼠骨髓间充质干细胞.以刀豆蛋白A(concanavalin A,ConA)作为刺激因素,SD大鼠外周血分离的T淋巴细胞作为反应细胞,采用CCK-8法检测T淋巴细胞与MSCs共培养后其增殖能力的变化.然后采用RT-PCR、Western blot检测MSCs诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)mRNA与蛋白的表达;Griess法检测上清液亚硝酸盐含量;ELISA检测上清液Th1细胞因子IFN-γ和Th2细胞因子IL-4的水平.结果 MSCs明显抑制ConA诱导的T淋巴细胞增殖,抑制作用因细胞比例的不同而有所区别.当实验组MSC:T为1:10时最明显,增殖率为(33.83±2.10)%,而1:80组[(78.62±3.80)%]与不含MSCs的阳性对照组[(79.03±1.70)%]差异无统计学意义(P>0.05).实验组MSCs的iNOS mRNA与蛋白的表达以及上清液中亚硝酸盐含量均较阳性对照组明显上调,1:10~1:40的范围内呈明显递增趋势,但1:80组与1:40组差异无统计学意义(P>0.05).在加入iNOS特异性抑制剂2-甲基-2-巯基硫酸脲(S-methylisothiourea sulfate,SMT)后,各组T淋巴细胞增殖率基本恢复.实验组IFN-γ含量随MSCs所占比例的降低而增加,各组IL-4含最差异无统计学意义(P>0.05).结论 MSCs对同种异体外周血T淋巴细胞的增殖有免疫调节作用,机制可能与其分泌的可溶性因子NO影响了 Th1/Th2平衡有关.%Objective To observe the role of nitric oxide(NO) in the immune regulatory effect of bone marrow mesenchymal stem cells(MSCs). Methods Bone marrow MSCs were isolated from Lewis rats by adherence screening. Concanavalin A (ConA) was adopted as the stimulator and T-lymphocyte isolated from peripheral blood of SD rats as the reactive cells. The changes of the ability of T-lymphocyte proliferation, when co-cultured with MSCs, were measured by CCK-8 assay. The inducible nitric

  3. Dental mesenchymal stem cells

    OpenAIRE

    Kaukua, Nina

    2014-01-01

    Mesenchymal stem cells have been found in various tissues and act as source for renewal and repair. The mouse incisor tooth continuously grows throughout life, implicating that there are stem cell niches constantly contributing with cells. The composition of these stem cell niches is not fully understood. Here, we show that Schwann cells on the peripheral nerves in the close proximity to the incisor tooth constitute a stem cell niche. Transgenic mouse models were used to label ...

  4. 磁性/荧光量子点双功能纳米粒子标记大鼠骨髓间充质干细胞%Magnetic/luminescent quantum dots bifunctional nanoparticles labeling of rat bone mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    颜荣华; 单鸿; 聂立波; 李丹; 王劲; 朱康顺; 王红飞; 马栋; 张黎明

    2010-01-01

    目的 利用磁性/荧光量子点双功能纳米粒子双标记大鼠骨髓间充质干细胞(BMSC),探讨标记效率和可行性.方法 分离、纯化及培养大鼠BMSC.制备二氧化硅(SiO_2)同时包裹四氧化三铁(Fe_3O_4)和碲化镉(CdTe)荧光量子点的磁性/荧光双功能纳米粒子Fe_3O_4@CdTe@SiO_2:,利用铁浓度为25μg/ml的Fe_3O_4@CdTe@SiO_2标记BMSC,未标记的BMSC作为对照组.通过荧光显微镜观察细胞内纳米粒子的荧光表达情况.双标记后的BMSC分成8个不同数量级(3×10~6~1×10~3个),应用1.5 T磁共振扫描仪进行T_1 WI、T_2 WI和T_2~* WI 3个序列成像.分光光度计测量细胞铁含量,普鲁士蓝染色显示细胞内铁.结果 荧光显微镜下可见双标记组细胞内Fe_3O_4@CdTe@SiO_2:纳米粒子显示为红色荧光,细胞标记率达到90%以上.磁共振成像可见3×10~6~5×10~4双标记组细胞较对照组T_2 WI、T_2~* WI信号减低,两组之间的信号差异随数量级减小而相应减小,T_2~* WI信号变化最明显,T_2WI次之.细胞铁含量为(14.05±4.15)pg/细胞,普鲁士蓝染色可见双标记细胞胞质内蓝染的含铁颗粒.结论 Fe_3O_4@CdTe@SiO_2,纳米粒子可同时对大鼠BMSC进行磁性和荧光双标记,有望成为磁共振和光学成像活体示踪移植后BMSC的有效手段.%Objective To evaluate the dual-labeling efficiency of magnetic and luminescent quantum dots bifunctional nanoparticles to rat bone mesenchymal stem cells ( BMSC). Methods Rat BMSC were isolated, purified, and expanded. Magnetic/luminescent bifunctional nanoparticles (Fe_3O_4@ CdTe@SiO_2) , were prepared by using silicon dioxide(SiO_2) to encapsulate simultaneously Fe_3O_4 and CdTe quantum dots. BMSC were incubated with the Fe_3O_4@ CdTe@ SiO_2 nanoparticles which iron concentration was 25 μg/mL Fluorescence microscope was used to detect the fluorescence of the intracellular nanoparticles. The dual-labeled BMSC with various concentration underwent ex vivo MR imaging

  5. Direct evidence of epithelial-mesenchymal interdependence in situ.

    OpenAIRE

    Adatia, A K

    1980-01-01

    In the basal odontogenic zone of the mandibular incisor of the rat the cytological evidence of the toxicity of 40 mg cyclophosphamide per kg, 24 hours following a single intraperitoneal injection, was confined to the proliferating mesenchyme. Localised loss of precursors of odontoblasts cessation of basal odontogenesis and acellularity of the related part of the pulp ensured. The cells of the internal enamel epithelium bordering this region did not differentiate into ameloblasts and appeared ...

  6. Bone Marrow Mesenchymal Stem Cells Combined with Polylactic Glycolic Acid Scaffold for Rat Treating Bladder Function after Spinal Cord Injury%骨髓间充质干细胞复合聚乳酸-羟基乙酸支架材料移植修复大鼠脊髓损伤后膀胱功能的实验研究

    Institute of Scientific and Technical Information of China (English)

    桂有富; 周兴; 符厚圣; 潘建刚

    2011-01-01

    Objectives To study separately the effect of bone marrow mesenchymal stem cells(BMSCs)and differentiated neural-like cells combined with polylactic glycolic acid(PLGA)scaffolds,both composite which restore to SD rat,s bladder function after spinal cord injury.Methods Bold scaffold,BMSCs-PLGA complex,and neural-like cells-PLGA complex were separately transplanted into SD rat models with the transection for right spinal cord,the control group which is a direct suture half crosscut spinal cord group.And obsvered SD rat models during 60 days after operation.Results BMSCs and neural-like cells can be exactly adhere to,proliferate and grow on the PLGA;BMSCs and neural-like cells combined with PLGA,which can restore to spinal cord and promote the recovery of bladder function of SD rat models.Conclusions BMSCs and differentiated neural-like cells combined with PLGA scaffolds,which can be used as a method of tissue engineering,to treat bladder function caused by spinal cord injury.%目的 研究骨髓间充质干细胞(Bone Marrow Mesenchymal Stem Cells,BMSCs)及其分化生成的神经样细胞分别复合聚乳酸-羟基乙酸(Polylactic Glycolic Acid,PLGA)支架材料移植治疗大鼠脊髓损伤的效果.方法 分别将单纯的PLGA、BMSCs-PLGA复合物以及神经样细胞-PLGA复合物移植入3组半横切的SD大鼠脊髓损伤模型中,对照组为直接缝合脊髓组.术后观察4组动物60d.结果 BMSCs及其分化生成的神经样细胞能在PLGA上良好的黏附、增殖和生长,两者复合移植后能促进脊髓及膀胱排尿功能的恢复.结论 BMSCs及神经样细胞复合PLGA移植可作为组织工程治疗脊髓损伤后膀胱功能的一种方法.

  7. Transplantation of autologous bone marrow-derived mesenchymal stem cells for traumatic brain injury

    Institute of Scientific and Technical Information of China (English)

    Jindou Jiang; Xingyao Bu; Meng Liu; Peixun Cheng

    2012-01-01

    Results from the present study demonstrated that transplantation of autologous bone marrow-derived mesenchymal stem cells into the lesion site in rat brain significantly ameliorated brain tissue pathological changes and brain edema, attenuated glial cell proliferation, and increased brain-derived neurotrophic factor expression. In addition, the number of cells double-labeled for 5-bromodeoxyuridine/glial fibrillary acidic protein and cells expressing nestin increased. Finally, blood vessels were newly generated, and the rats exhibited improved motor and cognitive functions. These results suggested that transplantation of autologous bone marrow-derived mesenchymal stem cells promoted brain remodeling and improved neurological functions following traumatic brain injury.

  8. 大鼠骨髓间充质干细胞定向成骨诱导的动态观察%Directed differentiation of rat bone marrow mesenchymal stem cells into osteoblasts A dynamic observational study

    Institute of Scientific and Technical Information of China (English)

    郭立达; 夏冰; 王捷

    2008-01-01

    背景:骨髓间充质干细胞由于其具有多向分化潜能和高增殖潜能而在组织工程领域具有广泛的应用前景.目的:观察大鼠骨髓间充质干细胞在向成骨细胞分化过程中细胞及生物活性变化.设计、时间和地点:随机对照实验,实验于2004-07/2006-06在解放军广州军区广州总医院医学实验科干细胞组织工程实验室完成.材料:成年清洁级SD大鼠20只,雌雄不拘用于取骨髓.方法:按随机数字表法将动物分为实验组和对照组,每组10只.通过改良的骨髓培养法分离成年大鼠骨髓间充质干细胞,对照组以基础培养液(10%胎牛血清+高糖DMEM+100U/L青霉素+100 U/L链霉素+2 mmol/L谷氨酰胺)培养,实验组用条件培养液(基础培养液,10 mmol/Lβ-甘油磷酸纳,10-7 mol/L地塞米松,50 mg/L维生素C)进行诱导培养.一段时间后制备细胞爬片.主要观察指标:①用倒置显微镜和透射电子显微镜进行细胞形态学观察.②采用Gomori改良钙钴法进行碱性磷酸酶染色,应用免疫细胞化学染色方法测定骨钙素的表达.结果:骨髓间充质干细胞接种48 h后,较多细胞贴壁,72 h后增殖,生长良好;7~9 d时细胞融合率可达到80%.透射电镜下骨髓间充质干细胞的细胞核较大,形态幼稚.经过体外成骨诱导的骨髓间充质干细胞表现出增殖、聚集、结节和矿化期的阶段性形态特征,诱导1周后表达碱性磷酸酶活性,2周后骨钙素表达呈阳性.结论:骨髓间充质干细胞经体外成骨定向诱导1周后进入成骨细胞的转化过程,并可以保持一定程度的增殖活性,可作为骨组织工程的种子细胞.%BACKGROUND:With the potential of multi-directional differentiation and high proliferation, bone marrow mesenchymal stem cells (BMSCs) have wide application prospect in tissue engineering. OBJECTIVE:To investigate changes in cells and bioactivity in the differentiation from BMSCs into osteoblasts. DESIGN, TIME AND

  9. Proteomic Alterations in B Lymphocytes of Sensitized Mice in a Model of Chemical-Induced Asthma

    OpenAIRE

    Steven Haenen; Jeroen A.J. Vanoirbeek; Vanessa De Vooght; Liliane Schoofs; Benoit Nemery; Elke Clynen; Hoet, Peter H. M.

    2015-01-01

    Introduction and Aim The role of B-lymphocytes in chemical-induced asthma is largely unknown. Recent work demonstrated that transferring B lymphocytes from toluene diisocyanate (TDI)-sensitized mice into naïve mice, B cell KO mice and SCID mice, triggered an asthma-like response in these mice after a subsequent TDI-challenge. We applied two-dimensional difference gel electrophoresis (2D-DIGE) to describe the “sensitized signature” of B lymphocytes comparing TDI-sensitized mice with control mi...

  10. Nuclear magnetic resonance in pulse radiolysis. Chemically induced dynamic nuclear polarization

    International Nuclear Information System (INIS)

    Nuclear magnetic resonance and chemically induced dynamic nuclear polarization (CIDNP) were applied to the study of pulse radiolysis. Samples were irradiated with a 3-MeV electron beam from the Argonne Van de Graaff accelerator in an EPR magnet (approximately 4000 G) which had axial holes for beam access. A fast flow system transferred the irradiated solution to the rotating 5-mm NMR sample tube. The NMR spectra of mixtures of sodium acetate and methanol were presented to demonstrate the features of the CIDNP in pulse radiolysis

  11. Rats

    Directory of Open Access Journals (Sweden)

    Alexey Kondrashov

    2012-01-01

    Full Text Available We aimed to perform a chemical analysis of both Alibernet red wine and an alcohol-free Alibernet red wine extract (AWE and to investigate the effects of AWE on nitric oxide and reactive oxygen species production as well as blood pressure development in normotensive Wistar Kyoto (WKY and spontaneously hypertensive rats (SHRs. Total antioxidant capacity together with total phenolic and selected mineral content was measured in wine and AWE. Young 6-week-old male WKY and SHR were treated with AWE (24,2 mg/kg/day for 3 weeks. Total NOS and SOD activities, eNOS and SOD1 protein expressions, and superoxide production were determined in the tissues. Both antioxidant capacity and phenolic content were significantly higher in AWE compared to wine. The AWE increased NOS activity in the left ventricle, aorta, and kidney of SHR, while it did not change NOS activity in WKY rats. Similarly, increased SOD activity in the plasma and left ventricle was observed in SHR only. There were no changes in eNOS and SOD1 expressions. In conclusion, phenolics and minerals included in AWE may contribute directly to increased NOS and SOD activities of SHR. Nevertheless, 3 weeks of AWE treatment failed to affect blood pressure of SHR.

  12. Endothelial and Epithelial Cell Transition to a Mesenchymal Phenotype Was Delineated by Nestin Expression.

    Science.gov (United States)

    Chabot, Andréanne; Hertig, Vanessa; Boscher, Elena; Nguyen, Quang Trinh; Boivin, Benoît; Chebli, Jasmine; Bissonnette, Lyse; Villeneuve, Louis; Brochiero, Emmanuelle; Dupuis, Jocelyn; Calderone, Angelino

    2016-07-01

    Endothelial and epithelial cell transition to a mesenchymal phenotype was identified as cellular paradigms implicated in the appearance of fibroblasts and development of reactive fibrosis in interstitial lung disease. The intermediate filament protein nestin was highly expressed in fibrotic tissue, detected in fibroblasts and participated in proliferation and migration. The present study tested the hypothesis that the transition of endothelial and epithelial cells to a mesenchymal phenotype was delineated by nestin expression. Three weeks following hypobaric hypoxia, adult male Sprague-Dawley rats characterized by alveolar and perivascular lung fibrosis were associated with increased nestin protein and mRNA levels and marked appearance of nestin/collagen type I((+)) -fibroblasts. In the perivascular region of hypobaric hypoxic rats, displaced CD31((+)) -endothelial cells were detected, exhibited a mesenchymal phenotype and co-expressed nestin. Likewise, epithelial cells in the lungs of hypobaric hypoxic rats transitioned to a mesenchymal phenotype distinguished by the co-expression of E-cadherin and collagen. Following the removal of FBS from primary passage rat alveolar epithelial cells, TGF-β1 was detected in the media and a subpopulation acquired a mesenchymal phenotype characterized by E-cadherin downregulation and concomitant induction of collagen and nestin. Bone morphogenic protein-7 treatment of alveolar epithelial cells prevented E-cadherin downregulation, suppressed collagen induction but partially inhibited nestin expression. These data support the premise that the transition of endothelial and epithelial cells to a mesenchymal cell may have contributed in part to the appearance nestin/collagen type I((+)) -fibroblasts and the reactive fibrotic response in the lungs of hypobaric hypoxic rats. J. Cell. Physiol. 231: 1601-1610, 2016. © 2015 Wiley Periodicals, Inc. PMID:26574905

  13. Protective effects of traditional Chinese medicine on mesenchymal stem cells transplanted into the hippocampus of rats with post-traumatic stress disorder%中药干预对创伤性应激障碍大鼠海马区移植间充质干细胞的保护作用***☆

    Institute of Scientific and Technical Information of China (English)

    富文俊; 敖海清; 曾蕾; 徐志伟

    2013-01-01

    BACKGROUND:Transplantation of bone marrow mesenchymal stem cel s shows great potential in repair of hippocampal injury. However, low survival rate of transplanted stem cel s remains unsolved. OBJECTIVE:To summarize the protective effects of traditional Chinese medicine on bone marrow mesenchymal stem cel s transplanted in the hippocampus of rats with post-traumatic stress disorder. METHODS:The databases of Pubmed, and CNKI were retrieved for papers published from January 2000 to January 2012 with key words“mesenchymal stem cel s, transplantation, survival and post-traumatic stress disorder (PTSD), hippocampus”in English and Chinese. A total of 109 papers were retrieved. Among them, 23 papers were included in the final analysis. RESULTS AND CONCLUSION:Post-traumatic stress disorder is a complex disease influenced by biological, psychological, social and other factors and remains unsolved in treatment. Traditional Chinese medicine exhibits a unique advantage in this research field. Hippocampal damage caused by post-traumatic stress disorder can be repaired by transplantation of bone marrow mesenchymal stem cel s. Bone marrow mesenchymal stem cel transplantation combined with intervention of Xiaoyaosan, a traditional Chinese medicine, is expected to increase the survival rate of transplanted stem cel s.%  背景:骨髓间充质干细胞移植修复损伤海马具有较高的研究潜力,但是目前移植后干细胞的成活率较低这一问题尚未得到有效解决。目的:综述中药干预对创伤性应激障碍大鼠海马区移植间充质干细胞的保护作用。方法:应用计算机检索2000年1月至2012年1月PubMed数据库相关文章,检索词为“mesenchymal stem cel s,transplantation,survival,post-traumatic stress disorder (PTSD),hippocampus”,并限定文章语言种类为English,同时检索2000年1月至2012年1月CNKI数据库相关文章,检索词为“间充质干细胞,移植,存活率,创伤后

  14. Cistanche deserticola plus bone marrow mesenchymal stem cell transplantation for treating spinal cord injury in rats%口服肉苁蓉联合骨髓间充质干细胞移植治疗大鼠脊髓损伤

    Institute of Scientific and Technical Information of China (English)

    兰静; 闫金玉; 夏润福; 李剑锋

    2014-01-01

    BACKGROUND:Studies have shown that bone marrow mesenchymal stem cells under certain conditions can differentiate into nerve cells. Bone marrow mesenchymal stem celltransplantation can rebuild nervous system function and improve functional disorders in patients. Glycosides of cistanche also have a protective effect against nerve cellinjury. Their combination has been less reported. OBJECTIVE:To observe the effectiveness of Cistanche deserticola and bone marrow mesenchymal stem celltransplantation on spinal cord injury in rats. METHODS:Fifty adult Wistar rats with spinal cord injury were randomly divided into four group:a Cistanche deserticola group (intragastric administration of 20 mL/kg Cistanche deserticola concentrated solution per day), a celltransplantation group (10μL of 1×108/L bone marrow mesenchymal stem cellsuspension), a combination group (10μL of 1×108/L bone marrow mesenchymal stem cellsuspension+intragastric administration of 20 mL/kg Cistanche deserticola concentrated solution per day) and a control group (intragastric administration of 20 mL/kg normal saline per day). The intragastric administration lasted for 30 days in each group. RESULTS AND CONCLUSION:After 30 days of treatment, the expression of Nestin was significantly higher in the combination group than the other groups. After 12 weeks, Basso, Beattie, and Bresnahan scores was significantly higher in the combination group than the other groups (P<0.05);somatosensory and motor evoked potential latencies were also improve significantly in the combination group compared with the other groups (P<0.05). These findings indicate that oral administration of Cistanche deserticola combined with bone marrow mesenchymal stem cells can significantly improve the motor and neurophysiological function of spinal cord injury rats. Cistanche deserticola can improve the survival of transplanted bone marrow mesenchymal stem cells in rats with spinal cord injury.%背景:研究表明骨髓间充质干细胞

  15. Mesenchymal stem cells.

    Science.gov (United States)

    Ding, Dah-Ching; Shyu, Woei-Cherng; Lin, Shinn-Zong

    2011-01-01

    Stem cells have two features: the ability to differentiate along different lineages and the ability of self-renewal. Two major types of stem cells have been described, namely, embryonic stem cells and adult stem cells. Embryonic stem cells (ESC) are obtained from the inner cell mass of the blastocyst and are associated with tumorigenesis, and the use of human ESCs involves ethical and legal considerations. The use of adult mesenchymal stem cells is less problematic with regard to these issues. Mesenchymal stem cells (MSCs) are stromal cells that have the ability to self-renew and also exhibit multilineage differentiation. MSCs can be isolated from a variety of tissues, such as umbilical cord, endometrial polyps, menses blood, bone marrow, adipose tissue, etc. This is because the ease of harvest and quantity obtained make these sources most practical for experimental and possible clinical applications. Recently, MSCs have been found in new sources, such as menstrual blood and endometrium. There are likely more sources of MSCs waiting to be discovered, and MSCs may be a good candidate for future experimental or clinical applications. One of the major challenges is to elucidate the mechanisms of differentiation, mobilization, and homing of MSCs, which are highly complex. The multipotent properties of MSCs make them an attractive choice for possible development of clinical applications. Future studies should explore the role of MSCs in differentiation, transplantation, and immune response in various diseases. PMID:21396235

  16. 12 hours after cerebral ischemia is the optimal time for bone marrow mesenchymal stem cell transplantation

    Institute of Scientific and Technical Information of China (English)

    Seyed Mojtaba Hosseini; Mohammad Farahmandnia; Zahra Razi; Somayeh Delavarifar; Benafsheh Shakibajahromi

    2015-01-01

    Cell therapy using stem cell transplantation against cerebral ischemia has been reported. However, it remains controversial regarding the optimal time for cell transplantation and the transplantation route. Rat models of cerebral ischemia were established by occlusion of the middle cerebral artery. At 1, 12 hours, 1, 3, 5 and 7 days after cerebral ischemia, bone marrow mesenchymal stem cells were injected via the tail vein. At 28 days after cerebral ischemia, rat neurological function was evaluated using a 6-point grading scale and the pathological change of ischemic cerebral tissue was observed by hematoxylin-eosin staining. Under the lfuorescence microscope, the migration of bone marrow mesenchymal stem cells was examined by PKH labeling. Caspase-3 activity was measured using spectrophotometry. The optimal neurological function recovery, lowest degree of ischemic cerebral damage, greatest number of bone marrow mesenchymal stem cells migrating to peri-ischemic area, and lowest caspase-3 activity in the ischemic cerebral tissue were observed in rats that underwent bone marrow mesenchymal stem cell transplantation at 12 hours after cerebral ischemia. These ifndings suggest that 12 hours after cerebral ischemia is the optimal time for tail vein injection of bone marrow mesenchymal stem cell transplantation against cerebral ischemia, and the strongest neuroprotective effect of this cell therapy appears at this time.

  17. Improving analytical methods for protein-protein interaction through implementation of chemically inducible dimerization.

    Science.gov (United States)

    Andersen, Tonni Grube; Nintemann, Sebastian J; Marek, Magdalena; Halkier, Barbara A; Schulz, Alexander; Burow, Meike

    2016-01-01

    When investigating interactions between two proteins with complementary reporter tags in yeast two-hybrid or split GFP assays, it remains troublesome to discriminate true- from false-negative results and challenging to compare the level of interaction across experiments. This leads to decreased sensitivity and renders analysis of weak or transient interactions difficult to perform. In this work, we describe the development of reporters that can be chemically induced to dimerize independently of the investigated interactions and thus alleviate these issues. We incorporated our reporters into the widely used split ubiquitin-, bimolecular fluorescence complementation (BiFC)- and Förster resonance energy transfer (FRET)- based methods and investigated different protein-protein interactions in yeast and plants. We demonstrate the functionality of this concept by the analysis of weakly interacting proteins from specialized metabolism in the model plant Arabidopsis thaliana. Our results illustrate that chemically induced dimerization can function as a built-in control for split-based systems that is easily implemented and allows for direct evaluation of functionality. PMID:27282591

  18. A crowdsourcing workflow for extracting chemical-induced disease relations from free text.

    Science.gov (United States)

    Li, Tong Shu; Bravo, Àlex; Furlong, Laura I; Good, Benjamin M; Su, Andrew I

    2016-01-01

    Relations between chemicals and diseases are one of the most queried biomedical interactions. Although expert manual curation is the standard method for extracting these relations from the literature, it is expensive and impractical to apply to large numbers of documents, and therefore alternative methods are required. We describe here a crowdsourcing workflow for extracting chemical-induced disease relations from free text as part of the BioCreative V Chemical Disease Relation challenge. Five non-expert workers on the CrowdFlower platform were shown each potential chemical-induced disease relation highlighted in the original source text and asked to make binary judgments about whether the text supported the relation. Worker responses were aggregated through voting, and relations receiving four or more votes were predicted as true. On the official evaluation dataset of 500 PubMed abstracts, the crowd attained a 0.505F-score (0.475 precision, 0.540 recall), with a maximum theoretical recall of 0.751 due to errors with named entity recognition. The total crowdsourcing cost was $1290.67 ($2.58 per abstract) and took a total of 7 h. A qualitative error analysis revealed that 46.66% of sampled errors were due to task limitations and gold standard errors, indicating that performance can still be improved. All code and results are publicly available athttps://github.com/SuLab/crowd_cid_relexDatabase URL:https://github.com/SuLab/crowd_cid_relex. PMID:27087308

  19. 骨髓间充质干细胞移植对慢性脑缺血大鼠海马区Cdc42表达及认知功能的影响%Transplantation of bone marrow mesenchymal stem cells increasing Cdc42 protein expression in the hippocampus and improving cognitive function of rats with chronic cerebral ischemia

    Institute of Scientific and Technical Information of China (English)

    于晓云; 张博爱; 李俊敏; 刘宇; 李晓晓; 崔璨; 崔红卫

    2013-01-01

    Objective To explore the effects of bone marrow mesenchymal stem cells transplantation on cognitive function and expression of cell division cycle 42 GTP-binding protein in hippocampus of chronic cerebral hypoperfusion rats. Methods 72 SD rats were randomly divided into sham, 2VO model (permanent ligation of bilateral common carotid artery) and experimental groups (2VO model + bone marrow mesenchymal stem cells transplantation). Rats in every group were subdivided into 8, 10 and 12 weeks subsets. Morris water maze was used to select rats of modeled successfully and space memory ability. Expression of Cdc42 protein in hippocampus was measured by Western blotting and immunohistochemistry. Results Expression of Cdc42 in hippocampus was reduced, escape latency was extended with time extending in experimental and model groups (P < 0. 05). Compared with model group, the expression of Cdc42 in the hippocampus tissue was increased and escape latency was shortened ( P < 0. 05 ) in experimental group. Conclusions Transplantation of bone marrow mesenchymal stem cells can improve cognitive function of chronic cerebral hypoperfusion rats, which maybe due to increased expression of Cdc42 protein.%目的 观察骨髓间充质干细胞移植后慢性脑缺血大鼠海马区表达Cdc42表达的变化,探讨其对慢性脑缺血大鼠认知功能的影响.方法 SD大鼠72只,随机分为假手术组、永久性双侧颈总动脉结扎(2VO)模型组、实验组(2VO模型+干细胞移植),每组分8、10、12w3个时间点,每个时间点8只,用Morris水迷宫筛选出造模成功的大鼠并检测大鼠的空间记忆能力,用Western印迹和免疫组化检测大鼠海马区Cdc42的表达.结果 实验组和模型组大鼠海马区Cdc42的表达随时间延长逐渐降低(P<0.05),其逃避潜伏期逐渐延长(P<0.05);较之2VO模型组,同一时间点实验组Cdc42表达明显增高(P<0.05),逃避潜伏期显著缩短(P<0.05).结论 骨髓间充质干细胞移植可显

  20. Influence of tail vein administration of bone marrow mesenchymal stem cells on expression of brain-derived neurotrophic factor and nerve growth factor after spinal cord injury in rats%骨髓间充质干细胞尾静脉移植脊髓损伤大鼠脑源性神经营养因子及神经生长因子的表达

    Institute of Scientific and Technical Information of China (English)

    董锋; 林建华; 吴朝阳

    2011-01-01

    BACKGROUND: Transplantation of bone marrow mesenchymal stem cells has an effect on the treatment of spinal cord injury, but the mechanism is not fully understood.OBJECTIVE: To explore the influence of intravenous administration of bone marrow mesenchymal stem cells on expression of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) after spinal cord injury in rats and to study possible mechanism of intravenous administration of bone marrow mesenchymal stem cells after spinal cord injury in adult rats.METHODS: Modified Allen method was applied to induce T 10 spinal cord injury to prepare rat models of paraplegia. There were 6 rats in sham operation group and 24 rats in injury group. Then, the injury group was randomly divided into transplantation group and control group. Rats in the sham operation and transplantation groups received tail vein transplantation of 1 mL bone marrow mesenchymal stem cells (1×106 cells), while those in the control group were treated with PBS.RESULTS AND CONCLUSION: After spinal cord injury in rats, the expression of BDNF and NGF was improved and advanced to increase via intravenous administration of bone marrow mesenchymal stem cells. Maybe it is one of the mechanisms of promoting restoration of injured spinal cord and improving motor function.%背景:骨髓间充质干细胞移植对脊髓损伤有治疗作用,但其机制尚不完全清楚.目的:应用免疫组织化学方法观察骨髓间充质干细胞静脉移植损伤脊髓局部脑源性神经营养因子及神经生长因子的表达,分析骨髓间充质干细胞移植治疗大鼠脊髓损伤的作用途径.方法:运用改良Allen 法制备T10 脊髓外伤性截瘫大鼠模型,假手术组6 只,脊髓损伤组24 只随机分为对照组和骨髓间充质干细胞移植组.骨髓间充质干细胞移植组、假手术组接受骨髓间充质干细胞单细胞悬液1 mL(1×106 cells)自大鼠尾静脉缓慢注射移植,对照组静脉注射PBS 1 mL.结果

  1. Bone marrow mesenchymal stem cells with Nogo-66 receptor gene silencing for repair of spinal cord injury

    OpenAIRE

    Li, Zhiyuan; Zhang, Zhanxiu; Zhao, Lili; LI Hui; Wang, Suxia; Shen, Yong

    2014-01-01

    We hypothesized that RNA interference to silence Nogo-66 receptor gene expression in bone marrow mesenchymal stem cells before transplantation might further improve neurological function in rats with spinal cord transection injury. After 2 weeks, the number of neurons and BrdU-positive cells in the Nogo-66 receptor gene silencing group was higher than in the bone marrow mesenchymal stem cell group, and significantly greater compared with the model group. After 4 weeks, behavioral performance ...

  2. MicroRNA-27b Enhances the Hepatic Regenerative Properties of Adipose-Derived Mesenchymal Stem Cells

    OpenAIRE

    Chen, Kuang-Den; Huang, Kuang-Tzu; Lin, Chih-Che; Weng, Wei-Teng; Hsu, Li-Wen; Goto, Shigeru; Nakano, Toshiaki; Lai, Chia-Yun; Kung, Chao-Pin; Chiu, King-Wah; Wang, Chih-Chi; Cheng, Yu-Fan; Ma, Yen-Ying; Chen, Chao-Long

    2016-01-01

    Adipose-derived mesenchymal stem cells (ASCs) are readily available multipotent mesenchymal progenitor cells and have become an attractive therapeutic tool for regenerative medicine. We herein investigated the mechanistic role of how miR-27b modulated regenerative capacities of ASCs. Intravenous administration of miR-27b-transfected ASCs (ASCs-miR-27b) was conducted after 70% partial hepatectomy (PH). After PH, rats injected with ASCs-miR-27b had decreased inflammatory cytokines and increased...

  3. Uses of mesenchymal stem cells

    OpenAIRE

    M. Delgado; González-Rey, Elena; Büscher, Dirk

    2008-01-01

    The invention relates to the use of mesenchymal stem cells (MSCs) for treating systemic infiammatory response syndrome (SIRS) in a subject. The invention provides compositions, uses and methods for the treatment of SIRS.

  4. Bone marrow mesenchymal stem cells repair spinal cord ischemia/reperfusion injur y by promoting axonal growth and anti-autophagy

    Institute of Scientific and Technical Information of China (English)

    Fei Yin; Chunyang Meng; Rifeng Lu; Lei Li; Ying Zhang; Hao Chen; Yonggang Qin; Li Guo

    2014-01-01

    Bone marrow mesenchymal stem cells can differentiate into neurons and astrocytes after trans-plantation in the spinal cord of rats with ischemia/reperfusion injury. Although bone marrow mesenchymal stem cells are known to protect against spinal cord ischemia/reperfusion injury through anti-apoptotic effects, the precise mechanisms remain unclear. In the present study, bone marrow mesenchymal stem cells were cultured and proliferated, then transplanted into rats with ischemia/reperfusion injury via retro-orbital injection. Immunohistochemistry and immunolfuorescence with subsequent quantiifcation revealed that the expression of the axonal regeneration marker, growth associated protein-43, and the neuronal marker, microtubule-as-sociated protein 2, significantly increased in rats with bone marrow mesenchymal stem cell transplantation compared with those in rats with spinal cord ischemia/reperfusion injury. Fur-thermore, the expression of the autophagy marker, microtubule-associated protein light chain 3B, and Beclin 1, was signiifcantly reduced in rats with the bone marrow mesenchymal stem cell transplantation compared with those in rats with spinal cord ischemia/reperfusion injury. Western blot analysis showed that the expression of growth associated protein-43 and neuro-iflament-H increased but light chain 3B and Beclin 1 decreased in rats with the bone marrow mesenchymal stem cell transplantation. Our results therefore suggest that bone marrow mes-enchymal stem cell transplantation promotes neurite growth and regeneration and prevents autophagy. These responses may likely be mechanisms underlying the protective effect of bone marrow mesenchymal stem cells against spinal cord ischemia/reperfusion injury.

  5. Transcriptome Sequencing of Chemically Induced Aquilaria sinensis to Identify Genes Related to Agarwood Formation

    Science.gov (United States)

    Ye, Wei; Wu, Hongqing; He, Xin; Wang, Lei; Zhang, Weimin; Li, Haohua; Fan, Yunfei; Tan, Guohui; Liu, Taomei; Gao, Xiaoxia

    2016-01-01

    Background Agarwood is a traditional Chinese medicine used as a clinical sedative, carminative, and antiemetic drug. Agarwood is formed in Aquilaria sinensis when A. sinensis trees are threatened by external physical, chemical injury or endophytic fungal irritation. However, the mechanism of agarwood formation via chemical induction remains unclear. In this study, we characterized the transcriptome of different parts of a chemically induced A. sinensis trunk sample with agarwood. The Illumina sequencing platform was used to identify the genes involved in agarwood formation. Methodology/Principal Findings A five-year-old Aquilaria sinensis treated by formic acid was selected. The white wood part (B1 sample), the transition part between agarwood and white wood (W2 sample), the agarwood part (J3 sample), and the rotten wood part (F5 sample) were collected for transcriptome sequencing. Accordingly, 54,685,634 clean reads, which were assembled into 83,467 unigenes, were obtained with a Q20 value of 97.5%. A total of 50,565 unigenes were annotated using the Nr, Nt, SWISS-PROT, KEGG, COG, and GO databases. In particular, 171,331,352 unigenes were annotated by various pathways, including the sesquiterpenoid (ko00909) and plant–pathogen interaction (ko03040) pathways. These pathways were related to sesquiterpenoid biosynthesis and defensive responses to chemical stimulation. Conclusions/Significance The transcriptome data of the different parts of the chemically induced A. sinensis trunk provide a rich source of materials for discovering and identifying the genes involved in sesquiterpenoid production and in defensive responses to chemical stimulation. This study is the first to use de novo sequencing and transcriptome assembly for different parts of chemically induced A. sinensis. Results demonstrate that the sesquiterpenoid biosynthesis pathway and WRKY transcription factor play important roles in agarwood formation via chemical induction. The comparative analysis of

  6. 骨髓间充质干细胞复合聚乳酸-羟基乙酸材料治疗大鼠神经损伤的实验研究%Bone Marrow Mesenchymal Stem Cells Combined with Polylactic Glycolic Acid Scaffold for Rat Neural Injury

    Institute of Scientific and Technical Information of China (English)

    符厚圣; 周兴; 桂有富; 郑煜; 潘建刚

    2011-01-01

    Objectives To study separately the effect of bone marrow mesenchymal stem cells (BMSCs) and differentiated neural - like cells combined with polylactic glycolic acid ( PLGA ) scaffolds, both composite which restore to SD rat nerve injury. Methods BMSCs were cultured by differential adherence,and composited PLGA,via scanning electron microscope to investigate the information of BMSCs'adhesion, proliferation and induction into neural - like cells in the scaffolds. Bold scaffold, BMSCs - PLGA complex, and neural - like cells - PLGA complex were separately transplanted into SD rat models, with the injury for right sciatic nerve, and obsvered SD rat models during 60 days after operation. Results BMSCs and neural - like cells can be exactly adhere to, proliferate and grow on the PLGA;BMSCs and neural -like cells combined with PLGA, which can restore to nerve injury. Conclusions BMSCs and differentiated neural - like cells combined with polylactic glycolic acid (PLGA) scaffolds,which can be used as a method of tissue engineering,to treat neurogenic bladder caused by spinal cord injury and advanced diabetes.%目的 研究骨髓间充质干细胞(Bone Marrow Mesenchymal Stem Cells,BMSCs)及其分化的神经样细胞分别与聚乳酸-羟基乙酸(polylactic glycolic acid,PLGA)支架材料复合修复大鼠神经损伤的效果.方法 将BMSCs复合PIGA培养,通过扫描电镜观察BMSCs在PLGA上的黏附、增殖并诱导分化成神经样细胞的情况;然后分别将单纯的PLGA、BMSCs-PLGA复合物以及神经样细胞-PLGA复合物分别移植入3组右侧坐骨神经损伤的SD大鼠模型中,术后观察三组动物60 d.结果 BMSCs及诱导分化生成的神经样细胞能在PLGA良好的黏附、增殖和生长;复合PLGA移植后能修复损伤的神经.结论 BMSCs及神经样细胞复合PLGA移植可作为组织工程治疗脊髓损伤及糖尿病晚期神经源性膀胱的一种方法.

  7. Myogenic Differentiation of Rat Bone Marrow Derived Mesenchymal Stem Cells Induced by 5-Azacytidine in Vitro%5-氮杂胞苷对大鼠骨髓间充质干细胞体外诱导分化的研究

    Institute of Scientific and Technical Information of China (English)

    孙思; 杜小文; 徐剑炜

    2012-01-01

    Objective:To explore the possibility of inducing myogenic differentiation of rat bone marrow derived mesenchymal stem cells (BMSCs) by 5-azacytidine in vitro. Methods: BMSCs were harvested from 1-week rats, and the surface antigens were identified by flow cytometer. The BMSCs were then treated with 5-azacytidine to achieve the myogenic differentiation. The expression of desmin and crsarcomeric aetin(α-SMA) of the induced BMSCs were detected by immunohistochemical staining. Results: BMSCs from rat long bones could be purified after 3 passages. CD29 and CD105 were positive, while CD34 and CD45 were negative. The immunohistochemical staining revealed the expression of desmin and crSMA in the inducted BMSCs. Conclusions: The myogenic differentiation of BMSCs could be induced by 5-azacytidine.%目的:探讨应用5-氮杂胞苷诱导大鼠骨髓间充质干细胞(bone marrow derived mesenchymal stem cells,BMSCs)向肌样细胞分化的可能性.方法:从1周龄SD大鼠骨髓提取分离BMSCs,在体外培养增殖,并采用流式细胞仪鉴定.以5-氮杂胞苷作用于BMSCs,诱导其向肌样细胞分化,继续培养3周后,采用苏木精-伊红、结蛋白、α-横纹肌动蛋白(α-sarcomeric actin,α-SMA)免疫组织化学染色观察BMSCs的分化情况.结果:从1周龄大鼠四肢长骨中提取骨髓,经贴壁法分离,传代后用流式细胞仪检测细胞表面抗体,结果显示CD29、CD105呈阳性,CD34、CD45呈阴性反应,P2、P3代BMSCs的比例明显提高.经诱导的细胞结蛋白、α-SMA染色呈强阳性.结论:在体外经5-氮杂胞苷诱导的BMSCs可向肌样细胞分化.

  8. Isolation, culture and identification of nucleus pulposus-derived mesenchymal stem cells from adult rats in vitro%大鼠椎间盘髓核来源间充质干细胞的体外分离及培养鉴定

    Institute of Scientific and Technical Information of China (English)

    芮云峰; 王善正; 谢鑫荟; 孙明辉; 林禹丞; 李刚; 王宸

    2013-01-01

    背景:目前椎间盘髓核组织的细胞组成和特性仍未阐明。  目的:旨在建立大鼠椎间盘髓核来源间充质干细胞的体外培养体系,并对其体外多项分化潜能进行鉴定。  方法:体外培养SD大鼠盘髓核来源间充质干细胞,取第3代细胞进行三系诱导分化,将成骨、成脂和成软骨分化作为实验组,基础细胞培养作为对照组。  结果与结论:低密度培养获得的髓核来源细胞早期可形成葵花样细胞集落,克隆样生长。第3代后,细胞形态趋向均一,呈成纤维细胞样生长。成骨诱导28 d,实验组茜素红染色阳性,且 RunX2、osteopontin 及osteocalcin表达较对照组显著增高(P OBJECTIVE:To establish the in vitro culture system of rat nucleus pulposus-derived mesenchymal stem cells and to identify their multi-lineage differentiation potential. METHODS:Mesenchymal stem cells from the nucleus pulposus tissues of Sprague-Dawley rats were cultured in vitro. Then, cells at passage 3 were induced to differentiate into osteoblasts, adipocytes and chondrocytes as experimental group. cells cultured with basic culture medium served as controls. RESULTS AND CONCLUSION:cells isolated from rat nucleus pulposus could form the sunflower-like colonies and exhibit clone-like growth when they cultured at a low density. cells at passage 3 became homogeneous and exhibited fibroblast-like morphology. After 28 days of osteogenic induction, arizarin red positive signals were detected in the experimental group. The mRNA expressions of RunX2, osteopontin and osteocalcin were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of adipogenic induction, oil red-O positive cells were detected in the experimental group. The mRNA expressions of C/EBPαand PPARγ2 were significantly increased in the experimental group, compared to the control group (P<0.05). After 21 days of chondrogenic

  9. 12 hours after cerebral ischemia is the optimal time for bone marrow mesenchymal stem cell transplantation

    OpenAIRE

    Seyed Mojtaba Hosseini; Mohammad Farahmandnia; Zahra Razi; Somayeh Delavarifar; Benafsheh Shakibajahromi

    2015-01-01

    Cell therapy using stem cell transplantation against cerebral ischemia has been reported. However, it remains controversial regarding the optimal time for cell transplantation and the transplantation route. Rat models of cerebral ischemia were established by occlusion of the middle cerebral artery. At 1, 12 hours, 1, 3, 5 and 7 days after cerebral ischemia, bone marrow mesenchymal stem cells were injected via the tail vein. At 28 days after cerebral ischemia, rat neurological function was eva...

  10. High Fidelity Tape Transfer Printing Based On Chemically Induced Adhesive Strength Modulation

    Science.gov (United States)

    Sim, Kyoseung; Chen, Song; Li, Yuhang; Kammoun, Mejdi; Peng, Yun; Xu, Minwei; Gao, Yang; Song, Jizhou; Zhang, Yingchun; Ardebili, Haleh; Yu, Cunjiang

    2015-11-01

    Transfer printing, a two-step process (i.e. picking up and printing) for heterogeneous integration, has been widely exploited for the fabrication of functional electronics system. To ensure a reliable process, strong adhesion for picking up and weak or no adhesion for printing are required. However, it is challenging to meet the requirements of switchable stamp adhesion. Here we introduce a simple, high fidelity process, namely tape transfer printing(TTP), enabled by chemically induced dramatic modulation in tape adhesive strength. We describe the working mechanism of the adhesion modulation that governs this process and demonstrate the method by high fidelity tape transfer printing several types of materials and devices, including Si pellets arrays, photodetector arrays, and electromyography (EMG) sensors, from their preparation substrates to various alien substrates. High fidelity tape transfer printing of components onto curvilinear surfaces is also illustrated.

  11. Choice of mouse strain influences the outcome in a mouse model of chemical-induced asthma.

    Directory of Open Access Journals (Sweden)

    Vanessa De Vooght

    Full Text Available BACKGROUND: The development of occupational asthma is the result of interactions between environmental factors and individual susceptibility. We assessed how our model of chemical-induced asthma is influenced by using different mouse strains. METHODOLOGY/PRINCIPAL FINDINGS: On days 1 and 8, male mice of 7 different strains (BALB/c, BP/2, A/J, C57Bl/6, DBA/2, CBA and AKR were dermally treated with toluene-2,4-diisocyanate (TDI (0.3% or vehicle (acetone/olive oil, AOO, 2:3 on each ear (20 microl. On day 15, they received an oropharyngeal instillation of TDI (0.01% or AOO (1:4. Airway reactivity to methacholine, total and differential cell counts in bronchoalveolar lavage (BAL and total serum IgE and IgG(2a levels were measured. Lymphocyte subpopulations in auricular lymph nodes and in vitro release of cytokines by ConA stimulated lymphocytes were assessed. In TDI-sensitized and challenged mice, airway hyper-reactivity was only observed in BALB/c, BP/2, A/J and AKR mice; airway inflammation was most pronounced in BALB/c mice; numbers of T-helper (CD4(+, T-activated (CD4(+CD25(+, T-cytotoxic (CD8(+ and B- lymphocytes (CD19(+ were increased in the auricular lymph nodes of BALB/c, BP/2, A/J and CBA mice; elevated concentrations of IL-4, IL-10, IL-13 and IFN-gamma were detected in supernatant of lymphocytes from BALB/c, BP/2, A/J, C57Bl/6 and CBA mice cultured with concanavaline A, along with an increase in total serum IgE. CONCLUSION: The used mouse strain has considerable and variable impacts on different aspects of the asthma phenotype. The human phenotypical characteristics of chemically-induced occupational asthma were best reproduced in Th2-biased mice and in particular in BALB/c mice.

  12. Topical application of silymarin reduces chemical-induced irritant contact dermatitis in BALB/c mice.

    Science.gov (United States)

    Han, Mi Hwa; Yoon, Won Kee; Lee, Hyunju; Han, Sang-Bae; Lee, Kiho; Park, Song-Kyu; Yang, Kyu-Hwan; Kim, Hwan Mook; Kang, Jong Soon

    2007-12-15

    Irritant contact dermatitis (ICD) is a non-allergic local inflammatory reaction of a skin and one of the most frequent occupational health problems. Silymarin has been clinically used in Europe for a long time to treat liver diseases and also known to have anti-cancer and anti-inflammatory activities. In the present study, we report that topical application of silymarin reduces chemical-induced ICD. Topical application of 2,4-dinitrochlorobenzene (DNCB) induced an ear swelling in BALB/c mice and silymarin suppressed DNCB-induced increase in ear thickness. Prophylactic and therapeutic application of silymarin showed similar effect on DNCB-induced increase in ear thickness and skin water content. In addition, phobor ester- or croton oil-induced increase in ear thickness was also inhibited by silymarin treatment. Silymarin also blocked neutrophil accumulation into the ear induced by these irritants. Further study demonstrated that DNCB-induced tumor necrosis factor-alpha (TNF-alpha) expression in mouse ear was suppressed by silymarin. DNCB-induced expression of KC, one of the main attractors of neutrophil in mice, and adhesion molecules, including intercellular adhesion molecule-1 (ICAM-1) and E-selectin in mouse ear were also inhibited by silymarin. Moreover, TNF-alpha-induced expression of cytokines, such as TNF-alpha and IL-1beta, and a chemokine, IL-8, were suppressed by silymarin treatment in human keratinocyte cell line, HaCaT. Silymarin also blocked TNF-alpha- and DNCB-induced NF-kappaB activation in HaCaT. Collectively, these results demonstrate that topically applied silymarin inhibits chemical-induced ICD in mice and this might be mediated, at least in part, by blocking NF-kappaB activation and consequently inhibiting the expression of cytokines and adhesion molecules. PMID:17996674

  13. Histopathological image analysis of chemical-induced hepatocellular hypertrophy in mice.

    Science.gov (United States)

    Asaoka, Yoshiji; Togashi, Yuko; Mutsuga, Mayu; Imura, Naoko; Miyoshi, Tomoya; Miyamoto, Yohei

    2016-04-01

    Chemical-induced hepatocellular hypertrophy is frequently observed in rodents, and is mostly caused by the induction of phase I and phase II drug metabolic enzymes and peroxisomal lipid metabolic enzymes. Liver weight is a sensitive and commonly used marker for detecting hepatocellular hypertrophy, but is also increased by a number of other factors. Histopathological observations subjectively detect changes such as hepatocellular hypertrophy based on the size of a hepatocyte. Therefore, quantitative microscopic observations are required to evaluate histopathological alterations objectively. In the present study, we developed a novel quantitative method for an image analysis of hepatocellular hypertrophy using liver sections stained with hematoxylin and eosin, and demonstrated its usefulness for evaluating hepatocellular hypertrophy induced by phenobarbital (a phase I and phase II enzyme inducer) and clofibrate (a peroxisomal enzyme inducer) in mice. The algorithm of this imaging analysis was designed to recognize an individual hepatocyte through a combination of pixel-based and object-based analyses. Hepatocellular nuclei and the surrounding non-hepatocellular cells were recognized by the pixel-based analysis, while the areas of the recognized hepatocellular nuclei were then expanded until they ran against their expanding neighboring hepatocytes and surrounding non-hepatocellular cells by the object-based analysis. The expanded area of each hepatocellular nucleus was regarded as the size of an individual hepatocyte. The results of this imaging analysis showed that changes in the sizes of hepatocytes corresponded with histopathological observations in phenobarbital and clofibrate-treated mice, and revealed a correlation between hepatocyte size and liver weight. In conclusion, our novel image analysis method is very useful for quantitative evaluations of chemical-induced hepatocellular hypertrophy. PMID:26776450

  14. An experimental study on repairing sciatic nerve defects of rats by human umbilical cord derived mesenchymal stem cells and acellular nerve basal lamina tube%人脐带间充质干细胞复合去细胞神经基膜管修复大鼠坐骨神经缺损

    Institute of Scientific and Technical Information of China (English)

    吴学建; 何江涛; 孙士强

    2010-01-01

    Objective To explore the feasibility of using human umbilical cord derived mesenchymal stem cells as seed cells to repair sciatic nerve defects of rats by tissue engineering methods. Methods Mesenchymal stem cells from human umbilical cord were cultured and induced into neuron-liked cells,which were co-cultured with acellular basal lamina tube to construct tissue engineering nerve;models of sciatic nerve defects 10 mm in length were set up with thirty healthy adult SD rats and were divided randomly into 3 groups:tissue engineering nerve group (group A, compound of human umbilical cord derived mesenchymal stem cells and acellular basal lamina tube), pure acellular basal lamina tube group (group B), and autogenous nerve bridging group (group C). Evaluation of electrophysiological and histological results was carried out 10 weeks after operation. Results The engineering nerve group had good result in nerve regeneration which was close to the effect of autogenous nerve transfer group (group A), and much better than the effect of pure acellular basal lamina tube group. Conclusion Engineering nerves from human umbilical cord derived mesenchymal stem cells can effectively repair 10 mm defects of sciatic nerve.%目的 探讨诱导后的脐带间充质干细胞作为组织工程种子细胞修复大鼠坐骨神经缺损的可行性.方法 从正常足月新生儿脐带中分离培养间充质干细胞并诱导分化为神经样细胞,与去细胞神经基膜管共培养以构建组织工程神经;用30只健康成年雄性SD大鼠建立坐骨神经缺损(10 mm)的动物模型并随机分成3组:A组为脐带间充质干细胞复合去细胞神经基膜管组,B组为单纯去细胞神经基膜管组,C组为自体神经桥接组.术后10周通过神经电生理检测、组织学观察等评测效果.结果 在局部观察和肌肉测量、神经电生理检测、组织学观察等方面,脐带间充质干细胞复合去细胞神经基膜管组(A组)神经再生及肢体功

  15. 骨髓间充质干细胞对脓毒症急性肺损伤大鼠肺泡巨噬细胞NF-κB的调控%Bone marrow mesenchymal stem cells regulate nuclear factor kappaB expression in alveolar macrophages of acute lung injury rats with sepsis

    Institute of Scientific and Technical Information of China (English)

    张继峰; 张紫琦; 雒晓甜; 侯林义; 姜琴; 吕洁萍; 张文凯

    2015-01-01

    BACKGROUND:Bone marrow mesenchymal stem cels have a therapeutic effect on acute lung injury, but the mechanism is unclear. If the mechanism is understood, the majority of patients with acute lung injury can obtain a benefit. OBJECTIVE:To explore the possible mechanism underlying bone marrow mesenchymal stem cels in the treatment of acute lung injury with sepsis in rats. METHODS: (1) Thirty-six adult Wistar rats were randomly divided into three groups, sham operation group (sham group), sepsis group and bone marrow mesenchymal stem cels group (cel treatment group). In the sepsis and cel treatment groups, animal models of sepsis with acute lung injury were established by cecal ligation and puncture, while in the sham group, the cecum was not ligated and punctured. Then, 1 mL normal saline was injected via the femoral vein in the sepsis and sham groups, and 1 mL bone marrow mesenchymal stem cel suspension (1×109/L) was injected into the cel treatment group. After 6 hours, interleukin 10 and macrophage inflammatory protein-2 levels in serum were measured in the three groups. Lung tissues were taken for pathological observation using hematoxylin-eosin staining. (2) Rat alveolar macrophages were obtained by bronchoalveolar lavage, seeded into 24-wel culture plates, and divided into three groups: control group (group A), sepsis model group (group B) and intervention group of bone marrow mesenchymal stem cels (group C). Normal saline, septic plasma, and co-intervention of septic plasma and mesenchymal stem cels were used in the groups A, B, C, respectively. Then, cels in the three groups were cultured in a 5% CO2 incubator at 37℃ for 1 hour. After that, alveolar macrophages were taken to detect whether nuclear factor-κB (P65) protein entered into the nucleus using laser scanning confocal microscopy. RESULTS AND CONCLUSION: (1) The results of animal experiments showed that compared with the sham group, the macrophage inflammatory protein-2 levels in the sepsis group and

  16. Real-time-guided bone regeneration around standardized critical size calvarial defects using bone marrow-derived mesenchymal stem cells and collagen membrane with and without using tricalcium phosphate: an in vivo micro-computed tomographic and histologic experiment in rats.

    Science.gov (United States)

    Al-Hezaimi, Khalid; Ramalingam, Sundar; Al-Askar, Mansour; ArRejaie, Aws S; Nooh, Nasser; Jawad, Fawad; Aldahmash, Abdullah; Atteya, Muhammad; Wang, Cun-Yu

    2016-01-01

    The aim of the present real time in vivo micro-computed tomography (µCT) and histologic experiment was to assess the efficacy of guided bone regeneration (GBR) around standardized calvarial critical size defects (CSD) using bone marrow-derived mesenchymal stem cells (BMSCs), and collagen membrane (CM) with and without tricalcium phosphate (TCP) graft material. In the calvaria of nine female Sprague-Dawley rats, full-thickness CSD (diameter 4.6 mm) were created under general anesthesia. Treatment-wise, rats were divided into three groups. In group 1, CSD was covered with a resorbable CM; in group 2, BMSCs were filled in CSD and covered with CM; and in group 3, TCP soaked in BMSCs was placed in CSD and covered with CM. All defects were closed using resorbable sutures. Bone volume and bone mineral density of newly formed bone (NFB) and remaining TCP particles and rate of new bone formation was determined at baseline, 2, 4, 6, and 10 weeks using in vivo µCT. At the 10th week, the rats were killed and calvarial segments were assessed histologically. The results showed that the hardness of NFB was similar to that of the native bone in groups 1 and 2 as compared to the NFB in group 3. Likewise, values for the modulus of elasticity were also significantly higher in group 3 compared to groups 1 and 2. This suggests that TCP when used in combination with BMSCs and without CM was unable to form bone of significant strength that could possibly provide mechanical "lock" between the natural bone and NFB. The use of BMSCs as adjuncts to conventional GBR initiated new bone formation as early as 2 weeks of treatment compared to when GBR is attempted without adjunct BMSC therapy. PMID:27025260

  17. Mesenchymal stem cells protect from hypoxia-induced alveolar epithelial-mesenchymal transition.

    Science.gov (United States)

    Uzunhan, Yurdagül; Bernard, Olivier; Marchant, Dominique; Dard, Nicolas; Vanneaux, Valérie; Larghero, Jérôme; Gille, Thomas; Clerici, Christine; Valeyre, Dominique; Nunes, Hilario; Boncoeur, Emilie; Planès, Carole

    2016-03-01

    Administration of bone marrow-derived human mesenchymal stem cells (hMSC) reduces lung inflammation, fibrosis, and mortality in animal models of lung injury, by a mechanism not completely understood. We investigated whether hMSC would prevent epithelial-mesenchymal transition (EMT) induced by hypoxia in primary rat alveolar epithelial cell (AEC). In AEC cultured on semipermeable filters, prolonged hypoxic exposure (1.5% O2 for up to 12 days) induced phenotypic changes consistent with EMT, i.e., a change in cell morphology, a decrease in transepithelial resistance (Rte) and in the expression of epithelial markers [zonula occludens-1 (ZO-1), E-cadherin, AQP-5, TTF-1], together with an increase in mesenchymal markers [vimentin, α-smooth muscle actin (α-SMA)]. Expression of transcription factors driving EMT such as SNAIL1, ZEB1, and TWIST1 increased after 2, 24, and 48 h of hypoxia, respectively. Hypoxia also induced TGF-β1 mRNA expression and the secretion of active TGF-β1 in apical medium, and the expression of connective tissue growth factor (CTGF), two inducers of EMT. Coculture of AEC with hMSC partially prevented the decrease in Rte and in ZO-1, E-cadherin, and TTF-1 expression, and the increase in vimentin expression induced by hypoxia. It also abolished the increase in TGF-β1 expression and in TGF-β1-induced genes ZEB1, TWIST1, and CTGF. Finally, incubation with human recombinant KGF at a concentration similar to what was measured in hMSC-conditioned media restored the expression of TTF-1 and prevented the increase in TWIST1, TGF-β1, and CTGF in hypoxic AEC. Our results indicate that hMSC prevent hypoxia-induced alveolar EMT through the paracrine modulation of EMT signaling pathways and suggest that this effect is partly mediated by KGF. PMID:26702148

  18. visual bone marrow mesenchymal stem cell transplantation in the repair of spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    Rui-ping Zhang; Cheng Xu; Yin Liu; Jian-ding Li; Jun Xie

    2015-01-01

    An important factor in improving functional recovery from spinal cord injury using stem cells is maximizing the number of transplanted cells at the lesion site. Here, we established a contusion model of spinal cord injury by dropping a weight onto the spinal cord at T7–8. Superparamagnet-ic iron oxide-labeled bone marrow mesenchymal stem cells were transplanted into the injured spinal cordvia the subarachnoid space. An outer magnetic ifeld was used to successfully guide the labeled cells to the lesion site. Prussian blue staining showed that more bone marrow mesen-chymal stem cells reached the lesion site in these rats than in those without magnetic guidance or superparamagnetic iron oxide labeling, and immunolfuorescence revealed a greater number of complete axons at the lesion site. Moreover, the Basso, Beattie and Bresnahan (BBB) locomotor rating scale scores were the highest in rats with superparamagnetic labeling and magnetic guid-ance. Our data conifrm that superparamagnetic iron oxide nanoparticles effectively label bone marrow mesenchymal stem cells and impart sufficient magnetism to respond to the external magnetic ifeld guides. More importantly, superparamagnetic iron oxide-labeled bone marrow mesenchymal stem cells can be dynamically and non-invasively trackedin vivo using magnetic resonance imaging. Superparamagnetic iron oxide labeling of bone marrow mesenchymal stem cells coupled with magnetic guidance offers a promising avenue for the clinical treatment of spinal cord injury.

  19. 与盆底韧带成纤维细胞间接共培养对大鼠骨髓间充质干细胞弹性蛋白、LOX及Fibulin -5mRNA表达的影响%Effect of indirect coculture of fibroblasts in pelvic ligament and bone marrow mesenchymal stem cells on the expressions of elastin, LOX, and Fibulin -5 mRNA in bone marrow mesenchymal stem cells of rats

    Institute of Scientific and Technical Information of China (English)

    梁琳琳; 闫建国; 任琛琛; 赵冰; 张翠莲

    2012-01-01

    Objective: To study Ihe effect of indirect coculture of fibroblasts in pelvic ligament on differentiation of bone marrow mesenchymal stem cells (BMSCs) of rats, explore the establishment of differentiation microenvironment of inducing BMSCs to fibroblasts in pelvic ligament, detect the expressions of elaslin, LOX, and Fibulin -5. Methods: Seven - day SD rats were selected, density gradient centrifugation was used to separate and purify BMSCs of rats and determine BMSCs; the pelvic ligament tissue of rats were obtained, separated and passaged, then fibroblasls were obtained; the purified fourth generation fibroblasts of pelvic ligament and the fourth generation BMSCs after purification and culture were cocultured indirectly; real - time quantitative RT - PCR was used to detect the expressions of elastin, LOX, and Fibulin -5 mRNA after coculture for different days. Results:Compared with negative control group after coculture for the same days, the expression levels of elastin, LOX, and Fibulin -S mRNA increased after coculture for three days, but there was no significant difference (P > 0.05). At six and twelve days after coculture, the expression levels of elastin, LOX, and Fibulin - 5 mRNA increased significantly (P 0.05). Compared with three - day coculture group, the expression levels of elastin, LOX, and Fibulin - 5 mRNA in six - day coculture group increased, but there was no significant difference (P > 0.05), but the expression levels of elastin, LOX, and Fibulin - 5 mRNA in twelve - day coculture group increased significantly ( P < 0.05). Conclusion: Coculture of fibroblasts in pelvic ligament and BMSCs can promote the expression levels of elastin, LOX, and Fibulin -5 mRNA, which provide a new thought for exploring the differentiation conditions of BMSCs in vitro as seed cells in ligament tissue project.%目的:研究与盆底韧带成纤维细胞间接共培养对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)分化的影响,探索

  20. 转染血管内皮生长因子基因的骨髓间充质干细胞修复糖尿病大鼠足背创面%Transplantation of bone marrow mesenchymal stem cells transfected by vascular endothelial growth factor gene promotes foot wound healing in diabetic rats

    Institute of Scientific and Technical Information of China (English)

    蔡黔; 万江波; 梁文佳; 刘毅

    2014-01-01

    transplantation of bone marrow mesenchymal stem cells transfected by human vascular endothelial growth factor 165 (hVEGF165) gene on foot wound healing in diabetic rats. METHODS:Recombinant adenovirus was established in vitro which expressed hVEGF165 gene and transfected into the third generation of bone marrow mesenchymal stem cells. Total y 120 male Wistar rats were divided into five groups:group A (non-diabetic controls), group B (diabetic controls), group C (Ad-hVEGF165 therapy), group D (stem celltherapy) and group E (transplantation of bone marrow mesenchymal stem cells transfected by Ad-hVEGF165 gene). Rats in the latter four groups were intraperitoneal y injected with streptozotocin to induce diabetic models. In al rats, a 3 mm×7 mm rectangular ful-thickness skin sample was cut from the instep of the hind foot to make a model of foot wound. The rats were subcutaneously injected at equidistant six points 5 mm distal to the wound edge on the dorsum of the foot:50μL PBS per point for group A, 50μL adenovirus suspension (1×1013 pfu/L) per point for group C, 50μL stem cellsuspension (1×1010/L) per point for group D, and 50μL adenovirus suspension+50μL stem cellsuspension per point for group E. RESULTS AND CONCLUSION:After injection, the rate of wound healing, the expression of VEGF and the qualities of capil aries in group E were higher when compared with groups B, C, D (P<0.05), but were lower than those in group A (P<0.05). Transplantation of bone marrow mesenchymal stem cells transfected by hVEGF165 gene can promote foot wound healing, angiogenesis and expression of VEGF in diabetic rats.

  1. Imaging the impact of chemically inducible proteins on cellular dynamics in vivo.

    Directory of Open Access Journals (Sweden)

    Hon S Leong

    Full Text Available The analysis of dynamic events in the tumor microenvironment during cancer progression is limited by the complexity of current in vivo imaging models. This is coupled with an inability to rapidly modulate and visualize protein activity in real time and to understand the consequence of these perturbations in vivo. We developed an intravital imaging approach that allows the rapid induction and subsequent depletion of target protein levels within human cancer xenografts while assessing the impact on cell behavior and morphology in real time. A conditionally stabilized fluorescent E-cadherin chimera was expressed in metastatic breast cancer cells, and the impact of E-cadherin induction and depletion was visualized using real-time confocal microscopy in a xenograft avian embryo model. We demonstrate the assessment of protein localization, cell morphology and migration in cells undergoing epithelial-mesenchymal and mesenchymal-epithelial transitions in breast tumors. This technique allows for precise control over protein activity in vivo while permitting the temporal analysis of dynamic biophysical parameters.

  2. Rat bone marrow mesenchymal stem cells in vitro cultivation and establishing pEGFP/Ang-1 transfection method%大鼠骨髓间充质干细胞体外培养与pEGFP/Ang-1转染方法的建立

    Institute of Scientific and Technical Information of China (English)

    李福智; 侯阳; 李晓明; 房艳; 臧东钰

    2014-01-01

    Objective To train the successful bone marrow mesenchymal stem cells ( BMSCs ) and phenotypic characterization , and investigate the beneficial effects of recombinant plasmid ( pEGFP/Ang-1) transfected rats BMSCs .Methods Using adherence screening and density gradient centrifugation methods , purified rats BMSCs were obtained .After transfection with pEGFP/Ang-1, the BMSCs were screened with G418, and the positive clones were sequentially confirmed by expression of enhanced green fluorescent protein (GFP) with fluorescence microscope, and expression of angiogenin-1(Ang-1) was tested by Western blot.Results Using adherence screening and density gradient centrifugation methods , purified rats BMSCs were successfully obtained .The successfully stable transfection of pEGFP-N1-Ang-1 in BMSCs had be stably expressed Ang-1 3 days,later started to diminish.Conclusions The successful use of adherent culture method and density gra-dient centrifugation of rat BMSCs and the recombinant plasmid pEGFP /Ang-1 being transfected into the rat BMSCs could effectively express .%目的:探讨培养大鼠骨髓间充质干细胞(BMSCs)细胞株、表型鉴定及利用重组质粒(pEGFP/Ang-1)转染大鼠 BMSCs细胞株的实验方法。方法采用贴壁筛选法和密度梯度离心法获取BMSCs,纯化大鼠BMSCs后,以重组质粒pEGFP/Ang-1转染,G 418筛挑阳性克隆,倒置荧光显微镜下观察增强型绿色荧光蛋白的表达,检测Ang-1表达量的时间变化。结果采用贴壁筛选法和密度梯度离心法成功获取BMSCs细胞株,成功将血管生成素-1重组质粒(pEGFP/Ang-1)转染大鼠BMSCs,3 d可稳定表达Ang-1,7 d以后开始减弱。结论成功运用贴壁筛选法和密度梯度离心法分离培养的大鼠BMSCs,并重组质粒pEGFP/Ang-1转染大鼠BMSCs可有效表达。

  3. H-2 restriction of the T cell response to chemically induced tumors: evidence from F1 → parent chimeras

    International Nuclear Information System (INIS)

    It has been well established that T cells that react to tumor antigen on virus-induced tumors must share H-2D or H-2K specificities with the tumor. It has been impossible to perform similar studies with chemically induced tumors because each chemically induced tumor expresses a unique tumor antigen that cannot be studied in association with other H-2 types. This study provies evidence that H-2 recognition is also necessary for recognition of chemically induced tumors. We have found that F1 → parent chimeras preferentially recognize chemically induced tumors of parental H-2 type. C3H/HeJ and C57BL/6 mice were lethally irradiated and restored with (C3H x C57BL/6) F1 hybrid bone marrow. The F1 → C3H chimera but not the F1 → C57BL/6 chimera was able to respond to a C3H fibrosarcoma in mixed lymphocyte-tumor cell culture and also to neutralize the tumor in an in vivo tumor neutralization assay. On the other hand, the F1 → C57BL/6 chimera but not the F1 → C3H chimera was able to kill the C57BL/6 lymphoma EL4 in an in vitro cytotoxicity assay. Both chimeras were tolerant to C3H and C57BL/6 alloantigens but could respond normally to Con A and to BALB/c spleen cells in mixed lymphocyte cultures and cytotoxicity assay

  4. 甲钴胺体外诱导大鼠骨髓间充质干细胞向神经元样细胞分化☆%Methylcobalamin induces differentiation of rat bone mesenchymal stem cells into neuron-like cells in vitro

    Institute of Scientific and Technical Information of China (English)

    羊明智; 彭立军; 胡文凯

    2013-01-01

      背景:目前骨髓间充质干细胞移植到脊髓损伤动物体内后对脊髓损伤的恢复效果非常有限。甲钴胺是治疗神经系统疾病及损伤的常见药物,其对骨髓间充质干细胞的影响尚不清楚。目的:探讨甲钴胺体外定向诱导骨髓间充质干细胞向神经元样细胞分化的可行性,观察分化后的细胞增殖和生长情况。方法:取大鼠胫腓骨骨髓,采用密度梯度离心贴壁细胞培养法分离、培养骨髓间充质干细胞,取第四五代骨髓间充质干细胞,分别以25,50,100 mg/L的甲钴胺进行诱导分化24,48和72 h。倒置相差显微镜下连续观察细胞形态学变化,MTT法检测细胞活性,RT-PCR和Western blot法检测特异性标志物Nestin和NSE的表达。结果与结论:甲钴胺诱导骨髓间充质干细胞后大部分细胞变成神经元样细胞。与对照组比较,甲钴胺诱导后细胞活性无明显变化。不同剂量甲钴胺诱导48 h后,Nestin和NSE在mRNA和蛋白水平表达均上调,其中100 mg/L组表达上调最明显;100 mg/L 甲钴胺诱导24,48和72 h后,Nestin和NSE在mRNA和蛋白水平表达均上调,其中72 h表达上调最明显。说明甲钴胺可定向诱导骨髓间充质干细胞向神经元样细胞分化,100 mg/L为甲钴胺的较佳诱导浓度。%BACKGROUND:Currently, transplantation of bone marrow mesenchymal stem cel s into the spinal cord is very limited to the recovery of animals fol owing spinal cord injury. Methylcobalamin is a common drug for the treatment of neurological diseases and injuries, but its effects on bone marrow mesenchymal stem cel s are unclear. OBJECTIVE:To study the feasibility of bone marrow mesenchymal stem cel s differentiating into neuron-like cel s induced by methylcobalamin in vitro and to observe the cel viability and proliferation of differentiated cel s. Methods:Rat bone marrow mesenchymal stem cel s were isolated, cultured and purified by

  5. Combining machine learning, crowdsourcing and expert knowledge to detect chemical-induced diseases in text

    Science.gov (United States)

    Bravo, Àlex; Li, Tong Shu; Su, Andrew I.; Good, Benjamin M.; Furlong, Laura I.

    2016-01-01

    Drug toxicity is a major concern for both regulatory agencies and the pharmaceutical industry. In this context, text-mining methods for the identification of drug side effects from free text are key for the development of up-to-date knowledge sources on drug adverse reactions. We present a new system for identification of drug side effects from the literature that combines three approaches: machine learning, rule- and knowledge-based approaches. This system has been developed to address the Task 3.B of Biocreative V challenge (BC5) dealing with Chemical-induced Disease (CID) relations. The first two approaches focus on identifying relations at the sentence-level, while the knowledge-based approach is applied both at sentence and abstract levels. The machine learning method is based on the BeFree system using two corpora as training data: the annotated data provided by the CID task organizers and a new CID corpus developed by crowdsourcing. Different combinations of results from the three strategies were selected for each run of the challenge. In the final evaluation setting, the system achieved the highest Recall of the challenge (63%). By performing an error analysis, we identified the main causes of misclassifications and areas for improving of our system, and highlighted the need of consistent gold standard data sets for advancing the state of the art in text mining of drug side effects. Database URL: https://zenodo.org/record/29887?ln¼en#.VsL3yDLWR_V PMID:27307137

  6. Combining machine learning, crowdsourcing and expert knowledge to detect chemical-induced diseases in text.

    Science.gov (United States)

    Bravo, Àlex; Li, Tong Shu; Su, Andrew I; Good, Benjamin M; Furlong, Laura I

    2016-01-01

    Drug toxicity is a major concern for both regulatory agencies and the pharmaceutical industry. In this context, text-mining methods for the identification of drug side effects from free text are key for the development of up-to-date knowledge sources on drug adverse reactions. We present a new system for identification of drug side effects from the literature that combines three approaches: machine learning, rule- and knowledge-based approaches. This system has been developed to address the Task 3.B of Biocreative V challenge (BC5) dealing with Chemical-induced Disease (CID) relations. The first two approaches focus on identifying relations at the sentence-level, while the knowledge-based approach is applied both at sentence and abstract levels. The machine learning method is based on the BeFree system using two corpora as training data: the annotated data provided by the CID task organizers and a new CID corpus developed by crowdsourcing. Different combinations of results from the three strategies were selected for each run of the challenge. In the final evaluation setting, the system achieved the highest Recall of the challenge (63%). By performing an error analysis, we identified the main causes of misclassifications and areas for improving of our system, and highlighted the need of consistent gold standard data sets for advancing the state of the art in text mining of drug side effects.Database URL: https://zenodo.org/record/29887?ln¼en#.VsL3yDLWR_V. PMID:27307137

  7. Heuristic model of chemically induced electron spin polarization in two dimensions

    International Nuclear Information System (INIS)

    Graphical abstract: Like its three-dimensional predecessor, and unlike previous 2D models, this model handles all singlet-triplet mixing rates (Q) and, as illustrated below, gives the time evolution of the polarization. - Abstract: A heuristic model of chemically induced electron spin polarization (CIDEP) that breaks the polarization mechanism into its component steps, with each step governed by an appropriate solution of the diffusion equation, is extended from a three to a two-dimensional system. The required solution of the 2D diffusion equation is provided by a relatively simple analytic approximation to the usual infinite series solution. The model yields the polarization and its time development for weak to strong singlet-triplet mixing in the radical pairs, whereas previous models are limited to very weak or very strong mixing. Its results agree with a variational solution of an integral equation of Monchick and are encouraging for observation of CIDEP in dimensionally restricted systems. The method also may be applicable to other diffusion-controlled, spin-dependent chemistry in spatially restricted environments.

  8. Modulatory influence of Phyllanthus niruri on oxidative stress, antioxidant defense and chemically induced skin tumors.

    Science.gov (United States)

    Sharma, Priyanka; Parmar, Jyoti; Verma, Preeti; Goyal, Pradeep Kumar

    2011-01-01

    The present study evaluates the modulatory potential of Phyllanthus niruri on chemically induced skin carcinogenesis, and its influence on oxidative stress and the antioxidant defense system. Oral administration of P. niruri extract (PNE), during peri- (Gr. III), post- (Gr. IV), or peri- and post- (Gr. V) initiational stages of 7,12-dimethylbenz(a) anthracene (DMBA)-croton oil–induced papillomagenesis considerably reduced tumor burden to 4.20, 4.00, and 3.33(positive control value 6.20); cumulative number of papillomas to 21, 16, and 10, respectively, (positive control value 62); and incidence of mice bearing papillomas to 50, 40, and 30%, respectively (positive control value 100%), but significantly increased the average latent period to 10.14, 10.62, and 11.60, and inhibition of tumor multiplicity to 66, 74,and 83%, respectively. Enzyme analysis of skin and liver showed a significant (p ≤ 0.05, ≤ 0.01, ≤ 0.001) elevation in antioxidant parameters such as superoxide dismutase, catalase, glutathione, and vitamin C in PNE-treated groups (Gr. III–V) when compared with the carcinogen-treated control (Gr. II). The elevated level of lipid peroxidation in the carcinogen-treated positive control group was significantly (p ≤ 0.05, ≤ 0.01, ≤ 0.001) inhibited by PNE administration. These results indicate that P. niruri extract has potentiality to reduce skin papillomas by enhancing antioxidant defense system. PMID:21609315

  9. Insights from advances in research of chemically induced experimental models of human inflammatory bowel disease

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Inflammatory bowel disease (IBD), the most important being Crohn's disease and ulcerative colitis, results from chronic dysregulation of the mucosal immune system in the gastrointestinal tract. Although the pathogenesis of IBD remains unclear, it is widely accepted that genetic, environmental, and immunological factors are involved. Recent studies suggest that intestinal epithelial defenses are important to prevent inflammation by protecting against microbial pathogens and oxidative stresses. To investigate the etiology of IBD, animal models of experimental colitis have been developed and are frequently used to evaluate new anti-inflammatory treatments for IBD. Several models of experimental colitis that demonstrate various pathophysiological aspects of the human disease have been described. In this manuscript, we review the characteristic features of IBD through a discussion of the various chemically induced experimental models of colitis (e.g. dextran sodium sulfate-, 2,4,6-trinitrobenzene sulfonic acid-, oxazolone-, acetic acid-, and indomethacin-induced models). We also summarize some regulatory and pathogenic factors demonstrated by these models that can, hopefully, be exploited to develop future therapeutic strategies against IBD.

  10. The relationship between chemically-induced meiotic delay and aneuploidy in mouse oocytes and zygotes

    Energy Technology Data Exchange (ETDEWEB)

    Mailhes, J.B.; Marchetti, F. [Louisiana State Univ. Medical Center, Shreveport, LA (United States)

    1993-12-31

    Aneuploidy is a relatively common genetic disorder that results in human morbidity and mortality. Approximately 30% of embryonic and fetal deaths and 3.45 per thousand livebirths are associated with an abnormal number of chromosomes. Unfortunately, very little is known about the etiology and mechanism of chromosome missegregation. This situation dictates that considerable research be directed toward understanding the causes of aneuploidy. Although several hypotheses have been advanced for the etiology of aneuploidy, there still exists a paucity of information about the direct cuases and mechanisms of aneuploidy production. Without such specific knowledge, there is little hope of reducing the incidence of aneuploidy in humans. Some progress has been made. We now know that various chemicals can induce aneuploidy by interacting with certain cellular organelles, especially components of the spindle apparatus. These results have been demonstrated in various organisms and cell types both in vivo and in vitro. Since the ultimate objective of aneuploidy research is to obtain information that can be used to reduce the aneuploidy burden in humans, we have concentrated our research efforts on studying chemically-induced aneuploidy in mammalian germ cells and zygotes.

  11. Preparation of intact chloroplasts by chemically induced lysis of the green alga Dunaliella marina.

    Science.gov (United States)

    Kombrink, E; Wöber, G

    1980-07-01

    A method for the isolation in high yield of intact chloroplasts from the unicellular green alga Dunaliella marina (Volvocales) is described. This procedure uses chemically induced lysis of cells with the polycationic macromolecules, DEAE-dextran (M=500,000) or poly-D,L-lysine (M=30,000-70,000). Reaction conditions were optimized with respect to obtaining a high yield of intact chloroplasts, after isopycnic centrifugation in a linear sucrose density gradient, by varying the concentration of polycation and the temperature and pH of incubation. Broken chloroplasts devoid of the stromal marker enzymes fructosebisphosphate phosphatase and ribulosebisphosphate carboxylase, but containing mitochondrial (fumarase) and microbody (catalase) contamination, were banded at a bouyant density of 1.18 g cm(-3). Intact chloroplasts, as indicated by their retention of alkaline fructosebisphosphate phosphatase and ribulosebisphosphate carboxylase, were found in 30% yield (chlorophyll in intact cells, 100%) at an equilibrium density of 1.24 g cm(-3). Contamination by cytoplasmic material (pyruvate kinase), mitochondria, and microbodies was less than 8% each. PMID:24306242

  12. Extraction of chemical-induced diseases using prior knowledge and textual information.

    Science.gov (United States)

    Pons, Ewoud; Becker, Benedikt F H; Akhondi, Saber A; Afzal, Zubair; van Mulligen, Erik M; Kors, Jan A

    2016-01-01

    We describe our approach to the chemical-disease relation (CDR) task in the BioCreative V challenge. The CDR task consists of two subtasks: automatic disease-named entity recognition and normalization (DNER), and extraction of chemical-induced diseases (CIDs) from Medline abstracts. For the DNER subtask, we used our concept recognition tool Peregrine, in combination with several optimization steps. For the CID subtask, our system, which we named RELigator, was trained on a rich feature set, comprising features derived from a graph database containing prior knowledge about chemicals and diseases, and linguistic and statistical features derived from the abstracts in the CDR training corpus. We describe the systems that were developed and present evaluation results for both subtasks on the CDR test set. For DNER, our Peregrine system reached anF-score of 0.757. For CID, the system achieved anF-score of 0.526, which ranked second among 18 participating teams. Several post-challenge modifications of the systems resulted in substantially improvedF-scores (0.828 for DNER and 0.602 for CID). RELigator is available as a web service athttp://biosemantics.org/index.php/software/religator. PMID:27081155

  13. Adenovirus-mediated human brain-derived neurotrophic factor gene-modified bone marrow mesenchymal stem cell transplantation for spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    Changsheng Wang; Jianhua Lin; Chaoyang Wu; Rongsheng Chen

    2011-01-01

    Rat bone marrow mesenchymal stem cells expressing brain-derived neurotrophic factor were successfully obtained using a gene transfection method, then intravenously transplanted into rats with spinal cord injury. At 1, 3, and 5 weeks after transplantation, the expression of ??brain-derived neurotrophic factor and neurofilament-200 was upregulated in the injured spinal cord, spinal cord injury was alleviated, and Basso-Beattie-Bresnahan scores of hindlimb motor function were significantly increased. This evidence suggested that intravenous transplantation of adenovirus- mediated brain-derived neurotrophic factor gene-modified rat bone marrow mesenchymal stem cells could play a dual role, simultaneously providing neural stem cells and neurotrophic factors.

  14. 骨髓间充质干细胞移植促进糖尿病大鼠β细胞再生的机制%Mechanism of bone marrow mesenchymal stem cells transplantation in promoting β-cell regeneration in diabetic rats

    Institute of Scientific and Technical Information of China (English)

    2013-01-01

      目的探索骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)移植促进糖尿病大鼠模型中β细胞再生的机制。方法正常SD大鼠给予单次大剂量链脲佐菌素(streptozotocin,STZ)(65 mg/kg)注射,3 d后开始监测血糖,连续3 d随机血糖>28 mmol/L者选为实验对象。所有实验对象随机分为对照组(n=15),BMSC移植组(n=15),BMSC培养上清移植组(n=15)以及BMSC示踪组(以Dil染料标记BMSCs,n=5),分别接受PBS、MSC、MSC培养上清及Dil-BMSC的移植。结果 BMSC移植后30 d左右血糖水平有明显改善[(20.4±0.98) mmol/L,n=15],胰岛素水平有明显提高[(0.99±0.23) ng/ml, n=15],β细胞质量明显增长。富含BMSC分泌因子的BMSC上清输注后,糖尿病大鼠血糖有一定程度的降低[(26.2±1.53) mmol/L, n=15],胰岛素水平有所恢复[(0.341±0.08) ng/ml,n=15],β细胞质量较对照组有所增加。在胰腺仅检测到小部分BMSC,这部分归巢于胰腺的BMSC中检测不到与胰岛素的共标。结论 BMSC的分泌效应为BMSC促进糖尿病动物模型中β细胞再生的机制。%Objective To study the mechanism of bone marrow mesenchymal stem cells (BMSC) transplantation in promotingβ-cell regeneration in diabetic model of SD rats. Methods Three days after a single high streptozotocin (STZ) dose (65 mg/kg) was injected into normal SD rats, their blood glucose levels were measured. Those with their blood glucose level>28 mmol/L for 3 consecutive days were randomly divided into control group (n=15), BMSC transplantation group (n=15), BMSC supernatant transplantation group (n=15), and BMSC (marked with Dil stain) trace group (n=5). Results The blood glucose level was significantly lower (20.4 ±0.98 mmol/L, 26.2±1.53 mmol/L), the serum insulin level was significantly higher (0.99±0.23 ng/ml, 0.341±0.08 ng/ml), and theβcells regenerated faster in BMSC transplantation group and BMSC supernatant transplantation group

  15. bFGF促进大鼠骨髓间充质干细胞向神经胶质样细胞转分化%bTGF Promotes the Transdifferentiation of Mesenchymal Stem Cells into Neuroglia-like Cells in Rat

    Institute of Scientific and Technical Information of China (English)

    陈莉; 买霞; 扎拉嘎胡; 陈小义; 徐瑞成

    2008-01-01

    目的 研究大鼠骨髓间充质干细胞(rat mesenchymal stem cells,rMSCs)在体外诱导为神经源性细胞的可行性,并探讨其作用机制. 方法从Wistar大鼠骨髓中获得rMSCs.纯化培养传代后用不同浓度的bFGF诱导,经MTT法检测bFGF对rMSCs生长的影响;用倒置光显微镜、透射电镜、免疫组化和RT-PCR等方法鉴定诱导后细胞行为改变与snail mRNA表达.结果 bFGF对rMSCs具有促增殖作用,在倒置光显微镜和透射电镜下可见到有神经胶质样细胞形成, 免疫组化证明GFAP的表达较对照组显著增高(P<0.01).RT-PCR结果表明,诱导组细胞snail mRNA明显表达.结论 bFGF促进rMSCs转分化为GFAP阳性的神经胶质样细胞,转录因子Snail可能与转化过程有关.

  16. Bone marrow mesenchymal stem cells with Nogo-66 receptor gene silencing for repair of spinal cord injur y

    Institute of Scientific and Technical Information of China (English)

    Zhiyuan Li; Zhanxiu Zhang; Lili Zhao; Hui Li; Suxia Wang; Yong Shen

    2014-01-01

    We hypothesized that RNA interference to silence Nogo-66 receptor gene expression in bone marrow mesenchymal stem cells before transplantation might further improve neurological function in rats with spinal cord transection injury. After 2 weeks, the number of neurons and BrdU-positive cells in the Nogo-66 receptor gene silencing group was higher than in the bone marrow mesenchymal stem cell group, and significantly greater compared with the model group. After 4 weeks, behavioral performance was signiifcantly enhanced in the model group. Af-ter 8 weeks, the number of horseradish peroxidase-labeled nerve ifbers was higher in the Nogo-66 receptor gene silencing group than in the bone marrow mesenchymal stem cell group, and signiifcantly higher than in the model group. The newly formed nerve ifbers and myelinated ner ve ifbers were detectable in the central transverse plane section in the bone marrow mesenchymal stem cell group and in the Nogo-66 receptor gene silencing group.

  17. Transplantation of bone marrow mesenchymal stem cells to improve blood glucose and urinary total protein levels in diabetic nephropathy rats%骨髓间充质干细胞移植改善糖尿病肾病大鼠血糖及尿总蛋白

    Institute of Scientific and Technical Information of China (English)

    杜俊文; 吴韬; 张坤; 苏白玉; 卢彩萍; 王伟超; 雷琳; 郭静霞

    2016-01-01

    BACKGROUND:Common strategies for preventing diabetic nephropathy include effective control of blood sugar and blood pressure, inhibition of the rennin-angiotensin system and lipid-lowering therapy, but it is often difficult to get the desired results. OBJECTIVE:To investigate the effect of transplantation of bone marrow mesenchymal stem cels on levels of blood glucose and urinary total protein in diabetic nephropathy rats. METHODS: Forty-five Sprague-Dawley rats were randomly divided into three groups (n=15 per group): normal control group, diabetic nephropathy group and stem cel transplantation group. Rats in the diabetic nephropathy and stem cel transplantation groups were given single use of 60 mg/kg streptozotocin to make diabetic nephropathy models. The same dose of citric acid-sodium citrate buffer was injected in the normal control group. After modeling, 200μL of bone marrow mesenchymal stem cel solution (2×106) was injected into the left ventricle of rats in the stem cel transplantation group, and then at 7 days after the first transplantation, the cel transplantation was conducted again. The same dose of serum-free L-DMEM was injected intracardialy into the rats in the normal control and diabetic nephropathy groups. Levels of urinary total protein and blood glucose were detected. RESULTS AND CONCLUSION:At 1, 4, 8 weeks after treatment, the urinary total protein and blood glucose levels were significantly higher in the stem cel transplantation group and diabetic nephropathy group than the normal control group (P 0.05). These findings indicate that bone marrow mesenchymal stem cel transplantation in diabetic nephropathy rats can get good results in a short period, significantly improve the blood glucose and urinary total protein levels, but the long-term treatment effect is poor.%背景:糖尿病肾病防治的基本措施包括有效控制血糖、控制血压、抑制肾素-血管紧张素系统和调脂治疗等,但往往无法获得理想的效果

  18. Role of Diasulin, an Herbal Formulation on Antioxidant Status in Chemical Induced Diabetes

    Directory of Open Access Journals (Sweden)

    Leelavinothan Pari

    2006-01-01

    Full Text Available In the present study the effect of ethanolic extract of the Diasulin was examined for its antioxidant effect in the liver and kidney of alloxan diabetic rats. Oral administration of Diasulin (200 mg kg-1 for 30 days resulted in a significant reduction in thiobarbituric acid reactive substances and hydroperoxides. The Diasulin also caused a significant increase in reduced glutathione, superoxide dismutase, catalase, glutathione peroxidase and glutathione-S-transferase in liver and kidney of alloxan diabetic rats. The results show the antioxidant effect of Diasulin. The effect of Diasulin at 200 mg kg-1 was more effective than glibenclamide, a reference drug.

  19. Evaluation of anticonvulsant activity of volatile oil extract of Nigella sativa seeds by chemically induced seizure model in albino rats

    Directory of Open Access Journals (Sweden)

    Asmatanzeem Bepari

    2016-08-01

    Conclusions: The N. sativa seeds showed anticonvulsant activity in pentylenetetrazole induced seizure model of epilepsy. This study showed that volatile oil of N. sativa seeds potentiated the effect of sodium valproate. [Int J Basic Clin Pharmacol 2016; 5(4.000: 1300-1307

  20. Overexpression of microRNA-124 promotes the neuronal differentiation of bone marrow-derived mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    Defeng Zou; Yi Chen; Yaxin Han; Chen Lv; Guanjun Tu

    2014-01-01

    microRNAs (miRNAs) play an important regulatory role in the self-renewal and differentiation of stem cells. In this study, we examined the effects of miRNA-124 (miR-124) overexpression in bone marrow-derived mesenchymal stem cells. In particular, we focused on the effect of overexpression on the differentiation of bone marrow-derived mesenchymal stem cells into neurons. First, we used GeneChip technology to analyze the expression of miRNAs inbone marrow-derived mesen-chymal stem cells, neural stem cells and neurons. miR-124 expression was substantially reduced inbone marrow-derived mesenchymal stem cells compared with the other cell types. We con-structed a lentiviral vector overexpressing miR-124 and transfected it intobone marrow-derived mesenchymal stem cells. Intracellular expression levels of the neuronal early markersβ-III tu-bulin and microtubule-associated protein-2 were signiifcantly increased, and apoptosis induced by oxygen and glucose deprivation was reduced in transfected cells. After miR-124-transfected bone marrow-derived mesenchymal stem cells were transplanted into the injured rat spinal cord, a large number of cells positive for the neuronal marker neurofilament-200 were observed in the transplanted region. The Basso-Beattie-Bresnahan locomotion scores showed that the motor function of the hind limb of rats with spinal cord injury was substantially improved. These re-sults suggest that miR-124 plays an important role in the differentiation ofbone marrow-derived mesenchymal stem cells into neurons. Our ifndings should facilitate the development of novel strategies for enhancing the therapeutic efifcacy ofbone marrow-derived mesenchymal stem cell transplantation for spinal cord injury.

  1. 660 nm red light-enhanced bone marrow mesenchymal stem cell transplantation for hypoxic-ischemic brain damage treatment

    Institute of Scientific and Technical Information of China (English)

    Xianchao Li; Wensheng Hou; Xiaoying Wu; Wei Jiang; Haiyan Chen; Nong Xiao; Ping Zhou

    2014-01-01

    Bone marrow mesenchymal stem cell transplantation is an effective treatment for neonatal hy-poxic-ischemic brain damage. However, the in vivo transplantation effects are poor and their survival, colonization and differentiation efifciencies are relatively low. Red or near-infrared light from 600-1,000 nm promotes cellular migration and prevents apoptosis. Thus, we hypothesized that the combination of red light with bone marrow mesenchymal stem cell transplantation would be effective for the treatment of hypoxic-ischemic brain damage. In this study, the migra-tion and colonization of cultured bone marrow mesenchymal stem cells on primary neurons after oxygen-glucose deprivation were detected using Transwell assay. The results showed that, after a 40-hour irradiation under red light-emitting diodes at 660 nm and 60 mW/cm2, an increasing number of green lfuorescence-labeled bone marrow mesenchymal stem cells migrated towards hypoxic-ischemic damaged primary neurons. Meanwhile, neonatal rats with hypoxic-ischemic brain damage were given an intraperitoneal injection of 1 × 106 bone marrow mesenchymal stem cells, followed by irradiation under red light-emitting diodes at 660 nm and 60 mW/cm2 for 7 successive days. Shuttle box test results showed that, after phototherapy and bone marrow mesenchymal stem cell transplantation, the active avoidance response rate of hypoxic-ischemic brain damage rats was significantly increased, which was higher than that after bone marrow mesenchymal stem cell transplantation alone. Experimental ifndings indicate that 660 nm red light emitting diode irradiation promotes the migration of bone marrow mesenchymal stem cells, thereby enhancing the contribution of cell transplantation in the treatment of hypox-ic-ischemic brain damage.

  2. Selenium and vitamin E inhibit radiogenic and chemically induced transformation in vitro via different mechanisms

    International Nuclear Information System (INIS)

    Results from in vivo and in vitro studies showing that antioxidants may act as anticarcinogens support the role of active oxygen in carcinogenesis and provide impetus for exploring the functions of dietary antioxidants in cancer prevention by using in vitro models. The authors examined the single and combined effects of selenium, a component of glutathione peroxidase, and vitamin E, a known antioxidant, on cell transformation induced in C3H/10T-1/2 cells by x-rays, benzo[a]pyrene, or tryptophan pyrolysate and on the levels of cellular scavenging systems peroxide destruction. Incubation of C3H/10T-1/2 cells with 2.5 μM Na2SeO3 (selenium) or with 7 μM α-tocopherol succinate (vitamin E) 24 hr prior to exposure to x-rays or the chemical carcinogens resulted in an inhibition of transformation by each of the antioxidants with an additive-inhibitory action when the two nutrients were combined. Cellular pretreatment with selenium resulted in increased levels of cellular glutathione peroxidase, catalase, and nonprotein thiols (glutathione) and in an enhanced destruction of peroxide. The results support our earlier studies showing that free radical-mediated events play a role in radiation and chemically induced transformation. They indicate that selenium and vitamin E act alone and in additive fashion as radioprotecting and chemopreventing agents. The results further suggest that selenium confers protection in part by inducing or activating cellular free-radical scavenging systems and by enhancing peroxide breakdown while vitamin E appears to confer its protection by and alternate complementary mechanism

  3. Selenium and vitamin E inhibit radiogenic and chemically induced transformation in vitro via different mechanisms

    Energy Technology Data Exchange (ETDEWEB)

    Borek, C.; Ong, A.; Mason, H.; Donahue, L.; Biaglow, J.E.

    1986-03-01

    Results from in vivo and in vitro studies showing that antioxidants may act as anticarcinogens support the role of active oxygen in carcinogenesis and provide impetus for exploring the functions of dietary antioxidants in cancer prevention by using in vitro models. The authors examined the single and combined effects of selenium, a component of glutathione peroxidase, and vitamin E, a known antioxidant, on cell transformation induced in C3H/10T-1/2 cells by x-rays, benzo(a)pyrene, or tryptophan pyrolysate and on the levels of cellular scavenging systems peroxide destruction. Incubation of C3H/10T-1/2 cells with 2.5 ..mu..M Na/sup 2/SeO/sup 3/ (selenium) or with 7 ..mu..M ..cap alpha..-tocopherol succinate (vitamin E) 24 hr prior to exposure to x-rays or the chemical carcinogens resulted in an inhibition of transformation by each of the antioxidants with an additive-inhibitory action when the two nutrients were combined. Cellular pretreatment with selenium resulted in increased levels of cellular glutathione peroxidase, catalase, and nonprotein thiols (glutathione) and in an enhanced destruction of peroxide. The results support our earlier studies showing that free radical-mediated events play a role in radiation and chemically induced transformation. They indicate that selenium and vitamin E act alone and in additive fashion as radioprotecting and chemopreventing agents. The results further suggest that selenium confers protection in part by inducing or activating cellular free-radical scavenging systems and by enhancing peroxide breakdown while vitamin E appears to confer its protection by and alternate complementary mechanism.

  4. Transplantation of mesenchymal stem cells improves type 1 diabetes mellitus.

    Science.gov (United States)

    Li, Lisha; Li, Furong; Gao, Feng; Yang, Yali; Liu, Yuanyuan; Guo, Pingping; Li, Yulin

    2016-05-01

    Bone-marrow-derived stem cells can regenerate pancreatic tissue in a model of type 1 diabetes mellitus. Mesenchymal stem cells (MSCs) form the main part of bone marrow. We show that the intrapancreatic transplantation of MSCs elevates serum insulin and C-peptide, while decreasing blood glucose. MSCs engrafted into the damaged rat pancreas become distributed into the blood vessels, acini, ducts, and islets. Renascent islets, islet-like clusters, and a small number of MSCs expressing insulin protein have been observed in the pancreas of diabetic rats. Intrapancreatic transplantation of MSCs triggers a series of molecular and cellular events, including differentiation towards the pancreas directly and the provision of a niche to start endogenous pancreatic regeneration, which ameliorates hypoinsulinemia and hyperglycemia caused by streptozotocin. These data establish the many roles of MSCs in the restoration of the function of an injured organ. PMID:26650464

  5. Exposure to Gulf War Illness chemicals induces functional muscarinic receptor maladaptations in muscle nociceptors.

    Science.gov (United States)

    Cooper, B Y; Johnson, R D; Nutter, T J

    2016-05-01

    Chronic pain is a component of the multisymptom disease known as Gulf War Illness (GWI). There is evidence that pain symptoms could have been a consequence of prolonged and/or excessive exposure to anticholinesterases and other GW chemicals. We previously reported that rats exposed, for 8 weeks, to a mixture of anticholinesterases (pyridostigmine bromide, chlorpyrifos) and a Nav (voltage activated Na(+) channel) deactivation-inhibiting pyrethroid, permethrin, exhibited a behavior pattern that was consistent with a delayed myalgia. This myalgia-like behavior was accompanied by persistent changes to Kv (voltage activated K(+)) channel physiology in muscle nociceptors (Kv7, KDR). In the present study, we examined how exposure to the above agents altered the reactivity of Kv channels to a muscarinic receptor (mAChR) agonist (oxotremorine-M). Comparisons between muscle nociceptors harvested from vehicle and GW chemical-exposed rats revealed that mAChR suppression of Kv7 activity was enhanced in exposed rats. Yet in these same muscle nociceptors, a Stromatoxin-insensitive component of the KDR (voltage activated delayed rectifier K(+) channel) exhibited decreased sensitivity to activation of mAChR. We have previously shown that a unique mAChR-induced depolarization and burst discharge (MDBD) was exaggerated in muscle nociceptors of rats exposed to GW chemicals. We now provide evidence that both muscle and vascular nociceptors of naïve rats exhibit MDBD. Examination of the molecular basis of the MDBD in naïve animals revealed that while the mAChR depolarization was independent of Kv7, the action potential burst was modulated by Kv7 status. mAChR depolarizations were shown to be dependent, in part, on TRPA1. We argue that dysfunction of the MDBD could be a functional convergence point for maladapted ion channels and receptors consequent to exposure to GW chemicals. PMID:27058124

  6. Combining acellular nerve allografts with brain-derived neurotrophic factor transfected bone marrow mesenchymal stem cells restores sciatic nerve injury better than either intervention alone

    OpenAIRE

    Zhang, Yanru; Zhang, Hui; Zhang, Gechen; Ka, Ka; Huang, Wenhua

    2014-01-01

    In this study, we chemically extracted acellular nerve allografts from bilateral sciatic nerves, and repaired 10-mm sciatic nerve defects in rats using these grafts and brain-derived neurotrophic factor transfected bone marrow mesenchymal stem cells. Experiments were performed in three groups: the acellular nerve allograft bridging group, acellular nerve allograft + bone marrow mesenchymal stem cells group, and the acellular nerve allograft + brain-derived neurotrophic factor transfected bone...

  7. 大鼠骨髓间充质干细胞培养、鉴定与成胰岛样β细胞诱导%Cultivation, Identification and Islet β Cells Induction of Bone Marrow Mesenchymal Stem Cells in Rats

    Institute of Scientific and Technical Information of China (English)

    王鹤霏; 任宇; 呼啸; 张飞旭; 李晓聪; 仓明; 刘东军

    2016-01-01

    To find new substitution of islet β cells for treating diabetes,rat bone marrow mesenchymal stem cells (BMSCs) were isolated,cultured,identified and induced into islet β cells in vitro.Isolation of rat BMSCs by the whole bone marrow centrifugation and adherence method,then BMSCs were cultured,subcultured,detected cell surface markers and induced into islet β cells by DMSO and high glucose environment.The results showed that the morphology of BMSCs was fibroblast like,with active passage ability.CD 13,CD44 and CD 106 antigens were detected,but CD49d antigen was negative.Under the condition of islet β cells induction,BMSCs could form isletlike round cell mass,dithizone staining was brownish red,PDX1,CK19,nestin and insulin antigens were positive using immunohistochemistry staining.BMSCs induction into islet β cells expressed insulin,PDX1 and glucagon detected by RT-PCR.The expression quantity of insulin mRNA in differentiated BMSCs was 1.09 times (P>0.05) than that of in pancreas adult stem cells (PASCs) detected by q-PCR.At the same time,differentiated BMSCs had the corresponding expression of insulin protein.The results demonstrated that rat BMSCs grow stably in vitro,and can be converted into islet-like cells,providing new experimental basis for diabetes treatment.%为治疗糖尿病寻找新的胰岛β细胞替代物,该研究对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)进行了体外分离培养、鉴定及成胰岛样β细胞诱导.应用全骨髓离心贴壁培养法分离大鼠BMSCs,进行培养、传代、表面标志物检测,利用DMSO和高糖环境对BMSCs进行胰岛样β细胞诱导.结果显示,大鼠BMSCs体外培养细胞形态呈成纤维细胞样,可以稳定传代;CD13、CD44和CD106表达呈阳性,CD49d呈阴性.在成胰诱导条件下,BMSCs可形成胰岛样圆形细胞团,双硫腙染色呈棕红色,PDX1(pancreatic duodenal homeodomain 1)、CK19(cytokeratin 19)、巢蛋白、胰岛素免疫组化染色

  8. Combination of high performance refractometry and infrared spectroscopy as a probe for chemically induced gelation and vitrification of epoxies

    OpenAIRE

    Müller, Ulrich; Philipp, Martine; Gervais, P. C.; Possart, Prof Dr Wulff; Wehlack, C.; Kieffer, J.; Sanctuary, Roland; Krüger, Jan-Kristian

    2010-01-01

    A combination of infrared spectroscopy and high performance refractometry was used to investigate the chemically induced sol-gel and glass transition during the polymerization of epoxies. Representations of the refractive index versus chemical conversion reveal an interesting insight in the optical properties accompanying gelation and vitrification. Whereas the electronic polarizability of the liquid state of small average molecular mass and the glassy state is dominated by the mass density, ...

  9. Exact Solution to the Extended Zwanzig Model for Quasi-Sigmoidal Chemically Induced Denaturation Profiles: Specific Heat and Configurational Entropy

    OpenAIRE

    Aguilar-Pineda, G. E.; Olivares-Quiroz, L.

    2014-01-01

    Temperature and chemically induced denaturation comprise two of the most characteristic mechanisms to achieve the passage from the native state N to any of the unstructured states Dj in the denatured ensemble in proteins and peptides. In this work we present a full analytical solution for the configurational partition function qs of a homopolymer chain poly-X in the extended Zwanzig model (EZM) for a quasisigmoidal denaturation profile. This solution is built up from an EZM exact solution in...

  10. Role of the Slug Transcription Factor in Chemically-Induced Skin Cancer

    Directory of Open Access Journals (Sweden)

    Kristine von Maltzan

    2016-02-01

    Full Text Available The Slug transcription factor plays an important role in ultraviolet radiation (UVR-induced skin carcinogenesis, particularly in the epithelial-mesenchymal transition (EMT occurring during tumor progression. In the present studies, we investigated the role of Slug in two-stage chemical skin carcinogenesis. Slug and the related transcription factor Snail were expressed at high levels in skin tumors induced by 7,12-dimethylbenz[α]anthracene application followed by 12-O-tetradecanoylphorbol-13-acetate (TPA treatment. TPA-induced transient elevation of Slug and Snail proteins in normal mouse epidermis and studies in Slug transgenic mice indicated that Slug modulates TPA-induced epidermal hyperplasia and cutaneous inflammation. Although Snail family factors have been linked to inflammation via interactions with the cyclooxygenase-2 (COX-2 pathway, a pathway that also plays an important role in skin carcinogenesis, transient TPA induction of Slug and Snail appeared unrelated to COX-2 expression. In cultured human keratinocytes, TPA induced Snail mRNA expression while suppressing Slug expression, and this differential regulation was due specifically to activation of the TPA receptor. These studies show that Slug and Snail exhibit similar patterns of expression during both UVR and chemical skin carcinogenesis, that Slug and Snail can be differentially regulated under some conditions and that in vitro findings may not recapitulate in vivo results.

  11. Improvement of learning and memory impairment of Alzheimer's disease rat models treated by bone mesenchymal stem cells modified with BDNF gene%BDNF基因修饰骨髓间充质干细胞对AD大鼠认知障碍的改善

    Institute of Scientific and Technical Information of China (English)

    张平; 赵钢勇; 陈凯; 张杏红; 苏立凯

    2011-01-01

    目的:研究经侧脑室移植脑源性神经生长因子(BDNF)修饰骨髓间充质干细胞(MSCs)对AD大鼠认知功能的改善作用.方法:采用双侧海马注射Aβ25-35制备AD大鼠模型,筛选40只雄性SD大鼠随机分为5组,假手术组(Sham组),AD模型组(AD组),移植骨髓MSCs治疗组(MSCs组),移植pIRESneo转染的骨髓MSCs治疗组(CON组)和移植pIRESneo-EGFP-BDNF转染的骨髓MSCs组(BDNF组),每组各8只.采用Morris水迷宫测试大鼠的学习记忆能力.结果:AD组与Sham组大鼠比较平均逃避潜伏期(AEL)延长、跨越平台次数明显减少(P<0.01),搜索策略明显差(P <0.01);MSCs组、CON组、BDNF组与AD组大鼠比较AEL明显缩短、跨越平台次数显著增加(P <0.05或P<0.01),搜索策略显著好转(P<0.01),且BDNF组与CON组、MSCs组大鼠比较改善更显著(P<0.05或P<0.01).结论:经侧脑室移植BDNF修饰的骨髓MSCs可以显著改善AD大鼠的学习认知能力.%Objective: To investigate the improvement of learning and memory of Alzheimer's disease ( AD) in rat model treated by brain-derived neurotrophic factor (BDNF) gene modified bone mesenchymal stem cells (MSCs) in jected through cerebral lateral ventricle. Methods; The AD rat models were constructed by injection of beta-amyloid peptide 25-35 (Aβ25_35) into the hippocampus on both sides stereotaxically. Forty healthy male SD rats were selected and then randomly divided into five groups of eight each, I. E. Sham operation group (Sham group) , AD rat model group (AD group) , bone MSCs group (MSCs group) , bone MSCs modified with plRESneo plasmids group (CON group) and bone MSCs modified with pIRESneo-EGFP-BDNF plasmids group ( BDNF group). The learning and memory abilities of AD rat models were evaluated with Morris water maze test. Results; The average escape latency (AEL) was significantly prolonged and the frequency of crossing over the platform was significantly decreased and the searching strategy was poorer in AD

  12. Expression of Odontogenic Genes in Human Bone Marrow Mesenchymal Stem Cells

    Directory of Open Access Journals (Sweden)

    Seyedeh Sara Bagheri

    2013-01-01

    Full Text Available Objective: Tooth loss is a common problem and since current tooth replacement methods cannot counter balance with biological tooth structures, regenerating natural tooth structures has become an ideal goal. A challenging problem in tooth regeneration is to find a proper clinically feasible cell to seed.This study was designed to investigate the odontogenic potential of human bone marrow mesenchymal stem cells (HBMSCs for seeding in tooth regeneration.Materials and Methods: In this experimental study, three pregnant Sprague Dawley (SD rats were used at the eleventh embryonic day and rat fetuses were removed surgically using semilunar flap under general anesthesia. The primary mandible was cut using a stereomicroscope. The epithelial and mesenchymal components were separated and the dissected oral epithelium was cultured for 3 days. We used flow cytometry analysis to confirm presence of mesenchymal stem cells and not hematopoietic cells and to demonstrate the presence of oral epithelium. Bone marrow mesenchymal stem cells (BMSCs and cultured oral epithelium were then co-cultured for 14 days. BMSCs cultured alone were used as controls. Expression of two odontogenic genes Pax9 and DMP1 was assessed using quantitative reverse transcription- polymerase chain reaction (RT-PCR.Results: Expression of two odontogenic genes, Pax9 and DMP1, were detected in BMSCs co-cultured with oral epithelium but not in the control group.Conclusion: Expression of Pax9 and DMP1 by human BMSCs in the proximity of odontogenic epithelium indicates odontogenic potential of these cells.

  13. Pattern formation by vascular mesenchymal cells

    OpenAIRE

    Garfinkel, Alan; Tintut, Yin; Petrasek, Danny; Boström, Kristina; Demer, Linda L.

    2004-01-01

    In embryogenesis, immature mesenchymal cells aggregate and organize into patterned tissues. Later in life, a pathological recapitulation of this process takes place in atherosclerotic lesions, when vascular mesenchymal cells organize into trabecular bone tissue within the artery wall. Here we show that multipotential adult vascular mesenchymal cells self-organize in vitro into patterns that are predicted by a mathematical model based on molecular morphogens interacting in a reaction-diffusion...

  14. B-lymphocytes as key players in chemical-induced asthma.

    Directory of Open Access Journals (Sweden)

    Vanessa De Vooght

    Full Text Available T-lymphocytes and B-lymphocytes are key players in allergic asthma, with B-lymphocytes producing antigen-specific immunoglobulins E (IgE. We used a mouse model of chemical-induced asthma and transferred B-lymphocytes from sensitized animals into naïve wild type mice, B-lymphocyte knock-out (B-KO mice or severe combined immunodeficiency (SCID mice. On days 1 and 8, BALB/c mice were dermally sensitized with 0.3% toluene diisocyanate (TDI (20 µl/ear. On day 15, mice were euthanized and the auricular lymph nodes isolated. B-lymphocytes (CD19(+ were separated from the whole cell suspension and 175,000 cells were injected in the tail vein of naïve wild type, B-KO or SCID mice. Three days later, the mice received a single oropharyngeal challenge with 0.01% TDI (20 µl or vehicle (acetone/olive oil (AOO (controls. Airway reactivity to methacholine and total and differential cell counts in the bronchoalveolar lavage (BAL fluid were measured 24 hours after challenge. B-lymphocytes of AOO or TDI-sensitized mice were characterized for the expression of surface markers and production of cytokines. We found that transfer of B-cells obtained from mice dermally sensitized to toluene diisocyanate (TDI into naïve wild type mice, B-KO mice or SCID mice led, within three days, to an acute asthma-like phenotype after an airway challenge with TDI. This response was specific and independent of IgE. These B-lymphocytes showed antigen presenting capacities (CD80/CD86 and CD40 and consisted of B effector (Be2- (IL-4 and Be1-lymphocytes (IFN-γ. The transferred B-lymphocytes were visualized near large airways, 24 hours after TDI challenge. Thus, B-lymphocytes can provoke an asthmatic response without the action of T-lymphocytes and without major involvement of IgE.

  15. Trypanosoma cruzi extracts elicit protective immune response against chemically induced colon and mammary cancers.

    Science.gov (United States)

    Ubillos, Luis; Freire, Teresa; Berriel, Edgardo; Chiribao, María Laura; Chiale, Carolina; Festari, María Florencia; Medeiros, Andrea; Mazal, Daniel; Rondán, Mariella; Bollati-Fogolín, Mariela; Rabinovich, Gabriel A; Robello, Carlos; Osinaga, Eduardo

    2016-04-01

    Trypanosoma cruzi, the protozoan parasite that causes Chagas' disease, has anticancer effects mediated, at least in part, by parasite-derived products which inhibit growth of tumor cells. We investigated whether immunity to T. cruzi antigens could induce antitumor activity, using two rat models which reproduce human carcinogenesis: colon cancer induced by 1,2-dimethylhydrazine (DMH), and mammary cancer induced by N-nitroso-N-methylurea (NMU). We found that vaccination with T. cruzi epimastigote lysates strongly inhibits tumor development in both animal models. Rats immunized with T. cruzi antigens induce activation of both CD4(+) and CD8(+) T cells and splenocytes from these animals showed higher cytotoxic responses against tumors as compared to rats receiving adjuvant alone. Tumor-associated immune responses included increasing number of CD11b/c(+) His48(-) MHC II(+) cells corresponding to macrophages and/or dendritic cells, which exhibited augmented NADPH-oxidase activity. We also found that T. cruzi lysate vaccination developed antibodies specific for colon and mammary rat cancer cells, which were capable of mediating antibody-dependent cellular cytotoxicity (ADCC) in vitro. Anti-T. cruzi antibodies cross-reacted with human colon and breast cancer cell lines and recognized 41/60 (68%) colon cancer and 38/63 (60%) breast cancer samples in a series of 123 human tumors. Our results suggest that T. cruzi antigens can evoke an integrated antitumor response involving both the cellular and humoral components of the immune response and provide novel insights into the understanding of the intricate relationship between parasite infection and tumor growth. PMID:26519949

  16. Chemopreventive effect of Quercus infectoria against chemically induced renal toxicity and carcinogenesis

    OpenAIRE

    Rehman, Muneeb U.; Mir Tahir, Farrah Ali; Wajhul Qamar; Rehan Khan; Abdul Quaiyoom Khan; Abdul Lateef; Oday-O-Hamiza; Sarwat Sultana

    2012-01-01

    In this study we have shown that Quercus infectoria attenuates Fe- NTA induced renal oxidative stress, hyperproliferative response and renal carcinogenesis in rats. Fe-NTA promoted DEN (N-diethyl nitrosamine) initiated renal carcinogenesis by increasing the percentage incidence of tumors and induces early tumor markers viz. ornithine decarboxylase (ODC) level and PCNA expression. Fe- NTA (9 mg Fe/kg body weight, intraperitoneally) enhances renal Malondialdehyde, xanthine oxidase and hydrogen ...

  17. 人参皂苷Rg1联合骨髓间充质干细胞移植对痴呆大鼠学习记忆能力的影响%Effect of Ginsenoside Rg1 on the Spatial Learning-memory Ability in Dementia Rats after Transplanted with Bone Marrow Mesenchymal Stem Cells

    Institute of Scientific and Technical Information of China (English)

    邬伟; 杨景全; 何志勇; 王小同

    2011-01-01

    Objective To study the effect and mechanism of ginsenoside Rg1 on the spatial learning-memory ability in rats with Alzheimer's disease after transplanted with bone marrow mesenchymal stem cells (BMSCs).Methods Using digital randomization table method, seventy-five male SD rats were divided into the bilateral FF transection model group (as the model group: ambi-hippocampal fimbria-fomix transected), the sham-operative control group (the SOC group: receiving the same modeling process as the model group, but without ambi-hippocampal fimbria-fornix transected), the ginsenoside Rg1 treatment group (as the treatment group: Two weeks after modeling ginsenoside Rg1 was peritoneally injected at the dose of 5 mg/kg, once daily for four weeks in total), the BMSCs transplanted treatment group [as the control group: Two weeks after modeling every rat received transplantation of BMSCs (10 μL, 1 ×106cells)], and the ginsenoside Rg1 +BMSCs treatment group (as the combination group: They received both transplantation of BMSCs and peritoneal injection of ginsenoside Rg1 ).The spatial learning-memory ability of rats was detected by Morris water maze and the escape latency (s) was recorded.mRNA expression of nerve growth factor (NGF) was detected using Real-time PCR.Results Six weeks after the hippocampal fimbria-fornix (FF) transection,the escape latency of each medication group was obviously shorter than that of the model group, and the spatial learning-memory ability of dementia rats was somewhat improved.The spatial learning-memory ability of rats in the combination group was (29.95 ±2.03) and the mRNA expression level of NGF was (1.13 ±0.15), better than those in the BMSCs group (44.36 ±1.43, 0.78 ±0.09, P<0.05).Conclusions Ginsenoside Rg1 could strengthen the spatial learningmemory ability in dementia rats after transplanted with BMSCs.Its mechanism might be possibly correlated with up-regulating mRNA expression of NGF in basal forebrain after BMSCs transplantation

  18. 脐带源间充质干细胞与大鼠胰腺细胞共培养向胰岛样细胞分化及移植治疗1型糖尿病%Islet-like cells derived from mesenchymal stem cells in Wharton's Jelly of the human umbilical cord co-cultured with rat pancreatic cells for transplantation to control type Ⅰ diabetes mellitus

    Institute of Scientific and Technical Information of China (English)

    王广宇; 赵芳; 郝永蕾; 朱旅云; 李晓玲; 胡丽叶; 马利成; 单巍; 杨少玲

    2011-01-01

    BACKGROUND: Mesenchymal stem cells in Wharton's Jelly of the human umbilical cord can induce differentiation into islet-like cells.OBJECTIVE: To verify the possibility of human umbilical cord derived mesenchymal stem cells co-cultured with rat pancreatic cells differentiate into islet-like cells, and to observe the effects of transplantation of islet-like cells on blood glucose of diabetic rats.METHODS: Mesenchymal stem cells in Wharton's Jelly of the human umbilical cord was separated, induced, passaged, and co-cultured with pancreatic cells to induce differentiation into islet-like clusters. Rats were divided into the normal control, model and experimental groups. Rats in the model group were prepared for diabetic models, and those in the experimental group were transplanted islet-like cells after model preparation. RESULTS AND CONCLUSION: There were cells crawled out of cultured Wharton's Jelly of the human umbilical cord, and morphology of adhered cells turned into fusiform shape at 7 days. The isolated cells are characterized by expressing specific surface markers of mesenchymal stem cells, such as CD44, CD29, CD105, but not expressing CD34, CD45 or CD14. The cells were strongly stained by PDX-1 and human insulin at 7 and 10 days. Compared with the simple culture group, the expression of human insulin and concentration of C-peptide were obviously increased; PDX-1 and human insulin mRNA expressions were highly expressed at 7 and 10 days after induction. Compared with the model group, the streptozotocin test of rats in the experimental group was obvious decreased (P < 0.01), but extremely higher than that of the normal control group at 1 week after transplantation (P < 0.01). Brdu positive nuclei and insulin positive kytoplasms could be seen in the experimental group at 8 weeks after transplantation. The results demonstrated that, umbilical cord derived mesenchymal stem cells existed in Wharton's Jelly. The co-cultured cells promote mesenchymal stem cells

  19. PNGF导管复合骨髓间充质干细胞修复大鼠12 mm坐骨神经缺损%PNGF conduit seeded with bone marrow derived mesenchymal stem cells to repair 12-mm sciatic nerve defect of rat

    Institute of Scientific and Technical Information of China (English)

    张伟才; 黄继锋; 严琼娇; 李世普

    2014-01-01

    目的:观察RGD多肽接枝聚(乳酸-羟基乙酸-L-赖氨酸)/聚乳酸/β-磷酸三钙/神经生长因子(PRGD/PDLLA/β-TCP/NGF,PNGF)缓释导管复合骨髓间充质干细胞(Bone marrow derived mesenchymal stem cells, BMSCs)构建组织工程化人工神经,修复大鼠12 mm坐骨神经缺损的效果。方法雄性Wistar大鼠30只,随机分为3组,每组10只,左后肢制作12 mm坐骨神经缺损模型,分别行单纯PNGF导管桥接(A)、PNGF导管复合BMSCs桥接(B)、自体神经移植(C),所有大鼠左侧为实验侧,右侧为正常自身对照侧。术后3个月行大体观察、坐骨神经功能指数、电生理检测、小腿三头肌湿重恢复率测量、新生神经及靶肌肉组织学观察等检测坐骨神经功能恢复情况。结果术后3个月取材时见导管管壁变薄,表面血管化良好,管内有再生神经通过,直径较正常神经细。坐骨神经功能指数的检测结果显示PNGF导管复合BMSCs高于单纯PNGF导管组(P<0.05),PNGF导管复合BMSCs组神经传导速度恢复率、小腿三头肌湿重恢复率、有髓神经纤维数量和直径均优于单纯PNGF导管组(P<0.01),取得与自体神经移植组相似的效果。结论 PNGF缓释导管复合BMSCs桥接修复大鼠坐骨神经缺损,能够有效促进神经再生,效果接近自体神经移植。%Objective To evaluate the effectiveness of tissue-engineered artificial nerve consists of novel arginine-glycine-aspatic acid (RGD) peptide modification of poly{(lactic acid)-co-[(glycolic acid)-alt-(L-lysine)]}/poly(d,l-lactic acid)/β-tricalcium phosphate/nerve growth factor(PRGD/PDLLA/β-TCP/NGF, PNGF) sustaining release conduit seeded with Bone marrow derived mesenchymal stem cells(BMSCs) in bridging 12-mm sciatic nerve defect in rats. Methods 30 isogenic adult male Wister rats were randomly divided into 3 groups (n=10 in each), the left sciatic nerve was exposed to create a 12-mm gap in all

  20. Co-transplantation of bone marrow derived-mesenchymal stem cells and bioactive hydrogel for myocardium infarction therapy in type 2 diabetic rats%骨髓间充质干细胞联合活性水凝胶治疗2型糖尿病大鼠心肌梗死研究

    Institute of Scientific and Technical Information of China (English)

    柯亭羽; 姚欣鹏; 彭嘉睿; 杜娟; 李孔龙; 牛学琴

    2015-01-01

    Objective To evaluate the therapeutic effect of co-transplantation of bone marrow derived-mesenchymal stem cells (BM-MSCs) with Matrigel for treatment of acute myocardial infarction(AMI) in type 2 diabetic(T2DM) rats. Methods T2DM models were induced with 3-week-old SD rats(n=20) with body weight of 50 g, which were randomly divided into 4 groups(n=5):hydrogel co-transplantation group, cell transplantation group, 0.9%sodium chloride solution group and sham operation group. The left anterior descending coronary artery of T2DM SD rats were permanently ligated, and the rats suffered AMI received transplantation of 100μL mixture of Matrigel and 1 × 106 BM-MSCs(hydrogel co-transplantation group), 1 × 106 BM-MSCs suspended in 100μL 0.9%sodium chloride solution(cell transplantation group), and 100μL 0.9%sodium chloride solution(0.9%sodium chloride solution group), with the sham operation group as experimental control. The blood glucose, serum lipid and insulin of rats were measured at 28-day post-operation. All of rats were sacrificed and the heart remodeling was evaluated by histological staining. Immunofluorescent staining was performed to detect the blood vessels reconstruction in MI heart. Results The BM-MSCs isolated from SD rats were typically long fusiform and demonstrated adhesion dependent growth. The level of total cholesterol, low density lipoprotein cholesterol, and insulin in serum of SD rats were obviously up-regulated after 8-week high fat and high fructose diet. After Streptozotocin injection, the fasting blood-glucose of SD rats was all above 7.0 mmol/L, which indicated that the diabetic rat models were successfully constructed. Compared with the saline solution group, transplantation of BM-MSCs obviously improved the heart remodeling post MI, which was favorable for angiogenesis in ischemic heart as determined by immunofluorescent staining. Furthermore, co-transplantation with Matrigel was improved the therapeutic effect of BM-MSCs for MI

  1. Conditioned Medium From Human Amniotic Mesenchymal Stromal Cells Limits Infarct Size and Enhances Angiogenesis

    OpenAIRE

    Danieli, Patrizia; Malpasso, Giuseppe; Ciuffreda, Maria Chiara; Cervio, Elisabetta; Calvillo, Laura; Copes, Francesco; Pisano, Federica; Mura, Manuela; Kleijn, Lennaert; De Boer, Rudolf A.; Viarengo, Gianluca; Rosti, Vittorio; Spinillo, Arsenio; Roccio, Marianna; Gnecchi, Massimiliano

    2015-01-01

    The goal of this study was to elucidate whether human amniotic membrane-derived mesenchymal stromal cells (hAMCs) can exert beneficial paracrine effects on infarcted rat hearts. In particular, the administration of hAMC-conditioned medium repaired ischemic damage through cardioprotection and angiogenesis. Finally, several putative active paracrine mediators that might account for the effects observed were identified by gene and protein arrays.

  2. Mesenchymal stromal cells reset the scatter factor system and cytokine network in experimental kidney transplantation

    OpenAIRE

    Gregorini, Marilena; Bosio, Francesca; Rocca, Chiara; Corradetti, Valeria; Valsania, Teresa; Pattonieri, Eleonora Francesca; Esposito, Pasquale; Bedino, Giulia; Collesi, Chiara; Libetta, Carmelo; Frassoni, Francesco; Canton, Antonio Dal; Rampino, Teresa

    2014-01-01

    Background In former studies we showed in a rat model of renal transplantation that Mesenchymal Stromal Cells (MSC) prevent acute rejection in an independent way of their endowing in the graft. In this study we investigated whether MSC operate by resetting cytokine network and Scatter Factor systems, i.e. Hepatocyte Growth Factor (HGF), Macrophage Stimulating Protein (MSP) and their receptors Met and RON, respectively. Methods MSC were injected into the renal artery soon after reperfusion. Co...

  3. Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells

    OpenAIRE

    Armando de M. Carvalho; Ana Lucia M. Yamada; Juliana R.B. Martins; Leandro Maia; Marjorie de A Golim; Elenice Deffune; Carlos A. Hussni; Ana Liz G. Alves

    2013-01-01

    The objective of the study was to isolate, cultivate and characterize equine peripheral blood-derived multipotent mesenchymal stromal cells (PbMSCs). Peripheral blood was collected, followed by the isolation of mononuclear cells using density gradient reagents, and the cultivation of adherent cells. Monoclonal mouse anti-horse CD13, mouse anti-horse CD44, and mouse anti-rat CD90 antibodies were used for the immunophenotypic characterization of the surface of the PbMSCs. These cells were also ...

  4. Pretreatment of Cardiac Stem Cells With Exosomes Derived From Mesenchymal Stem Cells Enhances Myocardial Repair

    OpenAIRE

    Zhang, Zhiwei; Yang, Junjie; Yan, Weiya; Li, Yangxin; Shen, Zhenya; Asahara, Takayuki

    2016-01-01

    Background Exosomes derived from mesenchymal stem cells (MSCs) were proved to boost cell proliferation and angiogenic potency. We explored whether cardiac stem cells (CSCs) preconditioned with MSC exosomes could survive and function better in a myocardial infarction model. Methods and Results DiI‐labeled exosomes were internalized with CSCs. They stimulated proliferation, migration, and angiotube formation of CSCs in a dose‐dependent manner. In a rat myocardial infarction model, MSC exosome–p...

  5. Pancreatic mesenchyme regulates epithelial organogenesis throughout development.

    Directory of Open Access Journals (Sweden)

    Limor Landsman

    2011-09-01

    Full Text Available The developing pancreatic epithelium gives rise to all endocrine and exocrine cells of the mature organ. During organogenesis, the epithelial cells receive essential signals from the overlying mesenchyme. Previous studies, focusing on ex vivo tissue explants or complete knockout mice, have identified an important role for the mesenchyme in regulating the expansion of progenitor cells in the early pancreas epithelium. However, due to the lack of genetic tools directing expression specifically to the mesenchyme, the potential roles of this supporting tissue in vivo, especially in guiding later stages of pancreas organogenesis, have not been elucidated. We employed transgenic tools and fetal surgical techniques to ablate mesenchyme via Cre-mediated mesenchymal expression of Diphtheria Toxin (DT at the onset of pancreas formation, and at later developmental stages via in utero injection of DT into transgenic mice expressing the Diphtheria Toxin receptor (DTR in this tissue. Our results demonstrate that mesenchymal cells regulate pancreatic growth and branching at both early and late developmental stages by supporting proliferation of precursors and differentiated cells, respectively. Interestingly, while cell differentiation was not affected, the expansion of both the endocrine and exocrine compartments was equally impaired. To further elucidate signals required for mesenchymal cell function, we eliminated β-catenin signaling and determined that it is a critical pathway in regulating mesenchyme survival and growth. Our study presents the first in vivo evidence that the embryonic mesenchyme provides critical signals to the epithelium throughout pancreas organogenesis. The findings are novel and relevant as they indicate a critical role for the mesenchyme during late expansion of endocrine and exocrine compartments. In addition, our results provide a molecular mechanism for mesenchymal expansion and survival by identifying β-catenin signaling as an

  6. The experimental research of peripheral nerve regeneration with rat bone marrow mesenchymal stem cells induced by GD-NF gene%GDNF 基因诱导大鼠骨髓间充质干细胞促周围神经再生的实验研究

    Institute of Scientific and Technical Information of China (English)

    尚修超; 顾加祥; 张乃臣; 刘宏君; 田恒; 潘俊博

    2014-01-01

    Objective To clarify the role of Glial cell derived neurotrophic factor (GDNF)inducing bone marrow mesenchymal stem cells in promoting regeneration of peripheral nerve.Methods Sciatic nerve injury models of rats were established,Experiment was divided into two groups:experimental group was BMSCs group induced by GDNF gene,and control group was BMSCs group un-induced by GDNF gene ,each group had 20 SD rats.① Observe index of sciatic nerve and gastrocnemius wet quality recovery rate after 4 weeks and 8 weeks.② Measured axon diameter ,thick-ness of myelin sheath and myelinated nerve fiber numbers of regeneration nerve fiber 8 weeks postoperative.③The gas-trocnemius muscle cells were stained with DAPI after 4 weeks postoperative.④ Neurofilament regeneration crosslink-ing of the sciatic nerve were obsvered by sweep electron microscope after 8 weeks.Results Test index of the BMSCs group induced by the GDNF gene is better than those of the control group,peripheral nerve regeneration of experimen-tal group observed by scanning electron microscopy was better than those of control group.Conclusion bone marrow mesenchymal stem cells induced by GDNF gene have significant effection for the regeneration of peripheral nerve.%目的:探讨胶质细胞源性神经营养因子(GDNF)诱导骨髓间充质干细胞(BMSCs)促周围神经再生。方法建立大鼠坐骨神经损伤模型,实验分为2组:对照组为未经诱导 BMSCs 组,实验组为 GDNF 基因诱导后 BMSCs组。每组20只 SD 大鼠。①术后4周和8周观察坐骨神经指数和腓肠肌湿质量恢复率。②术后8周测量再生神经纤维轴突直径、髓鞘厚度及有髓神经纤维计数。③术后4周腓肠肌肌细胞行 DAPI 染色。④术后8周坐骨神经扫面电镜观察神经丝再生交联情况。结果经 GDNF 基因诱导 BMSCs 组各方面检测指标均较对照组为好,其中诱导后实验组扫描电镜下观察周围神经再生更加明显。结论GDNF

  7. Enhancement of Bone Marrow Mesenchymal Stem Cells Homing to Kidney Tissue in Acute Renal Failure in Rats by Ultrasound Destruction of Microbubbles%超声辐照微泡促骨髓间充质干细胞归巢于大鼠肾组织实验研究

    Institute of Scientific and Technical Information of China (English)

    唐海林; 王志刚; 李巧; 凌智瑜; 钟世根; 李攀; 李奥

    2012-01-01

    目的 观察超声辐照微泡对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)归巢于急性肾小管坏死模型肾组织的作用.方法 将制备成功的40只肾小管坏死模型大鼠随机分为5组(8只),即(1)单纯模型组(对照组);(2)0.5 W/cm2超声+微泡组(0.5 US+MB);(3)干细胞组(MSCs); (4) 0.5 W/cm2超声+微泡+干细胞组(0.5 US+ MB+MSCs);(5)1.0 W/cm2超声+微泡+干细胞组(1.0 US+ MB+ MSCs).7d后将各组大鼠处死,取材用于荧光显微镜观察MSCs在肾组织的分布情况,采用荧光定量PCR测定细胞间黏附分子1(ICAM-1)基因表达水平.结果 采用荧光显微镜观察,DAPI标记的MSCs细胞核(细胞核带蓝色荧光),1.0 US+ MB+ MSCs组肾组织骨髓干细胞数量明显多于0.5 US+ MB+ MSCs组及MSCs组(P<0.05).0.5 US+MB组、0.5 US+ MB+MSCs组及1.0 US+ MB+MSCs组ICAM-1的基因水平明显高于对照组及MSCs组(P<0.05).结论 1.0 W/cm2超声辐照微泡促进肾组织细胞ICAM-1表达增加,从而促进MSCs归巢于急性肾小管坏死模型的肾组织.%Objective To explore the effect of bone marrow mesenchymal stem cells homing to kidney tissue in acute renal failure in rats by ultrasound destruction of microbubbles. Methods Acute renal failure (ARF) was induced by subcutaneous injection of HgCl2. Forty Sprague-Dawley (SD) rats were randomly divided into the following group after the establishment of animal models of acute renal failure (n = 8). (1) Model group alone (control group); (2) 0. 5 W/cm2 US+MB group, Bubble (0. 5 ml)+0. 5 W/cm2 US; (3) MSCs group; (4) 0. 5 W/cm2 US+MB+MSCs group; (5) 1.0 W/cm2 US+MB+MSCs group. MSCs were Labelled with DAPI. The number of DAPI-positive MSCs was evaluated by fluorescence microscopy, and real-time PCR was performed 7 days after MSC transplantation. Results It was observed via fluorescence microscopy that the number of DAPI-positive MSCs in the kidney for the 1. 0 W/cm2 US+MB+MSCs group was significantly more than that of

  8. Chemopreventive effect of Quercus infectoria against chemically induced renal toxicity and carcinogenesis

    Directory of Open Access Journals (Sweden)

    Muneeb U Rehman

    2012-06-01

    Full Text Available In this study we have shown that Quercus infectoria attenuates Fe- NTA induced renal oxidative stress, hyperproliferative response and renal carcinogenesis in rats. Fe-NTA promoted DEN (N-diethyl nitrosamine initiated renal carcinogenesis by increasing the percentage incidence of tumors and induces early tumor markers viz. ornithine decarboxylase (ODC level and PCNA expression. Fe- NTA (9 mg Fe/kg body weight, intraperitoneally enhances renal Malondialdehyde, xanthine oxidase and hydrogen peroxide generation with reduction in renal glutathione content, antioxidant enzymes, viz., glutathione peroxidase, glutathione reductase, catalase, glucose-6-phosphate dehydrogenase and phase-II metabolizing enzymes such as glutathione-S-transferase and quinone reductase. It also enhances blood urea nitrogen and serum creatinine. Fe-NTA also lead to increase in levels of some inflammatory markers viz NO and MPO and some proinflammatory cytokines viz PGE-2 and TNF-1. The chemopreventive efficacy of Quercus infectoria was studied in terms of xenobiotic metabolizing enzyme activities, LPO, redox status, serum toxicity markers, inflammatory and proinflammatory markers and cell proliferation in the kidney tissue. Oral administration of Quercus infectoria at doses of 75 and 150 mg/kg b wt effectively suppressed renal oxidative stress, inflammation and tumor incidence. Chemopreventive effects of Quercus infectoria were associated with up-regulation of xenobiotic metabolizing enzyme activities and down regulation of serum toxicity markers. Present study supports Quercus infectoria as a potent chemopreventive agent and suppresses Fe-NTA-induced renal carcinogenesis and oxidative and inflammatory response in Wistar rat.

  9. Prevention of chemically induced diabetes mellitus in experimental animals by virgin argan oil.

    Science.gov (United States)

    Bellahcen, Said; Mekhfi, Hassane; Ziyyat, Abderrahim; Legssyer, Abdelkhaleq; Hakkou, Abdelkader; Aziz, Mohammed; Bnouham, Mohamed

    2012-02-01

    The argan tree plays an important socioeconomic and ecologic role in South Morocco. Moreover, there is much evidence for the beneficial effects of virgin argan oil (VAO) on human health. Thus, this study investigated whether administering VAO to rats can prevent the development of diabetes. VAO extracted by a traditional method from the almonds of Argania spinosa (2 mL/kg) was administered orally (for 7 consecutive days) to rats before and during intraperitoneal alloxan administration (75 mg/kg for 5 consecutive days). An alloxan diabetic-induced untreated group and treated by table oil were used as control groups. Body mass, blood glucose and hepatic glycogen were evaluated. In the present study, subchronic treatment with VAO at a dose of 2 mL/kg, before the experimental induction of diabetes, prevented the body mass loss, induced a significant reduction of blood glucose and a significant increase of hepatic glycogen level (p argan oil should be further investigated in a human study to clarify its possible role in reducing weight loss in diabetics, and even in inhibiting the development or progression of diabetes. This antidiabetic effect could be due to the richness of VAO in tocopherols, phenolic compounds and unsaturated fatty acids. PMID:21584872

  10. 鼠骨髓间充质干细胞对缺氧/复氧心肌细胞的影响%Effect of bone marrow mesenchymal stem cells on hypoxia/reoxygenation cardiac myocytes in rats

    Institute of Scientific and Technical Information of China (English)

    章培军; 张丽红; 郭敏芳

    2010-01-01

    目的:观察在体外培养条件下骨髓间充质干细胞(mesenchymal stem cell,MSC)对缺氧/复氧新生大鼠心肌细胞的保护作用.方法:无菌条件下分离、培养、传代Wistar大鼠MSC,细胞融合达90%时更换培养基培养24 h,收集细胞培养液作为MSC条件培养基;原代培养新生Wistar大鼠心肌细胞,先用5% CO2、10% H2、85% N2混合气造成细胞缺氧24 h,再给予95% O2 和 5% CO2混合气复氧2 h,建立缺氧/复氧模型.MSC 处理组于复氧时加入MSC 条件培养基,正常细胞作为对照组,分别于复氧后用MTT法、乳酸脱氢酶(lactated dehydrogenase)法测定各组心肌细胞活性的改变;RT-PCR观察细胞凋亡蛋白Bcl-2和Bax的表达.结果: 与对照组比较,缺氧/复氧组细胞活力显著降低(P < 0.01).与缺氧/复氧组比较,MSC处理组细胞活力明显增加(P < 0.01).缺氧/复氧组抗凋亡基因Bcl-2的表达明显降低,促凋亡基因Bax的表达明显增加,而MSC条件培养基可以减少Bax基因的表达,提高Bcl-2/Bax值,与缺氧/复氧组比较差异具有显著性(P < 0.01).结论:MSC对体外缺氧-复氧诱导的心肌细胞的凋亡有保护作用.

  11. Influence of different low oxygen partial pressures on proliferation and differentiation of mesenchymal stem cells in SD rats%不同低氧分压对SD大鼠骨髓间充质干细胞增殖分化的影响

    Institute of Scientific and Technical Information of China (English)

    彭涛; 黄姣; 徐凌

    2012-01-01

    Objective To investigate the effects of different low partial pressure of oxygen on proliferation and differentiation capability of rat mesenchymal stem cells(MSCs) in vitro. Methods MSCs were isolated from adult rat bone marrow and cultured in vitro under normoxia(21% oxygen tension) and low PO2 (2% ,4% ,6% and 8% oxygen tension) respectively. The proliferation of MSCs, alkaline phosphatase(ALP) activity,and production of collagens type Ⅰ (COLⅠ ) were examined by means of MTT and enzyme-linked immunoreaction onl,3,5,7,9 d respectively. Results Different low partial oxygen pressures had significant promoting effect on the proliferation activity of MSCs(P<0. 05) and decreased ALP activity and the production of COL Ⅰ (P<0. 05). Conclusion Hypoxia microenvironment can promote the proliferation activity of MSCs and inhibit MSCs to differentiate into osteogenic lineage cells.%目的 研究体外不同低氧分压对SD大鼠骨髓间充质干细胞(MSCs)增殖分化的影响.方法 将体外培养的MSCs,分组置于常氧(21%)和低氧(2%、4%、6%、8%)培养箱中培养,用MTT法分别于1、3、5、7、9 d检测细胞的增殖活性;用酶联免疫检测法分别于1、3、5、7、9 d检测细胞碱性磷酸酶(ALP)和Ⅰ型胶原蛋白(COLⅠ)的表达.结果 不同低氧分压对MSCs细胞增殖活性有明显促进作用(P<0.05);低氧分压抑制MSCs 细胞的ALP和COLⅠ表达(P<0.05).结论 低氧微环境可促进MSCs的增殖活性,但对细胞的成骨向分化起到抑制作用.

  12. 同种异体骨髓间充质干细胞移植治疗糖尿病大鼠的实验研究%An experimental study on treatment of diabetic rats through transplantation of allogeneic bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    高斌; 宓真; 杜馨丽; 车海龙

    2011-01-01

    Objective: To observe the curative effectiveness of allogeneic bone marrow mesenchymal stem cells (BMSC) transplantation in diabetic rats and the possible mechanism.Methods: BMSC were isolated from allogeneic bone marrow of SD rat by adhesive screening method and cultured in vitro.The second passage of BMSC was transfected with Ad-GFP.BMSC that stably expressed CFP were obtained before transplantation.Streptozotocin-induced diabetic rats (plasma glucose > 16.7 mmol/L) were divided at random into untreated group (UTG) and BMSC transplantation group (TPG).BMSC were transplanted into allogeneic diabetic rats of TPG through vena caudalis (0.4 ml, 1×106-1×l07/ml).Another 5 normal rats picked out at random (normal control group) and the diabetic rats of UTG were injected via tail vein with the same quantity of physiological saline.After the 2 weeks transplantation, the levels of plasma glucose, HbA1c and insulin were determined.The pancreas was dissected out for pathological sections.Insulin reaction with immunohistochemistry of rats was observed.Fluorescent microscopy was used to observe the expression of GFP labeled cells in the pancreas.Results: The result showed that levels of plasma glucose and HbA1c were higher obviously in UTG than in normal control group (NCG), and insulin level lower (P<0.001).The levels of plasma glucose and HbA,c were lower obviously in TPG than in UTG (P<0.05), and insulin level higher (P>0.05).The numbers of beta-cells of the UTG decreased and the insulin reaction showed negative results.The numbers of beta-cells of the TPG increased and the insulin reaction showed positive and strong positive results.The cells of GFP were observed in the excrine tissues of pancreas, but not observed in the islet.Conclusion: It is concluded that BMSC transplantation by tail vein injection could engraft into pancreas tissues of diabetic rats, depress the levels of plasma glucose and HbA1c, accelerate the synthesis of the insulin.BMSC transplantation

  13. 骨髓间充质干细胞预处理脑死亡供鼠对移植肾的保护作用%The protective effects on the renal allografts from brain dead donor rats pretreated with bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    陈洁; 张智; 曾慧兰; 苏泽轩; 余钧雷; 吴永璐; 袁博翔

    2015-01-01

    目的 探讨骨髓间充质干细胞(MSC)预处理脑死亡(BD)供鼠对移植肾的保护作用.方法 实验分正常移植组、脑死亡供肾移植组(BD移植组)和输注MSC+脑死亡供肾移植组(MSC处理组),雄性F344大鼠为供者,雄性Lewis大鼠为受者,正常移植组为常规移植;BD移植组为脑死亡F344大鼠供肾;MSC处理组在移植前输注预先制备的来自F344大鼠的MSC,然后接受脑死亡F344大鼠供肾移植.移植后给予环孢素A,连用10天.术后第10天切除其右肾.测定术后第14、21、28和35天血肌酐水平.术后35 d获取移植肾标本,行病理学观察;免疫组化检测移植肾组织中白细胞介素(IL)-1β和肿瘤坏死因子α(TNF-α)表达情况.结果 BD移植组术后各检测时点的血肌酐水平均高于其余两组(P<0.05),而MSC处理组和正常移植组间除第21天的血清肌酐值外,其余时间段的差异没有统计学意义.BD移植组移植肾组织中可见单核细胞浸润,其肾小管上皮炎症明显重于其他两组,MSC处理组与正常移植组的差异不大.BD移植组移植肾肾小球上皮细胞、肾小管上皮细胞、间质细胞的IL-1β和TNF-α的表达均呈阳性,染色程度较强,明显强于其他两组(H=7.210,P=0.027),而MSC处理组和正常移植组间的差异无统计学意义.结论MSC预处理脑死亡供鼠,可能通过减少移植肾炎症细胞浸润,降低TNF-α和IL-1β表达来减轻移植肾损伤.%Objective To investigate the protective effects on the renal allografts from brain dead (BD) donor rats pretreated with bone marrow mesenchymal stem cells (MSCs).Method Three groups [normal transplant group (G1).BD transplant group (G2),and MSCs pretreated + BD transplant group (G3)] were set up.Male F344 rats served as donors and male Lewis rats as recipients.In G1,kidneys from F344 donor rats were implanted into Lewis recipients.In G2,kidneys from F344 BD donor rats were engrafted into Lewis recipients.In G3,after BD was

  14. Combination of high-performance refractometry and infrared spectroscopy as a probe for chemically induced gelation and vitrification of epoxies

    Energy Technology Data Exchange (ETDEWEB)

    Mueller, U; Philipp, M; Gervais, P-C; Sanctuary, R; Krueger, J K [Laboratoire de Physique des Materiaux, Universite du Luxembourg, 162A avenue de la faiencerie, L-1511 Luxembourg (Luxembourg); Possart, W; Wehlack, C [Fachbereich Werkstoffwissenschaften, Universitaet des Saarlandes, D-66123 Saarbruecken (Germany); Kieffer, J, E-mail: ulrich.mueller@uni.l [Department of Materials Science and Engineering, University of Michigan, Ann Arbor, MI (United States)

    2010-08-15

    A combination of infrared spectroscopy and high-performance refractometry was used to investigate the chemically induced sol-gel and glass transition during the polymerization of epoxies. Representations of the refractive index versus chemical conversion reveal an interesting insight into the optical properties accompanying gelation and vitrification. Whereas the electronic polarizability of the liquid state of small average molecular mass and the glassy state is dominated by the mass density, an unexpected excess polarizability observed during the gelation is attributed to cooperative dipole-dipole interactions.

  15. Chemically induced dynamic electron polarization investigation of the triplet-radical system in the solution of the triplet quencher

    Institute of Scientific and Technical Information of China (English)

    LU, Tong-Xing; CUI, Zhi-Feng; XU, Xin-Sheng; ZHANG, Xian-Yi

    2000-01-01

    The chemically induced dynamic electron polariztiion (CIDEP) of the triplet molecule/triplet quencher/2,2,6,6-te tranethyl-1-piperidinyioxyl (TEMPO) systems were measured using the high time-resolved FESR spectrometer. The competi tion between the radical-triplet pair mechanism (RTPM) and triplet mechanism (TM) or radical pair mechanism (RPM) polarization in the solution of the triplet quencher was investi gated, and the relationship between reaction rate of the radi cal-triplet pair and quenching rate of triplet was deduced.

  16. 通过磁共振来评价大鼠和兔子海绵体中人间叶细胞干细胞的分布%Magnetic resonance evaluation of human mesenchymal stem cells in corpus cavernosa of rats and rabbits

    Institute of Scientific and Technical Information of China (English)

    Yun-Seob Song; Dongho Choi; Jong-Ho Won; Ja-Hyeon Ku; Eun-Seop Song; Jung-Hoon Kim; Jin-Suck Jeon; Kong-Hee Lee; Sook-Ja Kim; Hee-Jeong Cheong; Ik-Sung Lim

    2007-01-01

    Aim: To investigate whether the biological process of superparamagnetic iron oxide (SPIO)-labeled human mesenchymal stem cells (hMSCs) may be monitored non-invasively by using in vivo magnetic resonance (MR) imaging with conventional 1.5-T system examinations in corpus cavernosa of rats and rabbits. Methods: The labeling efficiency and viability of SPIO-labeled hMSCs were examined with Prussian blue and Tripan blue, respectively. After SPIO-labeled hMSCs were transplanted to the corpus cavernosa of rats and rabbits, serial T2-weighted MR images were taken and histological examinations were carried out over a 4-week period. Results: hMSCs loaded with SPIO compared to unlabeled cells had a similar viability. For SPIO-labeled hMSCs more than 1×105 concentration in vitro,MR images showed a decrease in signal intensity. MR signal intensity at the areas of SPIO-labeled hMSCs in the rat and rabbit corpus cavernosa decreased and was confined locally. After injection of SPIO-labeled hMSCs into the corpus cavernosum, MR imaging demonstrated that hMSCs could be seen for at least 12 weeks after injection. The presence of iron was confirmed with Prussian blue staining in histological sections. Conclusion: SPIO-labeled hMSCs in corpus cavernosa of rats and rabbits can be evaluated non-invasively by molecular MR imaging. Our findings suggest that MR imaging has the ability to test the long-term therapeutic potential of hMSCs in animals in the setting of erectile dysfunction.%目的:研究能否用常规的1.5-T体内磁共振成像检测系统非侵入性地监控超顺磁性氧化铁(SPIO)标记的人间叶细胞干细胞(hMSCs)在大鼠和兔子的海绵体内的生物学过程.方法:分别用普鲁士蓝和台盼蓝法检测hMSCs的SPIO标记效率和其活力.将SPIO标记的hMSCs移植到大鼠和兔子的海绵体后,用系列T2磁共振成像进行组织学检测,历时4周.结果:SPIO标记的hMSCs与没有标记的细胞有相似的活力.当SPIO标记的h

  17. Human stromal (mesenchymal) stem cells

    DEFF Research Database (Denmark)

    Aldahmash, Abdullah; Zaher, Walid; Al-Nbaheen, May;

    2012-01-01

    Human stromal (mesenchymal) stem cells (hMSC) represent a group of non-hematopoietic stem cells present in the bone marrow stroma and the stroma of other organs including subcutaneous adipose tissue, placenta, and muscles. They exhibit the characteristics of somatic stem cells of self......-renewal and multi-lineage differentiation into mesoderm-type of cells, e.g., to osteoblasts, adipocytes, chondrocytes and possibly other cell types including hepatocytes and astrocytes. Due to their ease of culture and multipotentiality, hMSC are increasingly employed as a source for cells suitable for a number...

  18. Behavioral changes in rats with focal cerebral ischemia after directional transplantation of bone marrow mesenchymal stem cells%骨髓间充质干细胞定向移植局灶性脑缺血大鼠的行为学变化

    Institute of Scientific and Technical Information of China (English)

    马学玲; 王心蕊; 江新梅; 陈柏竹; 李霞; 刘亢丁

    2008-01-01

    BACKGROUND: Transplantation of bone marrow mesenchymal stem cells (MSCs) has been used in the field of repair of nerve injury. Brain stereotactic transplantation and transvascular transplantation are two transplantation methods. OBJECTIVE: We infused MSCs into rat peripheral cerebral infarct focus, in order to investigate the improvement of rat neurological dysfunction by forelimb use asymmetry test and postural reflex test.DESIGN: A randomized controlled animal experiment. SETTING: Department of Neurology, First Hospital of Jilin University.MATERIALS: This study was performed at the Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, Jilin University between October 2006 and April 2007. Healthy male Wistar rats of clean grade, weighing 250-280 g, were provided by the Laboratory Animal Center of Jilin University. The protocol was performed in accordance with ethical guidelines for the use and care of animals.METHODS: MSCs from healthy adult volunteers were in vitro cultured and proliferated by density gradient separation and adherence screening method. Their immunophenotypes were identified by a flow cytometer. The Wistar rats were randomized into 5 groups with 10 rats in each: normal control group, sham-operated group, model group, serum-free DMEM-treated group (DMEM group) and MSCs -treated group (MSCs group). Rat models of cerebral ischemia/reperfusion were developed by occluding rat right middle cerebral artery following suture occlusion method modified by Longa et al. Rats in the normal control group were untouched. In the sham-operated group, operation was not ended till cervical interior and exterior arteries were exposed and sutured, and the other disposals were the same as those in the model group. At ischemia 90 minutes reperfusion 1 hour, a stereotaxic apparatus was used to take rat right peripheral cerebral ischemic region as transplantation site: 3 mm lateral to, 1mm caudal to and 4 mm posterior to Bregma. Rats in the MSCs group

  19. Propofol with bone marrow mesenchymal stem cell transplantation improves the hind limb function and electrophysiological changes in rats with spinal cord injury%丙泊酚联合骨髓间充质干细胞移植对脊髓损伤大鼠后肢功能及电生理的影响

    Institute of Scientific and Technical Information of China (English)

    何珏; 王天科

    2015-01-01

    < combination group, and the differences were statisticaly significant (P < 0.05). These findings indicate that both propofol and bone marrow mesenchymal stem cels can promote synaptic regeneration and improve the electrophysiological function and motor function of rats with spinal cord injury. Their combination has a better role than propofol and bone marrow mesenchymal stem cels used alone.

  20. 尾静脉移植骨髓间充质干细胞对酒精性痴呆大鼠的作用%Therapeutic effect of intravenous injection of bone marrow mesenchymal stem cells on alcohol-associated dementia rat

    Institute of Scientific and Technical Information of China (English)

    吴晓牧; 曾玉娥; 刘勇; 何丹; 杨海玉

    2013-01-01

    目的 探讨静脉移植骨髓间充质干细胞(BMMSC)治疗酒精性痴呆模型鼠(AAD)的疗效及其机制.方法 采用全骨髓贴壁法分离培养大鼠BMMSC并进行流式细胞鉴定.通过持续28d酒精灌胃(20%,8ml/kg)建立AAD大鼠模型,经尾静脉途径将DAPI标记的BMMSC移植至大鼠体内,观察其在海马中的定位,并以Morris水迷宫行为学、海马组织形态和神经细胞凋亡检测作为疗效评价标准.采用免疫组化检测大鼠海马区脑源性神经营养因子(BDNF)阳性细胞的表达以及比色法检测大鼠血清总超氧化物歧化酶(T-SOD)和谷胱甘肽过氧化物酶(GSH-Px)的变化.结果 BMMSC高表达CD29、CD90,低表达CD34、CD45,移植后可迁移至大鼠海马区.与PBS组相比,BMMSC移植后大鼠的逃避潜伏期时间明显缩短[(10.17±0.71)s,(4.71±0.34)s,P<0.01],且其海马形态结构破坏及神经细胞凋亡明显改善[(72.67±2.73),(55.5±5.14),P<0.05].免疫组化结果显示,与PBS组相比,BMMSC组大鼠海马区BDNF的表达显著增高[(71.54±13.71),(135.25±22.20),P<0.05].氧化应激检测证实,与PBS组相比,BMMSC组大鼠血清GSH-Px酶活力显著增高[(526.89 ±62.73)vs.(2592.75±243.73),P<0.01],但T-SOD酶活力无明显改变.结论 BMMSC移植可改善AAD大鼠学习记忆功能及修复海马组织损伤,其机制可能与促进海马组织BDNF表达及抗氧化应激有关.%Objective To observe therapeutic benefits of intravenously transplanted hone marrow mesenchymal stem cells (BMMSCs) on alcohol-associated dementia (AAD) rat model and study its underlying mechanisms.Methods BMMSCs were isolated by the method of differential adhesion and membrane antigens were detected with flowcytometric analysis.To establish AAD model,SD rats were intragastricly administrated with ethanol (20%,8ml/kg) for 28 days.Then BMMSCs were labeled with DAPI and injected into the blood via caudal vein.And animals were evaluated by observing Morris Maze behavior

  1. Vascularization of the dorsal root ganglia and peripheral nerve of the mouse: Implications for chemical-induced peripheral sensory neuropathies

    Directory of Open Access Journals (Sweden)

    Melemedjian Ohannes K

    2008-03-01

    Full Text Available Abstract Although a variety of industrial chemicals, as well as several chemotherapeutic agents used to treat cancer or HIV, preferentially induce a peripheral sensory neuropathy what remains unclear is why these agents induce a sensory vs. a motor or mixed neuropathy. Previous studies have shown that the endothelial cells that vascularize the dorsal root ganglion (DRG, which houses the primary afferent sensory neurons, are unique in that they have large fenestrations and are permeable to a variety of low and high molecular weight agents. In the present report we used whole-mount preparations, immunohistochemistry, and confocal laser scanning microscopy to show that the cell body-rich area of the L4 mouse DRG has a 7 fold higher density of CD31+ capillaries than cell fiber rich area of the DRG or the distal or proximal aspect of the sciatic nerve. This dense vascularization, coupled with the high permeability of these capillaries, may synergistically contribute, and in part explain, why many potentially neurotoxic agents preferentially accumulate and injure cells within the DRG. Currently, cancer survivors and HIV patients constitute the largest and most rapidly expanding groups that have chemically induced peripheral sensory neuropathy. Understanding the unique aspects of the vascularization of the DRG and closing the endothelial fenestrations of the rich vascular bed of capillaries that vascularize the DRG before intravenous administration of anti-neoplastic or anti-HIV therapies, may offer a mechanism based approach to attenuate these chemically induced peripheral neuropathies in these patients.

  2. 携带肝细胞生长因子基因的骨髓间充质干细胞移植对肺动脉高压大鼠治疗作用初探%Role of bone marrow mesenchymal stem cells transplantation in treatment of pulmonary arterial hypertension in rats carrying hepatocyte growth factor

    Institute of Scientific and Technical Information of China (English)

    郭娜; 郭英华; 苏龙翔; 常德; 刘岩; 姜学革; 刘长庭

    2012-01-01

    Objective To study the role of bone marrow mesenchymal stem cells(MSCs) transplantation in treatment of monocrotaline (MCT)-induced pulmonary arterial hypertension(PAH) in rats carrying hepatocyte growth factor(HGF) and its mechanism. Methods MSC isolated from bone marrow of 3-week old male SD rats were incubated in vitro with adhesion culture method, purified and transfected with adenovirus(Ad)-mediated HGF. The rats were randomly divided into PAH group, MSC transplantation group, and Ad-mediated HGF transplantation group, 10 in each group. After a PAH model was established by intraperitoneal injection of monocrotaline(MCT) at the dose of 60mg/kg, lml L-DMEMwas injected into the jugular vein of rats in PAH group, lml suspension containing 5 x 106 MSC transfected with empty adenovirus vector was injected into rats of HGF transplantation group. Hemodynamics, RV/LV and plasma ET-1 level in rats of different groups were observed. Results No significant difference was found in BP among different groups 3 weeks after treatment of PAH(pulmonary arterial hypertension). The PASP(pulmonary artery systdic pressure) and mPAP(mean pulmonary arterial pressure) were significantly lower in Ad-mediated HGF transplantation group than in PAH group and MSCs transplantation group(37.227 2 ± 5.381 60 vs 45.069 0 ± 7.177 97 and 44.142 0 ± 6.673 75, 25.693 5 ± 1.446 33 vs 30.800 2 ± 1.256 15 and 30.359 1 ± 1.172 52, P0.05)although it was decreased. The plasma ET-1 level was significantly lower in Ad-mediated HGF transplantation group than in PAH group and MSC transplantation group(23.862 70 ±7.426 76 vs 28.372 0± 4.402 75 and 31.849 0± 7.547 13, P<0.05). Conclusion MSC carrying HGF transplantation therapy can significantly reduce the PASP and mPAP, improve hemodynamics, delay and even reverse right ventricular hypertrophy, and decrease plasma ET-1 level in rats with PAH, thus protecting vascular endothelium cells against damage and blood vessels against constriction.%目的 研

  3. Central nervous system mesenchymal chondrosarcoma

    International Nuclear Information System (INIS)

    Central nervous system mesenchymal chondrosarcomas are rare malignant tumors that constitute a separate entity from the classical chondrosarcoma and myxoid variant. Clinical behaviour of central nervous system chondrosarcomas is still unknown. We describe two rare examples of intracranial mesenchymal chondrosarcoma with a review of the literature, in an attempt to clarify the clinical characteristics, prognosis and treatment of choice of these unusual tumors. Among the 55 reported cases, 23 had postoperative radiotherapy. Although there is no statistical significance according to the Log-Rank test (p=0.7), the patients treated with radiation therapy seem to have a better chance of survival. Patients who had adjuvant chemotherapy (only 5) showed survival times similar to those patients who had none. Although clinical behaviour of central nervous system chondrosarcomas remains to be defined, data from our series as well as literature show that radical removal is the best therapeutic choice. In addition, patients treated with postoperative radiotherapy seem to show a trend toward increased survival

  4. Promoting Nerve Regeneration in a Neurotmesis Rat Model Using Poly(DL-lactide-ε-caprolactone Membranes and Mesenchymal Stem Cells from the Wharton’s Jelly: In Vitro and In Vivo Analysis

    Directory of Open Access Journals (Sweden)

    T. Pereira

    2014-01-01

    Full Text Available In peripheral nerves MSCs can modulate Wallerian degeneration and the overall regenerative response by acting through paracrine mechanisms directly on regenerating axons or upon the nerve-supporting Schwann cells. In the present study, the effect of human MSCs from Wharton’s jelly (HMSCs, differentiated into neuroglial-like cells associated to poly (DL-lactide-ε-caprolactone membrane, on nerve regeneration, was evaluated in the neurotmesis injury rat sciatic nerve model. Results in vitro showed successful differentiation of HMSCs into neuroglial-like cells, characterized by expression of specific neuroglial markers confirmed by immunocytochemistry and by RT-PCR and qPCR targeting specific genes expressed. In vivo testing evaluated during the healing period of 20 weeks, showed no evident positive effect of HMSCs or neuroglial-like cell enrichment at the sciatic nerve repair site on most of the functional and nerve morphometric predictors of nerve regeneration although the nociception function was almost normal. EPT on the other hand, recovered significantly better after HMSCs enriched membrane employment, to values of residual functional impairment compared to other treated groups. When the neurotmesis injury can be surgically reconstructed with an end-to-end suture or by grafting, the addition of a PLC membrane associated with HMSCs seems to bring significant advantage, especially concerning the motor function recovery.

  5. Differentiation of Bone Marrow Mesenchymal Cells to Neural Cells

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    To explore the possibility and condition of differentiation of bone marrow mesenchymal cells (BMSCs) to neural cells in vitro, BMSCs from whole bone marrow of rats were cultured. The BMSCs of passage 3 were identified with immunocytochemical staining of CD44 ( + ), CD71 ( + )and CD45(-). There were type Ⅰ and type Ⅱ cells in BMSCs. Type Ⅰ BMSCs were spindleshaped and strong positive in immunocytochemical staining of CD44 and CD71, whereas flat and big type Ⅱ BMSCs were lightly stained. The BMSCs of same passage were induced to differentiate into neural cells by β-mercaptoethanol (BME). After induction by BME, the type Ⅰ BMSCs withdrew to form neuron-like round soma and axon-like and dendrite-like processes, and were stained positively for neurofilament (NF). The type Ⅱ BMSCs did not change in the BME medium and were negatively or slightly stained of NF.

  6. Migration capacity of human umbilical cord mesenchymal stem cells towards glioma in vivo*

    Institute of Scientific and Technical Information of China (English)

    Cungang Fan; Dongliang Wang; Qingjun Zhang; Jingru Zhou

    2013-01-01

    High-grade glioma is the most common malignant primary brain tumor in adults. The poor prognosis of glioma, combined with a resistance to currently available treatments, necessitates the ment of more effective tumor-selective therapies. Stem cel-based therapies are emerging as novel cel-based delivery vehicle for therapeutic agents. In the present study, we successful y isolated human umbilical cord mesenchymal stem cel s by explant culture. The human umbilical cord senchymal stem cel s were adherent to plastic surfaces, expressed specific surface phenotypes of mesenchymal stem cel s as demonstrated by flow cytometry, and possessed multi-differentiation potentials in permissive induction media in vitro. Furthermore, human umbilical cord mesenchymal stem cel s demonstrated excel ent glioma-specific targeting capacity in established rat glioma models after intratumoral injection or contralateral ventricular administration in vivo. The excellent glioma-specific targeting ability and extensive intratumoral distribution of human umbilical cord mesenchymal stem cel s indicate that they may serve as a novel cel ular vehicle for delivering the-rapeutic molecules in glioma therapy.

  7. Protection of tetracycline on damage of rat bone mesenchymal stem cells induced by glucocorticosteroid%四环素对糖皮质激素诱导的骨髓间充质干细胞损伤的保护作用

    Institute of Scientific and Technical Information of China (English)

    杨智洋; 张德志; 仲兴; 崔凤荣; 郭涛; 张男; 刘丹平

    2011-01-01

    Objective To investigate the effect of tetracycline on rat bone mesenchymal stem cells (BMSCs) injury indued by glucocorticosteroid. Methods The model of rat BMSCs injury induced by glucocorticosteroid in vitro was estab lished, and then isolately cultured in vitro. The BMSCs of rats were divided into six groups; normal control group, hormone control group, 4, 20, 100, 500 u,g/ml tetracycline group. The injury of BMSCs induced by the glucocorticosteroid was de tected by Acridine orange-ethidium bromide double staining. The cytotoxicity of BMSCs was measured by MTT assay, and then drawing cell growth curve. Staining and analysis of bcl-2, bax epressions were detected by immunohistochemical. Western blot was used to detect the expression of caspase-3. Results Acridine orange-ethidium bromide double staining showed that the cell damage was apoptosis. Tetracycline had protection effect on cell proliferation of BMSCs inhibited by glucocorticosteroid by dose-dependent mode. 100 μg/ml tetracycline could induce the apoptosis of BMSCs by increasing the bcl-2, decreasing bax、caspas-3. Conclusion Tetracycline can protect the injury of BMSCs induced by glucocorticoste roid. This provide some experimental pharmaco-foundation for intervention therapy of early osteonecrosis.%目的 探讨四环素对糖皮质激素诱导大鼠骨髓间充质干细胞(BMSCs)损伤的保护作用.方法 体外建立糖皮质激素诱导大鼠BMSCs损伤模型,分离与体外培养大鼠BMSCs.将大鼠BMSCs分为正常对照组、激素对照组,四环素4、20、100、500 μg/ml浓度组.采用吖啶橙/溴化乙锭双荧光染色法观察四环素对激素诱导的BMSCs凋亡的影响;采用MTT法检测各组BMSCs活性,绘制细胞生长曲线;用免疫组织化学方法对细胞凋亡因子bcl-2、bax进行分析;Western blot测定细胞凋亡因子caspase-3表达.结果 吖啶橙/溴化乙锭染色显示细胞损伤为凋亡.四环素对激素抑制BMSCs的增殖有明显保护作用,

  8. Aging impairs osteoblast differentiation of mesenchymal stem cells grown on titanium by favoring adipogenesis

    Science.gov (United States)

    ABUNA, Rodrigo Paolo Flores; STRINGHETTA-GARCIA, Camila Tami; FIORI, Leonardo Pimentel; DORNELLES, Rita Cassia Menegati; ROSA, Adalberto Luiz; BELOTI, Marcio Mateus

    2016-01-01

    ABSTRACT Aging negatively affects bone/titanium implant interactions. Our hypothesis is that the unbalance between osteogenesis and adipogenesis induced by aging may be involved in this phenomenon. Objective We investigated the osteoblast and adipocyte differentiation of mesenchymal stem cells (MSCs) from young and aged rats cultured on Ti. Material and Methods Bone marrow MSCs derived from 1-month and 21-month rats were cultured on Ti discs under osteogenic conditions for periods of up to 21 days and osteoblast and adipocyte markers were evaluated. Results Cell proliferation, alkaline phosphatase (ALP) activity, extracellular matrix mineralization and gene expression of RUNX2, osterix, ALP, bone sialoprotein, osteopontin, and osteocalcin were reduced in cultures of 21-month rats compared with 1-month rats grown on Ti. Gene expression of PPAR-γ , adipocyte protein 2, and resistin and lipid accumulation were increased in cultures of 21-month rats compared with 1-month rats grown on the same conditions. Conclusions These results indicate that the lower osteogenic potential of MSCs derived from aged rats compared with young rats goes along with the higher adipogenic potential in cultures grown on Ti surface. This unbalance between osteoblast and adipocyte differentiation should be considered in dental implant therapy to the elderly population. PMID:27556209

  9. Generation of lymphomagenic mouse type-C viruses from radiation- or chemically-induced non-producer T-cell lymphoma cell lines infected with non-oncogenic ecotropic virus

    International Nuclear Information System (INIS)

    Highly lymphomagenic mouse type-C viruses were generated from radiation- or chemically-induced T-cell lymphoma cell lines of NFS/N mouse origin infected with a non-oncogenic ecotropic virus E4. By analysis of these progeny viruses, the following results were obtained. 1) The viruses were lymphomagenic in neonatally inoculated NFS/N and C3H/He mice and W/Fu rats but not in Balb/c and C57BL/6N mice, indicating that they possess the Fv-1n tropism of exogenously infected parent virus. 2) Lymphomagenic viruses consisted of plural viral subpopulations. Recombinant mink cell focus-inducing (MCF) and ecotropic viruses were cloned from them. Inoculation of either MCF or ecotropic virus alone or both viruses together did not cause lymphoma in NFS/N mice and there was no evidence of viral replication in the recipients. 3) Inoculation of either MCF- or ecotropic virus-infected NFS-ME cells alone did not cause lymphoma development in pre-irradiated NFS/N mice, while transplantation of both MCF- and ecotropic virus-infected NFS-ME cells resulted in the development of lymphomas of host origin. These results show that lymphomagenic MCF virus was generated through the recombination of E4 viral genome and a modified proviral DNA of endogenous viruses present in radiation- or chemically-induced lymphomas, and that an interaction or synergism of MCF and ecotropic viruses is required for MCF virus to exert lymphomagenic activity. (author)

  10. Hepatorenal protective effect of Antistax ® against chemically-induced toxicity

    Directory of Open Access Journals (Sweden)

    Atallah F Ahmed

    2015-01-01

    Full Text Available Background: Antioxidant natural products and chemoprevention are considered nowadays as an effective approach against health various disorders and diseases induced by oxidative stress or free radicals. Objective: The aim of this study was to assess the hepato- and nephroprotective activity of a standardized red vine leaf aqueous extract AS195 (Antistax ® . Methods: The protective activity of AS195 (100 mg/kg was investigated on carbon tetrachloride (CCl 4 -intoxicated rats in comparison with silymarin. The flavonoid/proanthocyanidin nature of AS195 was identified by phytochemical and nuclear magnetic resonance (NMR analyses, while its total phenol/proanthocyanidin/flavonoid content and antioxidant activity were determined by Folin-Ciocalteau, vanillin-sulfuric acid, AlCl 3 , and 2,2-diphenyl-2-picrylhydrazyl radical scavenging assays, respectively. Results: Relative to the control CCl 4 -intoxicated group, pretreatment with AS195 could significantly suppressed the elevated serum levels of alanine aminotransferase, alkaline phosphatase, γ-glutamyl transferase, total cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, triglycerides, bilirubin, creatinine, uric acid, and calcium, whereas it significantly increased the diminished serum levels of high-density lipoprotein cholesterol, albumin and total protein. Moreover, AS195 significantly decreased malondialdehyde formation in the tissues of liver and kidney, whereas it significantly elevated and nonprotein sulfhydryl groups, compared with the intoxicated control. The improvement in biochemical parameters by AS195 was obviously observed and further confirmed by restoration of normal histological features in the two organs. Conclusions: The results of the present study revealed the capacity of AS195 to enhance the recovery from xenobiotic-induced hepatorenal toxicity initiated by free radicals.

  11. Mechanisms of the hepatoprotective effects of tamoxifen against drug-induced and chemical-induced acute liver injuries

    International Nuclear Information System (INIS)

    Although estrogen receptor (ER)α agonists, such as estradiol and ethinylestradiol (EE2), cause cholestasis in mice, they also reduce the degree of liver injury caused by hepatotoxicants as well as ischemia–reperfusion. The functional mechanisms of ERα have yet to be elucidated in drug-induced or chemical-induced liver injury. The present study investigated the effects of an ERα agonist, selective ER modulators (SERMs) and an ER antagonist on drug-induced and chemical-induced liver injuries caused by acetaminophen, bromobenzene, diclofenac, and thioacetamide (TA). We observed hepatoprotective effects of EE2, tamoxifen (TAM) and raloxifene pretreatment in female mice that were exposed to a variety of hepatotoxic compounds. In contrast, the ER antagonist did not show any hepatoprotective effects. DNA microarray analyses suggested that monocyte to macrophage differentiation-associated 2 (Mmd2) protein, which has an unknown function, is commonly increased by TAM and RAL pretreatment, but not by pretreatment with the ER antagonist. In ERα-knockout mice, the hepatoprotective effects of TAM and the increased expression of Mmd2 mRNA were not observed in TA-induced liver injury. To investigate the function of Mmd2, the expression level of Mmd2 mRNA was significantly knocked down to approximately 30% in mice by injection of siRNA for Mmd2 (siMmd2). Mmd2 knockdown resulted in a reduction of the protective effects of TAM on TA-induced liver injury in mice. This is the first report of the involvement of ERα in drug-induced or chemical-induced liver injury. Upregulation of Mmd2 protein in the liver was suggested as the mechanism of the hepatoprotective effects of EE2 and SERMs. -- Highlights: ► Liver injury induced by drugs or chemicals was investigated in mice. ► Liver injury was suppressed by pretreatment with tamoxifen in female mice. ► Mmd2, whose function was unknown, could be a candidate gene for liver protection. ► Tamoxifen up-regulated Mmd2 mRNA expression

  12. Mechanisms of the hepatoprotective effects of tamoxifen against drug-induced and chemical-induced acute liver injuries

    Energy Technology Data Exchange (ETDEWEB)

    Yoshikawa, Yukitaka; Miyashita, Taishi; Higuchi, Satonori [Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa 920‐1192 (Japan); Tsuneyama, Koichi [Department of Diagnostic Pathology, Graduate School of Medicine and Pharmaceutical Science for Research, University of Toyama, Sugitani, Toyama 930‐0194 (Japan); Endo, Shinya [Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa 920‐1192 (Japan); Tsukui, Tohru [Research Center for Genomic Medicine, Saitama Medical University, Yamane, Hidaka 350‐1241 (Japan); Toyoda, Yasuyuki; Fukami, Tatsuki; Nakajima, Miki [Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa 920‐1192 (Japan); Yokoi, Tsuyoshi, E-mail: tyokoi@p.kanazawa-u.ac.jp [Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa 920‐1192 (Japan)

    2012-10-01

    Although estrogen receptor (ER)α agonists, such as estradiol and ethinylestradiol (EE2), cause cholestasis in mice, they also reduce the degree of liver injury caused by hepatotoxicants as well as ischemia–reperfusion. The functional mechanisms of ERα have yet to be elucidated in drug-induced or chemical-induced liver injury. The present study investigated the effects of an ERα agonist, selective ER modulators (SERMs) and an ER antagonist on drug-induced and chemical-induced liver injuries caused by acetaminophen, bromobenzene, diclofenac, and thioacetamide (TA). We observed hepatoprotective effects of EE2, tamoxifen (TAM) and raloxifene pretreatment in female mice that were exposed to a variety of hepatotoxic compounds. In contrast, the ER antagonist did not show any hepatoprotective effects. DNA microarray analyses suggested that monocyte to macrophage differentiation-associated 2 (Mmd2) protein, which has an unknown function, is commonly increased by TAM and RAL pretreatment, but not by pretreatment with the ER antagonist. In ERα-knockout mice, the hepatoprotective effects of TAM and the increased expression of Mmd2 mRNA were not observed in TA-induced liver injury. To investigate the function of Mmd2, the expression level of Mmd2 mRNA was significantly knocked down to approximately 30% in mice by injection of siRNA for Mmd2 (siMmd2). Mmd2 knockdown resulted in a reduction of the protective effects of TAM on TA-induced liver injury in mice. This is the first report of the involvement of ERα in drug-induced or chemical-induced liver injury. Upregulation of Mmd2 protein in the liver was suggested as the mechanism of the hepatoprotective effects of EE2 and SERMs. -- Highlights: ► Liver injury induced by drugs or chemicals was investigated in mice. ► Liver injury was suppressed by pretreatment with tamoxifen in female mice. ► Mmd2, whose function was unknown, could be a candidate gene for liver protection. ► Tamoxifen up-regulated Mmd2 mRNA expression

  13. Chondroitinase ABC plus bone marrow mesenchymal stem cells for repair of spinal cord injury

    Institute of Scientific and Technical Information of China (English)

    Chun Zhang; Xijing He; Haopeng Li; Guoyu Wang

    2013-01-01

    As chondroitinase ABC can improve the hostile microenvironment and cell transplantation is proven to be effective after spinal cord injury, we hypothesized that their combination would be a more effective treatment option. At 5 days after T8 spinal cord crush injury, rats were injected with bone marrow mesenchymal stem cell suspension or chondroitinase ABC 1 mm from the edge of spinal cord damage zone. Chondroitinase ABC was first injected, and bone marrow mesenchymal stem cell suspension was injected on the next day in the combination group. At 14 days, the mean Basso, Beattie and Bresnahan score of the rats in the combination group was higher than other groups. Hematoxylin-eosin staining showed that the necrotic area was significantly reduced in the combination group compared with other groups. Glial fibrillary acidic protein-chondroitin sulfate proteoglycan double staining showed that the damage zone of astrocytic scars was significantly reduced without the cavity in the combination group. Glial fibrillary acidic protein/growth associated protein-43 double immunostaining revealed that positive fibers traversed the damage zone in the combination group. These results suggest that the combination of chondroitinase ABC and bone marrow mesenchymal stem cell transplantation contributes to the repair of spinal cord injury.

  14. Neuroprotection and immunomodulation by xenografted human mesenchymal stem cells following spinal cord ventral root avulsion.

    Science.gov (United States)

    Ribeiro, Thiago B; Duarte, Adriana S S; Longhini, Ana Leda F; Pradella, Fernando; Farias, Alessandro S; Luzo, Angela C M; Oliveira, Alexandre L R; Olalla Saad, Sara Teresinha

    2015-01-01

    The present study investigates the effects of xenotransplantation of Adipose Tissue Mesenchymal Stem Cells (AT-MSCs) in animals after ventral root avulsion. AT-MSC has similar characteristics to bone marrow mesenchymal stem cells (BM-MSCs), such as immunomodulatory properties and expression of neurotrophic factors. In this study, Lewis rats were submitted to surgery for unilateral avulsion of the lumbar ventral roots and received 5 × 10(5) AT-MSCs via the lateral funiculus. Two weeks after cell administration, the animals were sacrificed and the moto neurons, T lymphocytes and cell defense nervous system were analyzed. An increased neuronal survival and partial preservation of synaptophysin-positive nerve terminals, related to GDNF and BDNF expression of AT-MSCs, and reduction of pro-inflammatory reaction were observed. In conclusion, AT-MSCs prevent second phase neuronal injury, since they suppressed lymphocyte, astroglia and microglia effects, which finally contributed to rat motor-neuron survival and synaptic stability of the lesioned motor-neuron. Moreover, the survival of the injected AT- MSCs lasted for at least 14 days. These results indicate that neuronal survival after lesion, followed by mesenchymal stem cell (MSC) administration, might occur through cytokine release and immunomodulation, thus suggesting that AT-MSCs are promising cells for the therapy of neuronal lesions. PMID:26548646

  15. Inducing human umbilical cord blood mesenchymal stem cells into insulin secreting cells in vitro culture and its effects on diabetes rats%体外培养人脐带血间充质干细胞向胰岛细胞分化及对糖尿病治疗的实验研究

    Institute of Scientific and Technical Information of China (English)

    王英; 罗浩虹; 高洪泉; 缪智辉; 赵亚清

    2014-01-01

    目的:研究人脐带血间充质干细胞向胰岛素分泌细胞分化的潜能及其移植后对糖尿病大鼠的治疗效果。方法体外分离培养HUCB-MSCs,在胰岛细胞培养条件下经药物定向诱导其分化;免疫组化对诱导细胞进行胰岛β细胞标记鉴定;双硫腙染色鉴定锌离子表达及检测胰岛样细胞的移植效果。结果 HUCB-MSCs经诱导后,免疫细胞化学染色显示表达人胰岛素;双硫腙染色呈棕红色;移植后2周,胰岛样细胞组血糖浓度明显降低。结论 HUCB-MSCs在体外诱导培养条件下,具有向胰岛素分泌细胞分化的潜能,这种细胞可能为Ⅰ型糖尿病提供一条新的治疗途径。%Objective To explore the possibility of inducing the differentiation of human umbilical cord blood mesenchymal stem cells(HUCB-MSCs) into insulin secreting cells in vitro and the therapeutic efficacy on diabetic rats following transplantation.Methods We isolated and cultured HUCB-MSCs, and induced HUCB-MSCs to differentiate into insulin secreting cells in the islet cell culture condition. HUCB-MSCs of induction were detected by immunocytochemical Methods. Zinchydronium was detected by Dithizon staining. Transplantation efficiency of islet-like cells was detected.Results Under induction, immunocytochemical examination showed that the expression of human insulin was positive. Dithizon stain showed that the cytoplasm of HUCB-MSCs was stained in brownish red color after induction. At 2 weeks following transplantation, blood glucose concentration in the islet-like cell group significantly reduced.Conclusion HUCB-MSCs can be differentiated into insulin secreting cells in vitro. HUCB-MSCs have the potential to become an excellent candidate in β cell replacement therapy of typeⅠdiabetes.

  16. Human umbilical cord matrix-derived stem cells exert trophic effects on β-cell survival in diabetic rats and isolated islets

    Directory of Open Access Journals (Sweden)

    Yunting Zhou

    2015-12-01

    Full Text Available Human umbilical cord matrix-derived stem cells (uMSCs, owing to their cellular and procurement advantages compared with mesenchymal stem cells derived from other tissue sources, are in clinical trials to treat type 1 (T1D and type 2 diabetes (T2D. However, the therapeutic basis remains to be fully understood. The immunomodulatory property of uMSCs could explain the use in treating T1D; however, the mere immune modulation might not be sufficient to support the use in T2D. We thus tested whether uMSCs could exert direct trophic effects on β-cells. Infusion of uMSCs into chemically induced diabetic rats prevented hyperglycemic progression with a parallel preservation of islet size and cellularity, demonstrating the protective effect of uMSCs on β-cells. Mechanistic analyses revealed that uMSCs engrafted long-term in the injured pancreas and the engraftment markedly activated the pancreatic PI3K pathway and its downstream anti-apoptotic machinery. The pro-survival pathway activation was associated with the expression and secretion of β-cell growth factors by uMSCs, among which insulin-like growth factor 1 (IGF1 was highly abundant. To establish the causal relationship between the uMSC-secreted factors and β-cell survival, isolated rat islets were co-cultured with uMSCs in the transwell system. Co-culturing improved the islet viability and insulin secretion. Furthermore, reduction of uMSC-secreted IGF1 via siRNA knockdown diminished the protective effects on islets in the co-culture. Thus, our data support a model whereby uMSCs exert trophic effects on islets by secreting β-cell growth factors such as IGF1. The study reveals a novel therapeutic role of uMSCs and suggests that multiple mechanisms are employed by uMSCs to treat diabetes.

  17. Mesenchymal Stem Cell-Based Therapy

    OpenAIRE

    Mundra, Vaibhav; Gerling, Ivan C.; Mahato, Ram I.

    2012-01-01

    Mesenchymal stem cells (MSCs) are multipotent adult stem cells which have self-renewal capacity and differentiation potential into several mesenchymal lineages including bones, cartilages, adipose tissues and tendons. MSCs may repair tissue injuries and prevent immune cell activation and proliferation. Immunomodulation and secretion of growth factors by MSCs have led to realizing the true potential of MSC-based cell therapy. The use of MSCs as immunomdulators has been explored in cell/organ t...

  18. Mesenchymal Cystic Hamartoma Presenting with Spontaneous Pneumothorax

    OpenAIRE

    J Glezos; Toppin, D; Cooney, T

    2003-01-01

    A 53-year-old woman presented with a spontaneous right-sided pneumothorax. Computed tomography chest scan demonstrated a large bulla involving the right middle lobe. Recurrence of the pneumothorax after initial closed chest tube drainage necessitated thoracotomy and wedge resection. Histopathological examination revealed features consistent with a benign mesenchymal cystic hamartoma. Pneumothorax is a recognized complication of mesenchymal cystic hamartoma, a lesion that also has malignant po...

  19. Pancreatic Mesenchyme Regulates Epithelial Organogenesis throughout Development

    OpenAIRE

    Landsman, Limor; Nijagal, Amar; Whitchurch, Theresa J.; VanderLaan, Renee L.; Zimmer, Warren E.; MacKenzie, Tippi C.; Hebrok, Matthias

    2011-01-01

    The developing pancreatic epithelium gives rise to all endocrine and exocrine cells of the mature organ. During organogenesis, the epithelial cells receive essential signals from the overlying mesenchyme. Previous studies, focusing on ex vivo tissue explants or complete knockout mice, have identified an important role for the mesenchyme in regulating the expansion of progenitor cells in the early pancreas epithelium. However, due to the lack of genetic tools directing expression specifically ...

  20. Mesenchymal stem cells in regenerative rehabilitation

    OpenAIRE

    Nurkovic, Jasmin; Dolicanin, Zana; Mustafic, Fahrudin; Mujanovic, Rifat; Memic, Mensur; Grbovic, Vesna; Skevin, Aleksandra Jurisic; Nurkovic, Selmina

    2016-01-01

    [Purpose] Regenerative medicine and rehabilitation contribute in many ways to a specific plan of care based on a patient’s medical status. The intrinsic self-renewing, multipotent, regenerative, and immunosuppressive properties of mesenchymal stem cells offer great promise in the treatment of numerous autoimmune, degenerative, and graft-versus-host diseases, as well as tissue injuries. As such, mesenchymal stem cells represent a therapeutic fortune in regenerative medicine. The aim of this re...

  1. Mesenchymal stem cells in regenerative rehabilitation

    Science.gov (United States)

    Nurkovic, Jasmin; Dolicanin, Zana; Mustafic, Fahrudin; Mujanovic, Rifat; Memic, Mensur; Grbovic, Vesna; Skevin, Aleksandra Jurisic; Nurkovic, Selmina

    2016-01-01

    [Purpose] Regenerative medicine and rehabilitation contribute in many ways to a specific plan of care based on a patient’s medical status. The intrinsic self-renewing, multipotent, regenerative, and immunosuppressive properties of mesenchymal stem cells offer great promise in the treatment of numerous autoimmune, degenerative, and graft-versus-host diseases, as well as tissue injuries. As such, mesenchymal stem cells represent a therapeutic fortune in regenerative medicine. The aim of this review is to discuss possibilities, limitations, and future clinical applications of mesenchymal stem cells. [Subjects and Methods] The authors have identified and discussed clinically and scientifically relevant articles from PubMed that have met the inclusion criteria. [Results] Direct treatment of muscle injuries, stroke, damaged peripheral nerves, and cartilage with mesenchymal stem cells has been demonstrated to be effective, with synergies seen between cellular and physical therapies. Over the past few years, several researchers, including us, have shown that there are certain limitations in the use of mesenchymal stem cells. Aging and spontaneous malignant transformation of mesenchymal stem cells significantly affect the functionality of these cells. [Conclusion] Definitive conclusions cannot be made by these studies because limited numbers of patients were included. Studies clarifying these results are expected in the near future. PMID:27390452

  2. Cobalt surface modification during γ-Fe{sub 2}O{sub 3} nanoparticle synthesis by chemical-induced transition

    Energy Technology Data Exchange (ETDEWEB)

    Li, Junming [School of Physical Science and Technology, Southwest University, Chongqing 400715 (China); Li, Jian, E-mail: aizhong@swu.edu.cn [School of Physical Science and Technology, Southwest University, Chongqing 400715 (China); Chen, Longlong; Lin, Yueqiang; Liu, Xiaodong; Gong, Xiaomin [School of Physical Science and Technology, Southwest University, Chongqing 400715 (China); Li, Decai [School of Mechanical and Control Engineering, Beijing Jiaotong University, Beijing 100044 (China)

    2015-02-01

    In the chemical-induced transition of FeCl{sub 2} solution, the FeOOH/Mg(OH){sub 2} precursor was transformed into spinel structured γ-Fe{sub 2}O{sub 3} crystallites, coated with a FeCl{sub 3}·6H{sub 2}O layer. CoCl{sub 2} surface modified γ-Fe{sub 2}O{sub 3} nanoparticles were prepared by adding Co(NO{sub 3}){sub 2} during the synthesis. CoFe{sub 2}O{sub 4} modified γ-Fe{sub 2}O{sub 3} nanoparticles were prepared by adding NaOH during the surface modification with Co(NO{sub 3}){sub 2}. The CoFe{sub 2}O{sub 4} layer grew epitaxially on the γ-Fe{sub 2}O{sub 3} crystallite to form a composite crystallite, which was coated by CoCl{sub 2}·6H{sub 2}O. The composite could not be distinguished using X-ray diffraction or transmission electron microscopy, since CoFe{sub 2}O{sub 4} and γ-Fe{sub 2}O{sub 3} possess similar spinel structures and lattice constants. X-ray photoelectron spectroscopy was used to distinguish them. The saturation magnetization and coercivity of the spinel structured γ-Fe{sub 2}O{sub 3}-based nanoparticles were related to the grain size. - Highlights: • γ-Fe{sub 2}O{sub 3} nanoparticles were synthesized by chemical induced transition. • CoCl{sub 2} modified nanoparticles were prepared by additional Co(NO{sub 3}){sub 2} during synthesization. • CoFe{sub 2}O{sub 4} modified nanoparticles were prepared by additional Co(NO{sub 3}){sub 2} and NaOH. • The magnetism of the nanoparticles is related to the grain size.

  3. Sensitivity of neuroprogenitor cells to chemical-induced apoptosis using a multiplexed assay suitable for high-throughput screening

    International Nuclear Information System (INIS)

    High-throughput methods are useful for rapidly screening large numbers of chemicals for biological activity, including the perturbation of pathways that may lead to adverse cellular effects. In vitro assays for the key events of neurodevelopment, including apoptosis, may be used in a battery of tests for detecting chemicals that could result in developmental neurotoxicity. Apoptosis contributes to nervous system development by regulating the size of the neuroprogenitor cell pool, and the balance between cellular proliferation and apoptosis during neuroprogenitor cell proliferation helps to determine the size and shape of the nervous system. Therefore, chemicals that affect apoptosis during neuronal development can have deleterious effects on the developing brain. The present study examined the utility of a high-throughput assay to detect chemical-induced apoptosis in mouse or human neuroprogenitor cells, as well as differentiated human neurons derived from induced pluripotent stem cells. Apoptosis was assessed using an assay that measures enzymatic activity of caspase-3/7 in a rapid and cost efficient manner. The results show that all three commercially available models generated a robust source of proliferating neuroprogenitor cells, and that the assay was sensitive and reproducible when used in a multi-well plate format. There were differences in the response of rodent and human neuroprogenitor cells to a set of chemicals previously shown to induce apoptosis in vitro. Neuroprogenitor cells were more sensitive to chemical-induced apoptosis than differentiated neurons, suggesting that neuroprogenitor cells are one of the cell models that should be considered for use in a developmental neurotoxicity screening battery

  4. Lipopolysaccharide-activated microglial-induced neuroglial cell differentiation in bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    Xiaoguang Luo; Chunlin Ge; Yan Ren; Hongmei Yu; Zhe Wu; Qiushuang Wang; Chaodong Zhang

    2008-01-01

    BACKGROUND: Microglia are very sensitive to environmental changes, often becoming activated by pathological conditions. Activated microglia can exert a dual role in injury and repair in various diseases of the central nervous system, including cerebral ischemia, Parkinson's disease, and Alzheimer's disease. OBJECTIVE: An immortal microglial cell line, BV2, was treated with varying concentrations of lipopolysaccharide (LPS) to induce a pathological situation. Supernatant was harvested and incubated with bone marrow mesenchymal stem cells and, concomitantly, bone marrow mesenchymal stem cell differentiation was observed. DESIGN: A controlled observation, in vitro experiment. SETTING: Department of Neurology, First Affiliated Hospital of China Medical University. MATERIALS: Five male 2-3-week-old Sprague Dawley rats were purchased from Animal Laboratory Center of China Medical University and included in this study. The protocol was performed in accordance with ethical guidelines for the use and care of animals. The microglial cell line BV2 was produced by Cell Research Institute of Chinese Academy of Sciences. LPS was produced by Sigma Company, USA. METHODS: This study was performed in the Central Laboratory of China Medical University from September 2006 to March 2007. Rat femoral and tibial bone marrow was collected for separation and primary culture of bone marrow mesenchymal stem cells. Bone marrow mesenchymal stem cell cultures were divided into 5 groups: control group, non-activated group, as well as low-, medium-, and high-dose LPS groups. In the control group, bone marrow mesenchymal stem cells were cultured with Dulbecco's modified Eagle's medium (DMEM) supplemented with fetal bovine serum (volume fraction 0.1). In the non-activated group, bone marrow mesenchymal stem cells were incubated with non-activated BV2 supernatant. In the low-, medium-, and high-dose LPS groups, bone marrow mesenchymal stem cells were incubated with LPS (0.01, 0.1 and 1

  5. Light-induced retinal injury enhanced neurotrophins secretion and neurotrophic effect of mesenchymal stem cells in vitro

    Directory of Open Access Journals (Sweden)

    Wei Xu

    2013-04-01

    Full Text Available PURPOSE: To investigate neurotrophins expression and neurotrophic effect change in mesenchymal stem cells (MSCs under different types of stimulation. METHODS: Rats were exposed in 10,000 lux white light to develop light-induced retinal injury. Supernatants of homogenized retina (SHR, either from normal or light-injured retina, were used to stimulate MSCs. Quantitative real time for polymerase chain reaction (RT-PCR and enzyme-linked immunosorbent assay (ELISA were conducted for analysis the expression change in basic fibroblast growth factor (bFGF, brain-derived neurotrophic factor (BDNF and ciliary neurotrophic factor (CNTF in MSCs after stimulation. Conditioned medium from SHR-stimulated MSCs and control MSCs were collected for evaluation their effect on retinal explants. RESULTS: Supernatants of homogenized retina from light-injured rats significantly promoted neurotrophins secretion from MSCs (p<0.01. Conditioned medium from mesenchymal stem cells stimulated by light-injured SHR significantly reduced DNA fragmentation (p<0.01, up-regulated bcl-2 (p<0.01 and down-regulated bax (p<0.01 in retinal explants, displaying enhanced protective effect. CONCLUSIONS: Light-induced retinal injury is able to enhance neurotrophins secretion from mesenchymal stem cells and promote the neurotrophic effect of mesenchymal stem cells.

  6. Cadmium affects viability of bone marrow mesenchymal stem cells through membrane impairment, intracellular calcium elevation and DNA breakage

    Directory of Open Access Journals (Sweden)

    Abnosi Mohammad Hussein

    2010-01-01

    Full Text Available Background: Cadmium is an important heavy metal with occupational and environmental hazard. Cadmium toxicity results mainly in bone-related complication such as itai-itai disease. Mesenchymal stem cells of the bone marrow have the ability to differentiate to osteoblasts which ensure the well-being of the bone tissue. Thus the aim was to investigate the effect of cadmium on viability of rat bone marrow mesenchymal stem cells. Materials and Methods: The rat bone marrow mesenchymal stem cells were grown to confluency in DMEM medium supplemented with 15% fetal bovine serum and penicillin-streptomycin up to third passage. Then the cells were treated with 0, 5, 15, 25, 35, and 45 of CdCl 2 at 12, 24, 36, and 48 h, and their viability was investigated using trypan blue staining. In addition, after treatment with selected dose (15 and 45 μM and time (24 and 48 h the cell morphology, DNA damage and calcium content of the cells were evaluated. Data was analyzed using one and two-way ANOVA (Tukey test and the P2+ was observed. Conclusion: Cadmium chloride is a toxic compound which might affect the well-being of bone tissue through affecting the mesenchymal stem cells.

  7. Study on therapeutical effect of tracheal injection mesenchymal stem cells on pulmonary arterial hypertension in rats%经气管注射骨髓间充质干细胞治疗早期血管炎性作用所致肺动脉高压的实验研究

    Institute of Scientific and Technical Information of China (English)

    曲素萍; 牛丽丽

    2012-01-01

    Objective To discuss the effect and therapeutic mechanism of bone marrow mesenchymal stem cells ( MSCs ) transpl antation on the animal model of pulmonary arterial hypertension ( PAH ) and observe the changes of vessels which transfordmed with MSCs on the animal model. Methods The rats were randomly divided into four groups: the blank control group with normal saline ( Con group ), the non-treatment group ( PAH group ), the vein-al treatment group ( VMSCs group ) and tracheal treatment group ( TMSCs group ). The PAH model was induced by subcutaneous injection of MCT in VMSCs, TMSCs and PAH groups, the rats of the control group were injected with normal saline. One week after MCT administration, MSCs were injected through the ranine vein or tracheal into rats in the VMSCs and TMSCs groups. At the same time, rats in PAH and Con groups were injected with the same a-mount of low-glucose DMEM ( L-DMEM ). Three weeks after the injection of MCT, the pulmonary artery pressure and the expression of Smad2 and its active form were tested. The thickness of vascular wall was observed. The fluorescence's distribution in the lung tissue in TMSCs group and VMSCs group in 10 days and 21 days after the injection of MCT were observed. Results When the SD rats were injected with the MCT for 21 days, the SPAP, MPAP in VMSCs and TMSCs groups declined obviously compared with PAH group ( P < 0. 05 ), but higher than the Con group ( P < 0. 05 ). VMSCs and TMSCs groups vessel wall thickness was thinner than PAH group ( P < 0. 05 ). The level of Smad2 in PAH group, TMSCs group, VMSCs group was with no significant differences, but slightly higher than Con group; the level of Smad2 phosphorylation in PAH group was higher than VMSCs group, TMSCs group, Con group. SPAP, MPAP, vessel wall thickness, Smad2 and Smad2 phosphorylation in VMSCs and TMSCs groups showed no sigrificant different. Conclusion MSCs treatment reduce the pressure of the vessels and the function of reconstruction, which may be

  8. 多种诱导剂联合对大鼠骨髓间充质干细胞体外向神经元样细胞分化的影响%Influence of combined various inducers on rat mesenchymal stem cells differentiate into neuronal-like phenotype

    Institute of Scientific and Technical Information of China (English)

    陈翔; 樊兴娟; 毛伟峰

    2012-01-01

    Objective To explore the influence of combined various inducers on rat mesenchymal stem cells (rMSCs) differentiate into neuronal-like phenotype. Methods The rMSCs were separated from Wistar rat's bone marrow and cultured in vitro. After appraisal by immunofluorescence staining and flow cytometry instrument line CD44, CD45 and CD90, the third generation of rMSCs were pr-induced by DMEM, 0. 01 mmol/L BME and 2% DMSO for 5 h; then they were combine induced by 10 μg/L basic fibroblast growth factor (bFGF), 10 μg/L epidermal growth factor (EGF) , and 10 μ/L creatine for 7 d. Cells morphology was observed by fluorescent phase contrast microscope, and neuronal markers were detected using immumohistochemistry and flow cytometry. Results About 80% rMSCs were exhibited a neuron model change after induced. In those cells the positive rate of neuronal markers in Nestin was (42. 14 ±2. 85)% , nerve filament protein was (17. 13 ± 1. 03)% , β-tubulin M was (20.42 ± 1. 98) % , gum fibre acid protein was (39. 97 ± 2. 34) % , microtubule associated protein-2 was (39. 30 ± 1.72)% , choline acetyl transferase was (21. 30 ± 1. 83)% respectively. Conclusions Combined with various inducers (BME, DMSO, bFGF, EGF and creatine) , about 80% rMSCs can be induced to neuronal-like cells.%目的 探讨多种诱导剂联合对大鼠骨髓间充质干细胞(rMSCs)体外向神经元样细胞分化的影响.方法 从Wistar大鼠骨髓中分离rMSCs并进行体外培养,经免疫荧光染色及流式细胞仪行CD44、CD45和CD90鉴定后,将传第3代的rMSCs用DMEM、0.1 mmol/L β-巯基乙醇(BME)和2%二甲基亚砜(DMSO)预诱导5h后,给予10μg/L碱性成纤维细胞生长因子(bFGF)、10 μg/L表皮生长因子(EGF)、10 μg/L肌酸联合诱导7d.应用荧光相差显微镜观察细胞形态,免疫荧光染色和流式细胞仪对神经元标志物进行鉴定.结果 诱导后约80% rMSCs呈神经元样改变,神经元标志物阳性率分别是巢蛋白为(42.14±2.85)

  9. Distribution of Mesenchymal Stem Cells and Effects on Neuronal Survival and Axon Regeneration after Optic Nerve Crush and Cell Therapy

    OpenAIRE

    Mesentier-Louro, Louise Alessandra; Zaverucha-do-Valle, Camila; da Silva-Junior, Almir Jordão; Nascimento-dos-Santos, Gabriel; Gubert, Fernanda; de Figueirêdo, Ana Beatriz Padilha; Torres, Ana Luiza; Paredes, Bruno D; Teixeira, Camila; Tovar-Moll, Fernanda; Mendez-Otero, Rosalia; MARCELO F. SANTIAGO

    2014-01-01

    Bone marrow-derived cells have been used in different animal models of neurological diseases. We investigated the therapeutic potential of mesenchymal stem cells (MSC) injected into the vitreous body in a model of optic nerve injury. Adult (3–5 months old) Lister Hooded rats underwent unilateral optic nerve crush followed by injection of MSC or the vehicle into the vitreous body. Before they were injected, MSC were labeled with a fluorescent dye or with superparamagnetic iron oxide nanopartic...

  10. Hypoxia-induced alveolar epithelial-mesenchymal transition requires mitochondrial ROS and hypoxia-inducible factor 1

    OpenAIRE

    Zhou, Guofei; Dada, Laura A.; Wu, Minghua; Kelly, Aileen; Trejo, Humberto; Zhou, Qiyuan; Varga, John; Sznajder, Jacob I.

    2009-01-01

    Patients with acute lung injury develop hypoxia, which may lead to lung dysfunction and aberrant tissue repair. Recent studies have suggested that epithelial-mesenchymal transition (EMT) contributes to pulmonary fibrosis. We sought to determine whether hypoxia induces EMT in alveolar epithelial cells (AEC). We found that hypoxia induced the expression of α-smooth muscle actin (α-SMA) and vimentin and decreased the expression of E-cadherin in transformed and primary human, rat, and mouse AEC, ...

  11. Effects of Negative Pressure Wound Therapy on Mesenchymal Stem Cells Proliferation and Osteogenic Differentiation in a Fibrin Matrix

    OpenAIRE

    Jin Zhu; Aixi Yu; Baiwen Qi; Zonghuan Li; Xiang Hu

    2014-01-01

    Vacuum-assisted closure (VAC) negative pressure wound therapy (NPWT) has been proven to be an effective therapeutic method for the treatment of recalcitrant wounds. However, its role in bone healing remains to be unclear. Here, we investigated the effects of NPWT on rat periosteum-derived mesenchymal stem cells (P-MSCs) proliferation and osteoblastic differentiation in a 3D fibrin matrix. P-MSCs underwent primary culture for three passages before being used to construct cell clots. The fibrin...

  12. Osteogenic differentiation of mesenchymal stem cells on demineralized and devitalized biodegradable polymer and extracellular matrix hybrid constructs

    OpenAIRE

    Thibault, Richard A.; Mikos, Antonios G.; Kasper, F. Kurtis

    2013-01-01

    Devitalization and demineralization processing of biodegradable polymer and extracellular matrix (ECM) hybrid constructs was explored for the effect on the retention of ECM components and construct osteogenicity. Hybrid constructs were generated by seeding osteogenically pre-differentiated rat mesenchymal stem cells (MSCs) onto electrospun poly(ε-caprolactone) fiber meshes and culturing in osteogenic medium for 12 or 16 days within a flow perfusion bioreactor to create an ECM coating. The res...

  13. Concentrated Hypoxia-Preconditioned Adipose Mesenchymal Stem Cell-Conditioned Medium Improves Wounds Healing in Full-Thickness Skin Defect Model.

    Science.gov (United States)

    Sun, Biao; Guo, Shilei; Xu, Fei; Wang, Bin; Liu, Xiujuan; Zhang, Yuanyuan; Xu, Yan

    2014-01-01

    In recent years, the bioactive factors were utilized in exercise and athletic skin injuries. In this research, the concentrated conditioned medium of hypoxia-preconditioned adipose mesenchymal stem cells, which is rich in bioactive factor, is applied in full-thickness skin defect model to evaluate the therapeutic efficacy. Adipose mesenchymal stem cells were harvested from the abdominal subcutaneous adipose tissues. The surface markers and the potential of differentiation were analyzed. The conditioned medium of hypoxia-preconditioned stem cells was collected and freeze-dried and then applied on the rat full-thickness skin defect model, and the healing time of each group was recorded. Haematoxylin and eosin staining of skin was assessed by microscope. The characteristics of adipose mesenchymal stem cells were similar to those of other mesenchymal stem cells. The concentration of protein in freeze-dried conditioned medium in 1 mL water was about 15 times higher than in the normal condition medium. In vivo, the concentrated hypoxia-preconditioned conditioned medium can reduce the wound size and accelerate the skin wound healing. The concentrated hypoxia-preconditioned adipose mesenchymal stem cell-conditioned medium has great effect on rat model of wound healing, and it would be an ideal agent for wound care in clinical application. PMID:27433483

  14. 骨髓间充质干细胞、肌样细胞藻酸钙复合凝胶在压力性尿失禁大鼠尿道周围的成肌效应研究%The effects of the gel compound from bone marrow mesenchymal stem cells and muscle-like cells/calcium alginate on myoblast formation around urethra in rats of stress urinary incontinence

    Institute of Scientific and Technical Information of China (English)

    杜小文; 吴慧玲; 朱永锋; 胡俊彪; 金范; 吕蕊萍; 徐剑炜; 孙思; 王皓羽

    2012-01-01

    目的 探索骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)藻酸钙复合凝胶与经5-氮杂胞苷诱导的肌样细胞藻酸钙复合凝胶在压力性尿失禁(stress urinary incontinence,SUI)大鼠尿道周围的成肌效应.方法 SD大鼠BMSC体外分离、培养、鉴定;用5-氮杂胞苷诱导生成肌样细胞;2%藻酸钠与1%氯化钙溶液以5∶1体积比配制藻酸钙凝胶,分别与BMSC和肌样细胞复合用于微量注射.72只6周龄雌性SD大鼠建立SUI模型后分为BMSC凝胶组、肌样细胞凝胶组、单纯凝胶组和空白对照4组,每组再分为3小组,于膀胱颈尿道移行部黏膜下肌层注射相应的复合凝胶.4周、8周时取各组大鼠尿道横截面进行HE染色、荧光示踪照相以及结蛋白、α-横纹肌动蛋白(α-SMA)染色检查. 结果 获得的BMSC第3代细胞表面细胞因子抗体CD29阳性率为89.4%、CD34为3.3%、CD45为2.5%、CD105为46.0%,荧光示踪照相见第3代培养细胞表达强绿色荧光.BMSC凝胶组和肌样细胞凝胶组4周、8周时,凝胶边缘血管长入并逐渐增多,荧光示踪BMSC聚集于新生血管周围,肌样细胞呈长梭形样生长,结蛋白、d-SMA明显阳性表达. 结论 BMSC、肌样细胞藻酸钙复合凝胶在大鼠SUI实验模型的微环境中具有向肌细胞分化的潜力.BMSC直接体内微环境诱导分化与5-氮杂胞苷体外诱导形成肌样细胞后植入体内微环境继续分化,短期结果无明显差异.%Objective To explore the effects of myoblast formation around the urethra of stress urinary incontinence (SUI) rats after treated with bone marrow mesenchymal stem cells(BMSCs) or musclelike cells/calcium alginate composite gel injection therapy. Methods Isolation,cultivation and identification of Sprague-Dawley rat bone marrow mesenchymal stem cell were performed.5-azacytidine was introduced to induce muscle-like cells.Calcium alginate gel was initially prepared by 2% sodium alginate and 1

  15. Effects of bone marrow mesenchymal stem cells on learning and memory functional recovery in neonatal rats with hypoxic-ischemic brain damage%骨髓间充质干细胞对缺氧缺血性脑损伤新生大鼠学习记忆功能的重建作用

    Institute of Scientific and Technical Information of China (English)

    刘洋; 张萱; 代英; 舒畅; 瞿平; 刘友学; 杨莉; 李廷玉

    2008-01-01

    Objective Neonatal hypoxic-ischemic brain damage(HIBD)causes acute death and chronic nervous system sequelae in newborn infants and children.Whereas there have been no specific treatment towards it up to now.Studies have shown that bone marrow mesenchymal stem cells(MSCS)have the therapeutic potential in many nervous system diseases and the authors previously found that retinoid acid (RA),which plays an important role in brain development,could enhance the neural differentiation of rat MSCs(rMSCs)in vitro.This study aimed to examine effects of rMSCs and RA-preindueed rMSC on learning and memory funotional recovery after HIBD in neonatal rats in order to explore a new treatment strategy for clinical application.and explore the mechanism of action of rMSCs.Methods Rat MSCs were isobted and purified from the whole bone marrow of iuvenile Wistar rats by removing the non-adherent cells in primary and passage cultures.Neonatal hypoxic-ischemic brain damage rat models were built according to the methods described by Rice:the right carotid artery of 7-day-postnatal Wistar rats was ligated under anesthesia,and then the rats were exposed to 8%-9% O2 in a container.At 5 days after hypoxia-ischemia,the HIBD neonatal rats were randomly divided into 3 groups and respectively transplanted with saline.BrdU marked rMSCs (1-2 × 105) or RA-preinduced rMSCs (1-2 × 105) into their lateral cerebral ventricle.Immunohistochemistry for nestin,neuron-specific enolase ( NSE),neurofilament protein-heavy chain ( NF-H) and glial fibrillary acidic protein (GFAP) were used to identify cells derived from rMSCs at 14 days and 42 days after transplantation. Shuttle box test was performed to evaluate the condition of learning and memory functional recovery when animals were 7 weeks old. Neurotrophin and receptors cDNA micrearray were also employed at 14 days after transplantation to investigate the underlying action mechanisms of rMSCs treatment. Real-time PCR was used to confirm some of the

  16. 磁性/荧光量子点双功能纳米粒子双标记对大鼠骨髓间充质干细胞生物学特性的影响%Biological characteristics of rat bone mesenchymal stem cells after magnetic/luminescent quantum dots bifunctional nanoparticles labeling

    Institute of Scientific and Technical Information of China (English)

    颜荣华; 王劲; 单鸿; 聂立波; 张黎明

    2011-01-01

    目的:利用磁性/荧光量子点双功能纳米粒子双标记大鼠骨髓间充质干细胞( BMSOs),探讨其对BMSCs生物学特性的影响.方法:分离、纯化及培养大鼠BMSCs.制备二氧化硅( SiO2)包裹的含四氧化三铁(Fe3O4)和碲化镉(CdTe)荧光量子点的磁性及荧光双功能纳米粒子Fe3O4@ CdTe@ SiO2,利用铁浓度为25 mg/L的Fe3O4@ CdTe@ SiO2双标记BMSCs,未标记的BMSCs作为对照组.采用细胞计数(CCK-8)试剂盒检测BMSCs细胞毒性和增殖能力,台盼蓝拒染测细胞活力,成骨、成脂诱导检测细胞多向分化能力,评价双标记对BMSCs生物学特性的影响.荧光显微镜和普鲁士蓝染色观察诱导分化后Fe3O4@ CdTe@ SiO2双标记情况.结果:荧光显微镜下可见双标记组细胞内Fe3O4@ CdTe@ SiO2纳米粒子显示为红色荧光,荧光和磁性双标记率均达到90%以上.Fe3O4@ CdTe@ SiO2双标记后细胞存活率为(94±5)%.Fe3O4@CdTe@ SiO2对BMSCs无明显细胞毒性,对BMSCs的成脂、成骨分化潜能无明显影响.结论:磁性/荧光量子点双功能纳米粒子(Fe3O4@CdTe@ SiO2纳米粒子)双标记对大鼠BMSCs安全有效,对BMSCs的生物学特性无明显影响,有望为MRI和光学成像活体示踪BMSCs提供技术基础.%Objective; To evaluate the safety of magnetic and luminescent quantum dots bifunctional nanoparti-cles dual-labeling in rat bone mesenchymal stem cells (BMSCs). Methods; Rat BMSCs were isolated, purified, and expanded. Magnetic/luminescent bifunctional nanoparticles (Fe3O4@ CdTe@ SiO2) , were prepared by simultaneous encapsulation of Fe3O4 and CdTe quantum dots with silicon dioxide (SiO2). Subsequently, BMSCs were incubated with the Fe3O4@ CdTe@SiO2 nanoparticles with iron concentration of 25 mg/L The viability, cytotoxici-ty and proliferation activity of dual-labeled BMSCs were evaluated with CCK-8 and trypan blue stain exclusion, respectively. Multilineage differentiation potential of dual-labeled BMSCs was assessed by induced

  17. Immunological characteristics of mesenchymal stem cells

    Directory of Open Access Journals (Sweden)

    Cíntia de Vasconcellos Machado

    2013-01-01

    Full Text Available Although bone marrow is the main source, mesenchymal stem cells have already been isolated from various other tissues, such as the liver, pancreas, adipose tissue, peripheral blood and dental pulp. These plastic adherent cells are morphologically similar to fibroblasts and have a high proliferative potential. This special group of cells possesses two essential characteristics: self-renewal and differentiation, with appropriate stimuli, into various cell types. Mesenchymal stem cells are considered immunologically privileged, since they do not express costimulatory molecules, required for complete T cell activation, on their surface. Several studies have shown that these cells exert an immunosuppressive effect on cells from both innate and acquired immunity systems. Mesenchymal stem cells can regulate the immune response in vitro by inhibiting the maturation of dendritic cells, as well as by suppressing the proliferation and function of T and B lymphocytes and natural killer cells. These special properties of mesenchymal stem cells make them a promising strategy in the treatment of immune mediated disorders, such as graft-versus-host disease and autoimmune diseases, as well as in regenerative medicine. The understanding of immune regulation mechanisms of mesenchymal stem cells, and also those involved in the differentiation of these cells in various lineages is primordial for their successful and safe application in different areas of medicine.

  18. Resident mesenchymal cells and fibrosis✩

    Science.gov (United States)

    Hutchison, Nicol; Fligny, Cécile; Duffield, Jeremy S.

    2013-01-01

    Fibrosis is a major clinical problem associated with as many as 45% of all natural deaths in developed nations. It can affect all organs and accumulating evidence indicates that fibrogenesis is not merely a bystander product of injury, but is a central pathological problem directly contributing to loss of organ function. In the majority of clinical cases, fibrogenesis is strongly associated with the recruitment of leukocytes, even in the absence of infection. Although chronic infections are a significant cause of fibrogenesis, in most cases fibrotic disease occurs in the context of sterile injury, such as microvascular disease, toxic epithelial injury or diabetes mellitus. Fibrogenesis is a direct consequence of the activation of extensive, and previously poorly appreciated, populations of mesenchymal cells in our organs which are either wrapped around capillaries and known as ‘pericytes’, or embedded in interstitial spaces between cell structures and known as resident ‘fibroblasts’. Recent fate-mapping and complementary studies in several organs indicate that these cells are the precursors of the scar-forming myofibroblasts that appear in our organs in response to injury. Here we will review the literature supporting a central role for these cells in fibrogenesis, and highlight some of the critical cell to cell interactions that are necessary for the initiation and continuation of the fibrogenic process. This article is part of a Special Issue entitled: Fibrosis: Translation of basic research to human disease. PMID:23220259

  19. Mesenchymal Stem Cells in Cardiology.

    Science.gov (United States)

    White, Ian A; Sanina, Cristina; Balkan, Wayne; Hare, Joshua M

    2016-01-01

    Cardiovascular disease (CVD) accounts for more deaths globally than any other single disease. There are on average 1.5 million episodes of myocardial infarction (heart attack) each year in the United States alone with roughly one-third resulting in death. There is therefore a major need for developing new and effective strategies to promote cardiac repair. Intramyocardial transplantation of mesenchymal stem cells (MSCs) has emerged as a leading contender in the pursuit of clinical intervention and therapy. MSCs are potent mediators of cardiac repair and are therefore an attractive tool in the development of preclinical and clinical trials. MSCs are capable of secreting a large array of soluble factors, which have had demonstrated effects on pathogenic cardiac remolding, fibrosis, immune activation, and cardiac stem cell proliferation within the damaged heart. MSCs are also capable of differentiation into cardiomyocytes, endothelial cells, and vascular smooth muscle cells, although the relative contribution of trilineage differentiation and paracrine effectors on cardiac repair remains the subject of active investigation. PMID:27236666

  20. Primary malignant liver mesenchymal tumor: A case report

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Primary malignant liver mesenchymal tumor is a rare condition defined as a tumor with vascular, fibrous, adi-pose, and other mesenchymal tissue differentiation. We report a case of primary malignant liver mesenchymal tumor in a 51-year-old male with anemia, weight loss and hepatomegaly. Finally unconventional liver biopsy and histological manifestation led to the definitive diag-nosis.

  1. 电磁场作用下大鼠骨髓间充质干细胞的成骨分化与增殖%Effects of exposure to electromagnetic fields on osteogenic differentiation and proliferation of rats' bone marrow mesenchymal stem cells

    Institute of Scientific and Technical Information of China (English)

    朱波; 吴华; 黄珊珊; 赵东明

    2015-01-01

    Objective To study the effects of electromagnetic field (EMF) on the activation of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) pathways in mesenchymal stem cells (MSCs) and their interaction, and to explore the cellular signal transduction mechanism of the biological effects induced by EMF.Methods The 3rd-passage rat bone marrow MSCs were randomly divided into a control group, an EMF group, an EMF + PD98059 group and an EMF + SB202190 group.Cells in the EMF group were cultured in the electromagnetic field, those in the EMF + PD98059 and EMF + SB202190 groups cultured in the electromagnetic field after PD98059 or SB202190 was added, and those in the control group were cultured normally.Different groups of cells were exposed to electromagnetic fields (15 Hz, 1 mT, sine wave form) for different exposure duration.The activated phosphorylated and non-phosphorylated p38MAPK were measured using Western blotting analysis with their specific corresponding antibodies.The alkaline phosphatase (ALP) activity in cells in different groups was detected according to the instructions of ALP kit.MTT assay was applied to investigate the proliferation of cells.Results Electromagnetic fields could rapidly induce the activation of p38 MAPK (P < 0.05) and the phosphorylation of p38 MAPK elevated after 15 min exposure to EMF.The phosphorylation of p38 MAPK was significantly lower in the EMF + SB202190 group than that of the EMF group.After 5 days of EMF exposure, the ALP activity of cells was significantly improved, and the effect could be inhibited by SB202190.The bone marrow mesenchymal stem cells proliferation increased significantly after being exposed to EMF for 3 days, and it could not be blocked by SB202190.Phosphorylation of ERK and MAPK increased significantly when the p38 MAPK pathway was blocked by SB202190 and exposed to EMF for 5 minutes, and it also increased significantly when the ERK MAPK pathway was blocked by PD98059 and

  2. Epithelial-Mesenchymal Transition and Breast Cancer

    Directory of Open Access Journals (Sweden)

    Yanyuan Wu

    2016-01-01

    Full Text Available Breast cancer is the most common cancer in women and distant site metastasis is the main cause of death in breast cancer patients. There is increasing evidence supporting the role of epithelial-mesenchymal transition (EMT in tumor cell progression, invasion, and metastasis. During the process of EMT, epithelial cancer cells acquire molecular alternations that facilitate the loss of epithelial features and gain of mesenchymal phenotype. Such transformation promotes cancer cell migration and invasion. Moreover, emerging evidence suggests that EMT is associated with the increased enrichment of cancer stem-like cells (CSCs and these CSCs display mesenchymal characteristics that are resistant to chemotherapy and target therapy. However, the clinical relevance of EMT in human cancer is still under debate. This review will provide an overview of current evidence of EMT from studies using clinical human breast cancer tissues and its associated challenges.

  3. Mesenchymal Stem Cells and Tooth Engineering

    Institute of Scientific and Technical Information of China (English)

    Li Peng; Ling Ye; Xue-dong Zhou

    2009-01-01

    Tooth loss compromises human oral health. Although several prosthetic methods, such as artificial denture and dental implants, are clinical therapies to tooth loss problems, they are thought to have safety and usage time issues. Recently, tooth tissue engineering has attracted more and more attention. Stem cell based tissue engineering is thought to be a promising way to replace the missing tooth. Mesenchymal stem cells (MSCs) are multipotent stem cells which can differentiate into a variety of cell types. The potential MSCs for tooth regeneration mainly include stem cells from human exfoliated deciduous teeth (SHEDs), adult dental pulp stem cells (DPSCs), stem cells from the apical part of the papilla (SCAPs), stem cells from the dental follicle (DFSCs), periodontal ligament stem cells (PDLSCs) and bone marrow derived mesenchymal stem cells (BMSCs). This review outlines the recent progress in the mesenchymal stem cells used in tooth regeneration.

  4. A Modular Approach to Triazole-Containing Chemical Inducers of Dimerisation for Yeast Three-Hybrid Screening

    Directory of Open Access Journals (Sweden)

    Nicholas J. Westwood

    2013-09-01

    Full Text Available The yeast three-hybrid (Y3H approach shows considerable promise for the unbiased identification of novel small molecule-protein interactions. In recent years, it has been successfully used to link a number of bioactive molecules to novel protein binding partners. However despite its potential importance as a protein target identification method, the Y3H technique has not yet been widely adopted, in part due to the challenges associated with the synthesis of the complex chemical inducers of dimerisation (CIDs. The development of a modular approach using potentially “off the shelf” synthetic components was achieved and allowed the synthesis of a family of four triazole-containing CIDs, MTX-Cmpd2.2-2.5. These CIDs were then compared using the Y3H approach with three of them giving a strong positive interaction with a known target of compound 2, TgCDPK1. These results showed that the modular nature of our synthetic strategy may help to overcome the challenges currently encountered with CID synthesis and should contribute to the Y3H approach reaching its full potential as an unbiased target identification strategy.

  5. Impaired skin regeneration and remodeling after cutaneous injury and chemically induced hyperplasia in taps-transgenic mice.

    Science.gov (United States)

    Hildenbrand, Maike; Rhiemeier, Verena; Hartenstein, Bettina; Lahrmann, Bernd; Grabe, Niels; Angel, Peter; Hess, Jochen

    2010-07-01

    Recently, we identified an AP-1-dependent target gene in 12-O-tetradecanoylphorbol-13-acetate (TPA)-treated mouse back skin, which encodes a retroviral-like aspartic proteinase (Taps/Asprv1). Taps expression was detected almost exclusively in stratified epithelia of mouse embryos and adult tissues, and enhanced protein levels were present in several non-neoplastic human skin disorders, implicating a crucial role for differentiation and homeostasis of multilayered epithelia. Here, we generated a mouse model in which Taps transgene expression is under the control of the human ubiquitin C promoter (UBC-Taps). Although no obvious phenotype was observed in normal skin development and homeostasis, these mice showed a significant delay in cutaneous wound closure compared with control animals. Shortly after re-epithelialization, we found an increase in keratinocytes in the stratum granulosum, which express Filaggrin, a late differentiation marker. A hypergranulosum-like phenotype with increased numbers of Filaggrin-positive keratinocytes was also observed in UBC-Taps mice after administration of TPA. In summary, these data show that aberrant Taps expression causes impaired skin regeneration and skin remodeling after cutaneous injury and chemically induced hyperplasia. PMID:20237492

  6. Demonstration of feasibility of in vivo gene therapy for Gaucher disease using a chemically induced mouse model.

    Science.gov (United States)

    Marshall, John; McEachern, Kerry Anne; Kyros, Julie A Cavanagh; Nietupski, Jennifer B; Budzinski, Tracey; Ziegler, Robin J; Yew, Nelson S; Sullivan, Jennifer; Scaria, Abraham; van Rooijen, Nico; Barranger, John A; Cheng, Seng H

    2002-08-01

    Progress towards developing gene therapy for Gaucher disease has been hindered by the lack of an animal model. Here we describe a mouse model of Gaucher disease which has a chemically induced deficiency of glucocerebrosidase and that accumulates elevated levels of glucosylceramide (GL-1) in the lysosomes of Kupffer cells. Administration of mannose-terminated glucocerebrosidase (Cerezyme) resulted in the reduction of GL-1 levels in the livers of these animals. Gene transduction of hepatocytes with a plasmid DNA vector encoding human glucocerebrosidase (pGZB-GC) generated high-level expression and secretion of the enzyme into systemic circulation with consequent normalization of Kupffer cell GL-1 levels. This suggested that the de novo synthesized and unmodified enzyme produced by hepatocyte transduction was also capable of being delivered to the cells that are primarily affected in Gaucher disease. Immunolocalization studies also revealed that preferential transduction and expression of human glucocerebrosidase in the Kupffer cells with subsequent reduction in the GL-1 levels could be attained with a low dose of a recombinant adenoviral vector encoding the human enzyme (Ad2/CMV-GC). This observation raises the possibility of gene therapy for Gaucher disease that involves directly transducing the affected histiocytes using recombinant adenoviral vectors. Together, these data demonstrate the potential for use of in vivo gene therapy vectors for treating Gaucher disease. PMID:12161184

  7. Dormancy in potato tuber meristems: chemically induced cessation in dormancy matches the natural process based on transcript profiles.

    Science.gov (United States)

    Campbell, Michael; Segear, Erika; Beers, Lee; Knauber, Donna; Suttle, Jeffrey

    2008-11-01

    Meristem dormancy in perennial plants is a developmental process that results in repression of metabolism and growth. The cessation of dormancy results in rapid growth and should be associated with the production of nascent transcripts that encode for gene products controlling for cell division and growth. Dormancy cessation was allowed to progress normally or was chemically induced using bromoethane (BE), and microarray analysis was used to demonstrate changes in specific transcripts in response to dormancy cessation before a significant increase in cell division. Comparison of normal dormancy cessation to BE-induced dormancy cessation revealed a commonality in both up and downregulated transcripts. Many transcripts that decrease as dormancy terminates are inducible by abscisic acid particularly in the conserved BURP domain proteins, which include the RD22 class of proteins and in the storage protein patatin. Transcripts that are associated with an increase in expression encoded for proteins in the oxoglutarate-dependent oxygenase family. We conclude that BE-induced cessation of dormancy initiates transcript profiles similar to the natural processes that control dormancy. PMID:18317824

  8. Matrix metalloproteinases regulate the formation of dendritic spine head protrusions during chemically induced long-term potentiation.

    Directory of Open Access Journals (Sweden)

    Zsuzsanna Szepesi

    Full Text Available Dendritic spines are are small membranous protrusions that extend from neuronal dendrites and harbor the majority of excitatory synapses. Increasing evidence has shown that matrix metalloproteinases (MMPs, a family of extracellularly acting and Zn(2+-dependent endopeptidases, are able to rapidly modulate dendritic spine morphology. Spine head protrusions (SHPs are filopodia-like processes that extend from the dendritic spine head, representing a form of postsynaptic structural remodeling in response to altered neuronal activity. Herein, we show that chemically induced long-term potentiation (cLTP in dissociated hippocampal cultures upregulates MMP-9 activity that controls the formation of SHPs. Blocking of MMPs activity or microtubule dynamics abolishes the emergence of SHPs. In addition, autoactive recombinant MMP-9, promotes the formation of SHPs in organotypic hippocampal slices. Furthermore, spines with SHPs gained postsynaptic α-amino-3-hydroxyl-5-methyl-4-isoxazole propionic acid (AMPA receptors upon cLTP and the synaptic delivery of AMPA receptors was controlled by MMPs. The present results strongly imply that MMP-9 is functionally involved in the formation of SHPs and the control of postsynaptic receptor distribution upon cLTP.

  9. Mesenchymal Stem Cells (MSC) Prevented the Progression of Renovascular Hypertension, Improved Renal Function and Architecture

    OpenAIRE

    Oliveira-Sales, Elizabeth B.; MAQUIGUSSA, EDGAR; Semedo, Patricia; PEREIRA, LUCIANA G; Ferreira, Vanessa M; Câmara, Niels O.; Bergamaschi, Cassia T.; Campos, Ruy R.; Mirian A. Boim

    2013-01-01

    Renovascular hypertension induced by 2 Kidney-1 Clip (2K-1C) is a renin-angiotensin-system (RAS)-dependent model, leading to renal vascular rarefaction and renal failure. RAS inhibitors are not able to reduce arterial pressure (AP) and/or preserve the renal function, and thus, alternative therapies are needed. Three weeks after left renal artery occlusion, fluorescently tagged mesenchymal stem cells (MSC) (2×105 cells/animal) were injected weekly into the tail vein in 2K-1C hypertensive rats....

  10. Sprague-Dawley大鼠外周血来源间充质干细胞的动员、分离培养及鉴定%Mobilization, Culture,Isolation and Identification of Mesenchymal Stem Cells from Peripheral Blood of Sprague-Dawley Rats

    Institute of Scientific and Technical Information of China (English)

    张继英; 刘刚; 马栋; 付维力; 余家阔

    2012-01-01

    Objective The purpose of this study was to explore the methods of mobilization, isolation, culture and identification of mesenchymal stem cells (MSC) in rat peripheral blood,in order to find a new source for seeding cells with minimally invasion for cell therapy and tissue engineering products culturing in the field of sports traumatology. Methods MSC in peripheral blood were mobilized by administration of combined drugs. Mononuclear cells in blood samples from the abdominal aorta were isolated by lymphocyte separation medium and density gradient centrifugation, then cultured in vitro to the third passage through microscopic observation of morphological characteristics and growth state. The phenotype of the third passage MSC was identified, and the differentiation capacity into bone,adipose and cartilage was confirmed. Results After 3-5 days culture in vitro,a few isolated cells in spindle shape adhered to the plastic walls. And after 7-9 days culture in vitro,cells cluster and colonies appeared. The third passage cells showed uniform morphology and good proliferation with expression of the typical surface markers of MSC,including CD44 and CD90,and without expression of surface markers of hematopoietic stem cells such as CD34 and CD45. After 21-day induction of bone formation,the cells demonstrated positive staining for Alizarin red and ALP,and formation of mineralization nodes. Positive staining for Oil red 0 and for Tolridine blue, and immunohistochemical staining for type II collagen confirmed the formation of cartilage-specific extracellular aggrecans and type II collagen. Conclusion These methods for mobilization by administration of combined drugs could increase the release of stem cells from peripheral blood. MSC cultured in vitro had multipotent capacity of differentiation along three lineages. MSC derived from peripheral blood could be a potential source of seeding cells for cell therapy and tissue engineering.%目的:探讨Sprague -Dawley大鼠外周血来源间充质干细胞(mesenchymal

  11. Epithelial-Mesenchymal Transition in Pancreatic Carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Maier, Harald J., E-mail: harald.maier@uni-ulm.de; Wirth, Thomas [Institute of Physiological Chemistry, University of Ulm, 89081 Ulm (Germany); Beug, Hartmut [Institute of Molecular Pathology, 1030 Vienna (Austria)

    2010-12-09

    Pancreatic carcinoma is the fourth-leading cause of cancer death and is characterized by early invasion and metastasis. The developmental program of epithelial-mesenchymal transition (EMT) is of potential importance for this rapid tumor progression. During EMT, tumor cells lose their epithelial characteristics and gain properties of mesenchymal cells, such as enhanced motility and invasive features. This review will discuss recent findings pertinent to EMT in pancreatic carcinoma. Evidence for and molecular characteristics of EMT in pancreatic carcinoma will be outlined, as well as the connection of EMT to related topics, e.g., cancer stem cells and drug resistance.

  12. Epithelial-Mesenchymal Transition in Pancreatic Carcinoma

    International Nuclear Information System (INIS)

    Pancreatic carcinoma is the fourth-leading cause of cancer death and is characterized by early invasion and metastasis. The developmental program of epithelial-mesenchymal transition (EMT) is of potential importance for this rapid tumor progression. During EMT, tumor cells lose their epithelial characteristics and gain properties of mesenchymal cells, such as enhanced motility and invasive features. This review will discuss recent findings pertinent to EMT in pancreatic carcinoma. Evidence for and molecular characteristics of EMT in pancreatic carcinoma will be outlined, as well as the connection of EMT to related topics, e.g., cancer stem cells and drug resistance

  13. Epithelial-Mesenchymal Transition in Pancreatic Carcinoma

    Directory of Open Access Journals (Sweden)

    Thomas Wirth

    2010-12-01

    Full Text Available Pancreatic carcinoma is the fourth-leading cause of cancer death and is characterized by early invasion and metastasis. The developmental program of epithelial-mesenchymal transition (EMT is of potential importance for this rapid tumor progression. During EMT, tumor cells lose their epithelial characteristics and gain properties of mesenchymal cells, such as enhanced motility and invasive features. This review will discuss recent findings pertinent to EMT in pancreatic carcinoma. Evidence for and molecular characteristics of EMT in pancreatic carcinoma will be outlined, as well as the connection of EMT to related topics, e.g., cancer stem cells and drug resistance.

  14. 宫内移植骨髓间充质干细胞对显性脊柱裂胎鼠神经元重建的作用研究%Neuron regeneration of in utero mesenchymal stem cell transplantation in rat fetuses with spina bifida aperta

    Institute of Scientific and Technical Information of China (English)

    马丽丽; 袁正伟; 蒋俊红; 陈海琛; 杨兆辉; 姜艳; 徐延波

    2014-01-01

    目的 胚胎期移植骨髓间充质干细胞(bone marrow stem cell,BMSCs)至先天性脊柱裂模型胎鼠脊髓中,观察其在显性脊柱裂胎鼠脊髓中存活及分化情况,并探讨其对脊髓神经元修复的治疗潜能.方法 利用胎仔外科和显微注射技术将绿色荧光蛋白(eGFP)转染的BMSCs分别在胚胎第16、17和18天移植到83只脊柱裂和20只正常大鼠胚胎中(group E16、E17and E18),胚胎20 d的胎鼠脊髓被取出并固定.经冰冻切片后,计数eGFP阳性细胞数目,并利用免疫荧光方法检测脊髓中eGFP阳性的BMSCs中神经干细胞(Nestin)、神经元(Neurofilament)和神经胶质细胞(GFAP)的分布与表达.结果 本研究成功的将MSC在宫内移植入胎鼠体内,孕鼠的存活率为94%(81/86),胎鼠的存活率为78.1%(136/174);实验中孕16d的胎鼠存活率为75.8%(91/120)低于孕17 d的80.6%(25/31)和孕18d的87.0%(20/23).计数脊髓中eGFP阳性细胞数,范围在166~735个/根,其中P0代MSCs存活率最低,为10.8%;P3~6代为22.6%、P10~14代为20.1%;P10~14代MSC在母鼠孕16d时移植得到最佳分化率;统计eGFP阳性细胞表达Nestin、Neurofilament和GFAP的阳性率分别为4%~5%、1%~2%和5%~7%.结论 BMSCs移植到显性脊柱裂胎鼠脊髓中能够存活,并在脊髓中表达Nestin、Neurofilament和GFAP,提示移植骨髓间充质干细胞具有向神经干细胞、神经元和神经胶质细胞分化的可能.%Objective To observe the survival and differentiation status of transplanted bone marrow mesenchymal stem cells (BM-MSCs) in spinal cord of dominant spina bifida fetal rats and discuss their therapeutic potentials for repairing spinal cord neurons.Methods Fetal surgery and micro-injection technology were employed to transplant green fluorescent protein (eGFP)-transfected BMSCs into 83 spina bifida and 20 normal rat embryos respectively at embryonic days 16,17 and 18 (groups E16,E17 & E18).And,at Day 20,spinal cord was

  15. 不同途径骨髓间充质干细胞移植对颅脑损伤大鼠海马的作用%Effects of bone marrow mesenchymal stem cells transplantation in different ways on hippocanipal neurons following traumatic brain injury in rats

    Institute of Scientific and Technical Information of China (English)

    吕加希; 尹宏

    2012-01-01

    背景:近年来骨髓间充质干细胞移植在中枢神经系统中的应用越来越广泛,研究其最佳的移植方式具有重要意义.目的:观察不同途径移植骨髓间充质干细胞对创伤性脑损伤大鼠海马神经元的影响.方法:分离培养Wistar大鼠骨髓间充质干细胞,以PKH26标记第3代骨髓间充质干细胞,荧光显微镜观察并流式细胞仪检测标记率;采用改进Feeney法制作脑损伤致海马神经元损伤模型.60只Wistar大鼠随机分为3组,脑损伤区移植组:造模后注射含骨髓间充质干细胞的生理盐水10 μL;尾静脉移植组:自尾静脉注入含骨髓间充质干细胞的生理盐水1 mL;创伤性脑损伤组:不给予任何处理.结果与结论:荧光显微镜观察PKH26标记的BMSCs呈球形,呈现均匀分布的红色荧光;PKH26标记BMSCs的阳性率>99%.脑损伤区移植组大鼠学习记忆功能明显好于其他两组,PKH26阳性细胞和苏木精-伊红染色切片中的神经元数量脑损伤区移植组多于尾静脉移植组,尾静脉移植组多于创伤性脑损伤组(P < 0.05).GAP-43 mRNA的表达脑损伤区移植组高于尾静脉移植组,尾静脉移植组高于创伤性脑损伤组(P < 0.05).提示移植骨髓间充质干细胞对创伤性脑损伤大鼠海马神经元有保护作用,脑损伤区移植优于尾静脉移植.%BACKGROUND: In recent years, applicati on of bone marrow mesenchymal stem cells (BMSCs) transplantation in the central nervous system has become more widely, and the study of the best way to transplantation is important. OBJECTIVE: To investigate the effect of BMSCs transplantation in different ways on hippocanipal neuron following traumatic brain injury (TBI) in rats.METHODS: BMSCs were isolated from the bone marrow of 1-month-old SD rat. The BMSCs at passage 3 were labeled with PKH26 and were observed under fluorescence microscope. The percentage of the labeled cells was detected byflowcytometer. The model of hippocampal neuronal death

  16. Supernatant of Bone Marrow Mesenchymal Stromal Cells Induces Peripheral Blood Mononuclear Cells Possessing Mesenchymal Features

    OpenAIRE

    Hu, Gang; Xu, Jun-jun; Deng, Zhi-Hong; Feng, Jie; Jin, Yan

    2011-01-01

    Increasing evidence shows that some cells from peripheral blood fibroblast-like mononuclear cells have the capacity to differentiate into mesenchymal lineages. However, the insufficiency of these cells in the circulation challenges the cell isolation and subsequently limits the clinical application of these cells. In the present study, the peripheral blood mononuclear cells (pbMNCs) were isolated from wound animals and treated with the supernatant of bone marrow mesenchymal stromal cells (bmM...

  17. Neural differentiation of human placenta-derived mesenchymal stem cells following neural cell co-culture

    Institute of Scientific and Technical Information of China (English)

    Nailong Yang; Hongyan Zhang; Xiaojuan Sun; Lili Xu

    2011-01-01

    We induced human placenta-derived mesenchymal stem cells (hPMSCs) to differentiate into neural cells by adding chemical reagents,despite the fact that toxic chemicals induce cell shrinkage or cytoskeletal formation,which does not represent a proper cell differentiation process.The present study established a co-culture system with hPMSCs and neural cells and analyzed the influence of neural cells on hPMSC differentiation in a co-culture system.hPMSCs were isolated and purified from human full-term placenta using collagenase digestion.Fetal neural cells were co-cultured with hPMSCs for 48 hours using the Transwell co-culture system.hPMSCs co-cultured with neural cells exhibited a slender morphology with a filament.After 96 hours,hPMSCs expressed neuron-specific enolase,which suggested that co-culture of hPMSCs and neural cells induced neural differentiation of hPMSCs.

  18. Cryopreservation and revival of mesenchymal stromal cells

    DEFF Research Database (Denmark)

    Haack-Sørensen, Mandana; Kastrup, Jens

    2011-01-01

    initiated. As there has been a precedent for the use of bone marrow stem cells in the treatment of hematological malignancies and ischemic heart diseases through randomized clinical safety and efficacy trials, the development of new therapies based on culture-expanded human mesenchymal stromal cells (MSCs...

  19. Mesenchymal stem cells in oral reconstructive surgery

    DEFF Research Database (Denmark)

    Jakobsen, C; Sørensen, J A; Kassem, M;

    2013-01-01

    This study evaluated clinical outcomes following intraoperative use of adult mesenchymal stem cells (MSCs) in various oral reconstructive procedures. PubMed was searched without language restrictions from 2000 to 2011 using the search words stem cell, oral surgery, tissue engineering, sinus lift...

  20. Effects of mesenchymal stem cells transplantation on ischemia-reperfusion injury of intestine in rats%骨髓间充质干细胞移植对大鼠小肠缺血再灌注损伤的影响

    Institute of Scientific and Technical Information of China (English)

    张静; 沈中阳; 宋红丽; 董冲; 郑卫萍; 郭瑞雪; 杜杰静

    2012-01-01

    目的 探讨大鼠骨髓间充质干细胞( BMSC)移植对其小肠缺血再灌注损伤的影响.方法 获取健康Wistar大鼠骨髓,体外提取、培养BMSC,收获传代培养的第3代细胞并鉴定其表型.在健康Wistar大鼠肠黏膜下注射皮肤黑色素瘤细胞,并用底物荧光素法示踪细胞.另外制作Wistar大鼠肠道缺血再灌注模型,将模型大鼠分为2组:实验组大鼠在肠黏膜下注射BMSC悬液1 ml(含5×106个细胞);对照组大鼠在肠黏膜下注射等量生理盐水.分别于术后0、2、6、24、72和120h处死大鼠,留取血清和小肠组织样本.采用酶联免疫吸附试验检测血清二胺氧化酶( DAO)、D-乳酸和肿瘤坏死因子α(TNF-α),用光镜和透射电镜观察肠组织,蛋白质印迹法和免疫组织化学法检测紧密连接蛋白-1(ZO-1).结果 体外成功分离培养出BMSC.经肠黏膜移植后的皮肤黑色素瘤细胞定植在肠道.实验组大鼠小肠病理改变较对照组轻,小肠黏膜屏障更完好.术后6h实验组DAO为(11.36±1.89) IU/ml,对照组为(14.27±2.09) IU/ml(P<0.05);24 h时实验组DAO为(5.04±1.04)IU/ml,对照组为(7.35±1.46) IU/ml(P<0.05).术后6 h实验组D-乳酸为(1.57±0.25) mmol/L,对照组为(1.93±0.19)mmol/L(P<0.05);术后24 h实验组D-乳酸为(1.09±0.13)mmol/L,对照组为(1.41±0.07) mmol/L(P<0.01).术后6h实验组TNF-α为(266.09±8.84)ng/L,对照组为(286.81±11.54) ng/L (P<0.01);术后24 h实验组TNF-α为(190.39±4.24) ng/L,对照组为(218.49±15.51 )ng/L(P<0.01).实验组ZO-1的表达高于对照组.结论 肠黏膜下移植BMSC可以减轻小肠缺血再灌注损伤.%Objective To study the effects of bone marrow mesenchymal stem cells (BMSCs) transplantation on the ischemia-reperfusion injury of the intestine in rats.Methods BMSCs were isolated from femur of male Wistar rats and cultured,and the phenotypes of third generation cultured cells were identified.B16-F10-Luc-G5 cells were injected into the

  1. Sieve-based coreference resolution enhances semi-supervised learning model for chemical-induced disease relation extraction

    Science.gov (United States)

    Le, Hoang-Quynh; Tran, Mai-Vu; Dang, Thanh Hai; Ha, Quang-Thuy; Collier, Nigel

    2016-01-01

    The BioCreative V chemical-disease relation (CDR) track was proposed to accelerate the progress of text mining in facilitating integrative understanding of chemicals, diseases and their relations. In this article, we describe an extension of our system (namely UET-CAM) that participated in the BioCreative V CDR. The original UET-CAM system’s performance was ranked fourth among 18 participating systems by the BioCreative CDR track committee. In the Disease Named Entity Recognition and Normalization (DNER) phase, our system employed joint inference (decoding) with a perceptron-based named entity recognizer (NER) and a back-off model with Semantic Supervised Indexing and Skip-gram for named entity normalization. In the chemical-induced disease (CID) relation extraction phase, we proposed a pipeline that includes a coreference resolution module and a Support Vector Machine relation extraction model. The former module utilized a multi-pass sieve to extend entity recall. In this article, the UET-CAM system was improved by adding a ‘silver’ CID corpus to train the prediction model. This silver standard corpus of more than 50 thousand sentences was automatically built based on the Comparative Toxicogenomics Database (CTD) database. We evaluated our method on the CDR test set. Results showed that our system could reach the state of the art performance with F1 of 82.44 for the DNER task and 58.90 for the CID task. Analysis demonstrated substantial benefits of both the multi-pass sieve coreference resolution method (F1 + 4.13%) and the silver CID corpus (F1 +7.3%). Database URL: SilverCID–The silver-standard corpus for CID relation extraction is freely online available at: https://zenodo.org/record/34530 (doi:10.5281/zenodo.34530).

  2. Paramagnetic particles carried by cell-penetrating peptide tracking of bone marrow mesenchymal stem cells, a research in vitro

    International Nuclear Information System (INIS)

    The ability to track the distribution and differentiation of stem cells by high-resolution imaging techniques would have significant clinical and research implications. In this study, a model cell-penetrating peptide was used to carry gadolinium particles for magnetic resonance imaging of the mesenchymal stem cells. The mesenchymal stem cells were isolated from rat bone marrow by Percoll and identified by osteogenic differentiation in vitro. The cell-penetrating peptides labeled with fluorescein-5-isothiocyanate and gadolinium were synthesized by a solid-phase peptide synthesis method and the relaxivity of cell-penetrating peptide-gadolinium paramagnetic conjugate on 400 MHz nuclear magnetic resonance was 5.7311 ± 0.0122 mmol-1 s-1, higher than that of diethylenetriamine pentaacetic acid gadolinium (p < 0.05). Fluorescein imaging confirmed that this new peptide could internalize into the cytoplasm and nucleus. Gadolinium was efficiently internalized into mesenchymal stem cells by the peptide in a time- or concentration-dependent fashion, resulting in intercellular T1 relaxation enhancement, which was obviously detected by 1.5 T magnetic resonance imaging. Cytotoxicity assay and flow cytometric analysis showed the intercellular contrast medium incorporation did not affect cell viability and membrane potential gradient. The research in vitro suggests that the newly constructed peptides could be a vector for tracking mesenchymal stem cells

  3. Application of human amniotic mesenchymal cells as an allogeneic transplantation cell source in bone regenerative therapy

    International Nuclear Information System (INIS)

    Autogenous mesenchymal stem cells (MSCs) have therapeutic applications in bone regenerative therapy due to their pluripotency. However, the ability of MSCs to proliferate and differentiate varies between donors. Furthermore, alternative sources of MSCs are required for patients with contraindications to autogenous cell therapy. The aim of this study was to evaluate the potential of mesenchymal cells from the human amniotic membrane (HAM) as a source of cells for allogeneic transplantation in bone regenerative therapy. Cells that retained a proliferative capacity of more than 50 population doubling level were distinguished from other HAM cells as HAMα cells and induced to osteogenic status—their in vivo osteogenesis was subsequently investigated in rats. It was found that HAMα cells were spindle shaped and were positive for MSC markers and negative for hematopoietic stem cell markers. Alkaline phosphatase activity and calcium deposition increased with osteogenic status of HAMα cells. The expression of osteocalcin mRNA was increased in HAMα cells cultured on calcium phosphate scaffolds. Moreover, xenografted HAMα cells remained viable and produced extracellular matrix for several weeks. Thus, this study suggests that human amniotic mesenchymal cells possess osteogenic differentiation potential and could be applied to allogeneic transplantation in bone regenerative therapy. - Highlights: ► Human amniotic mesenchymal cells include cells (HAMα cells) that have the properties of MSCs. ► HAMα cells have excellent osteogenic differentiation potential. ► Osteogenic differentiation ability of HAMα was amplified by calcium phosphate scaffolds. ► HAMα cells can be applicable to allogeneic cell transplantation in bone regenerative therapy.

  4. Application of human amniotic mesenchymal cells as an allogeneic transplantation cell source in bone regenerative therapy

    Energy Technology Data Exchange (ETDEWEB)

    Tsuno, Hiroaki [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Department of Oral and Maxillofacial Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Yoshida, Toshiko [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Nogami, Makiko [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Department of Orthopedic Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Koike, Chika; Okabe, Motonori [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Noto, Zenko [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Department of Oral and Maxillofacial Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Arai, Naoya; Noguchi, Makoto [Department of Oral and Maxillofacial Surgery, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan); Nikaido, Toshio, E-mail: tnikaido@med.u-toyama.ac.jp [Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani Toyama, Toyama 930-0194 (Japan)

    2012-12-01

    Autogenous mesenchymal stem cells (MSCs) have therapeutic applications in bone regenerative therapy due to their pluripotency. However, the ability of MSCs to proliferate and differentiate varies between donors. Furthermore, alternative sources of MSCs are required for patients with contraindications to autogenous cell therapy. The aim of this study was to evaluate the potential of mesenchymal cells from the human amniotic membrane (HAM) as a source of cells for allogeneic transplantation in bone regenerative therapy. Cells that retained a proliferative capacity of more than 50 population doubling level were distinguished from other HAM cells as HAM{alpha} cells and induced to osteogenic status-their in vivo osteogenesis was subsequently investigated in rats. It was found that HAM{alpha} cells were spindle shaped and were positive for MSC markers and negative for hematopoietic stem cell markers. Alkaline phosphatase activity and calcium deposition increased with osteogenic status of HAM{alpha} cells. The expression of osteocalcin mRNA was increased in HAM{alpha} cells cultured on calcium phosphate scaffolds. Moreover, xenografted HAM{alpha} cells remained viable and produced extracellular matrix for several weeks. Thus, this study suggests that human amniotic mesenchymal cells possess osteogenic differentiation potential and could be applied to allogeneic transplantation in bone regenerative therapy. - Highlights: Black-Right-Pointing-Pointer Human amniotic mesenchymal cells include cells (HAM{alpha} cells) that have the properties of MSCs. Black-Right-Pointing-Pointer HAM{alpha} cells have excellent osteogenic differentiation potential. Black-Right-Pointing-Pointer Osteogenic differentiation ability of HAM{alpha} was amplified by calcium phosphate scaffolds. Black-Right-Pointing-Pointer HAM{alpha} cells can be applicable to allogeneic cell transplantation in bone regenerative therapy.

  5. The effect of phospholipid transfer protein on cigarette smoke extract induced epithelial-mesenchymal transition of rat alveolar type Ⅱ cells%磷脂转运蛋白在烟草诱导RLE-6TN细胞株发生上皮间质转化中的作用

    Institute of Scientific and Technical Information of China (English)

    巫凤苹; 陈亚娟; 余秀英; 廖科; 李丹丹; 陈虹

    2016-01-01

    Objective To investigate the effect of phospholipid transfer protein(PLTP) on cigarette smoke extract(CSE) induced epithelial-mesenchymal transition(EMT) in rat alveolar Type Ⅱ cells (RLE-6TN).Methods CSE of different concentrations (0%,0.25%,0.5% and 1%) was co-cultured for 2 or 3days with RLE-6TN,either pre-treated or not pre-treated with siRNA-PLTP for 6 h.Expression levels of E-cadherin mRNA and Vimentin mRNA were examined by RT-PCR,while expression levels of PLTP,E-cadherin,N-cadherin and Vimentin were examined by Western blot.Results Our results showed that the expression of E-cadherin mRNA decreased in CSE-treated groups:1.01 ± 0.05,0.74 ± 0.05,0.65 ± 0.03,0.30 ±0.08 respectively at different concentrations of CSE (0 %,0.25%,0.5 %,and 1.0%);while the level of Vimentin mRNA increased significantly in 1% CSE treated cells (1.88 ± O.49),compared with control cells (1.01 ±0.20).Treatment with CSE at different concentrations (0%,0.25%,0.5% and 1%) showed that the protein levels of PLTP were 0.42 ± 0.02,0.89 ± 0.25,1.08 ± 0.18,1.61 ± 0.06 respectively;those of E-cadherin were 1.61 ± 0.04,1.08 ± 0.10,0.62 ± 0.08,0.68 ± 0.17,respectively;those of N-cadherin were 0.60 ± 0.14,0.57 ± 0.26,0.88 ± 0.30,1.94 ± 0.54,respectively;and those of Vimentin were 0.61 ± 0.05,0.98 ± 0.16,1.07 ± 0.14,1.34 ± 0.19,respectively;all P < 0.05 when the 1% CSE group was compared with the control group.EMT induced by CSE was significantly inhibited by siRNA-PLTP.Conclusion PLTP may be involved in CSE induced EMT of rat alveolar cells.%目的 探讨磷脂转运蛋白(PLTP)在烟草提取物(CSE)诱导大鼠Ⅱ型肺泡上皮细胞株RLE-6TN发生上皮间质转化(EMT)中的作用.方法 体外培养RLE-6TN细胞株24 h,分为4组,每组3孔,分别加入0%、0.25%、0.5%和1% CSE培养2d,检测E-钙黏蛋白和波形蛋白mRNA表达以及细胞和CSE共培养3d检测PLTP、EMT相关蛋白(E-钙黏蛋白、N-钙黏蛋白和波形

  6. 大鼠骨髓间质干细胞的BrdU体外标记和体内示踪研究%In vitro Analysis and in vivo Tracing of BrdU-labeled Rat Bone Marrow Mesenchymal Stem Cells

    Institute of Scientific and Technical Information of China (English)

    张明鸣; 贾贵清; 程惊秋; 杨平; 陆燕蓉; 伍晓汀

    2012-01-01

    Objectives To investigate the optimal condition of bromodeoxyuridine (BrdU) labeling for bone marrow mesenchymal stem cells (BMSCs) in vitro and the feasibility of in vivo tracing of BrdU-labeling BMSCs. Methods BMSCs were isolated from Wistar rats and were in vitro routinely cultured. The third passage BMSCs was used for identification of special surface antigens by immunohistochemical methods. The purified BMSCs were incubated with BrdU at different concentrations for different incubating time to investigate optimal BrdU concentration and incubating time for cell labeling. The cell labeling index of BrdU was calculated with immunohistochemical analysis. BMSCs labeled BrdU were injected into damaged gastric mucosa of rats by micro injector. The colonization of BMSCs labeled BrdU in gastric mucosa was viewed. Results After purification and proliferation, the primary cultured BMSCs were uniformly long spindle-shapped form and formed cell colony, which showed the characteristics of stem cell. Immunocytochemistry showed BMSCs were positive for CD44 and CD90, while negative for CD14, CD45. The labeling rate of BrdU increased with the labeling time lasting and reached its height at 48 h. After incubating 48 and 72 hours, the labeling rate of BrdU with a concentration of 10 μmol/L was higher than that of BrdU with a concentration of 5 μmol/L (P 0. 05). In addition, the BrdU labeling could be detected after five consecutive passages and the labeling time could keep 21 d. The pathological observation demonstrated that BrdU-labeled BMSCs could colonize the damaged gastric mucosa with normal morphologic characteristics during observation period. Conclusion BrdU labeling might be a feasible method for dynamic observation of the migration, growth and differentiation of migrating BMSCs in colonizing sites.%目的 探讨骨髓间质干细