WorldWideScience

Sample records for bovine ivf embryos

  1. Who abandons embryos after IVF?

    LENUS (Irish Health Repository)

    Walsh, A P H

    2010-04-01

    This investigation describes features of in vitro fertilisation (IVF) patients who never returned to claim their embryos following cryopreservation. Frozen embryo data were reviewed to establish communication patterns between patient and clinic; embryos were considered abandoned when 1) an IVF patient with frozen embryo\\/s stored at our facility failed to make contact with our clinic for > 2 yrs and 2) the patient could not be located after a multi-modal outreach effort was undertaken. For these patients, telephone numbers had been disconnected and no forwarding address was available. Patient, spouse and emergency family contact\\/s all escaped detection efforts despite an exhaustive public database search including death records and Internet directory portals. From 3244 IVF cycles completed from 2000 to 2008, > or = 1 embryo was frozen in 1159 cases (35.7%). Those without correspondence for > 2 yrs accounted for 292 (25.2%) patients with frozen embryos; 281 were contacted by methods including registered (signature involving abandoned embryos did not differ substantially from other patients. The goal of having a baby was achieved by 10\\/11 patients either by spontaneous conception, adoption or IVF. One patient moved away with conception status unconfirmed. The overall rate of embryo abandonment was 11\\/1159 (< 1%) in this IVF population. Pre-IVF counselling minimises, but does not totally eliminate, the problem of abandoned embryos. As the number of abandoned embryos from IVF accumulates, their fate urgently requires clarification. We propose that clinicians develop a policy consistent with relevant Irish Constitutional provisions to address this medical dilemma.

  2. Efficient edition of the bovine PRNP prion gene in somatic cells and IVF embryos using the CRISPR/Cas9 system.

    Science.gov (United States)

    Bevacqua, R J; Fernandez-Martín, R; Savy, V; Canel, N G; Gismondi, M I; Kues, W A; Carlson, D F; Fahrenkrug, S C; Niemann, H; Taboga, O A; Ferraris, S; Salamone, D F

    2016-11-01

    The recently developed engineered nucleases, such as zinc-finger nucleases, transcription activator-like effector nucleases, and clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease (Cas) 9, provide new opportunities for gene editing in a straightforward manner. However, few reports are available regarding CRISPR application and efficiency in cattle. Here, the CRISPR/Cas9 system was used with the aim of inducing knockout and knock-in alleles of the bovine PRNP gene, responsible for mad cow disease, both in bovine fetal fibroblasts and in IVF embryos. Five single-guide RNAs were designed to target 875 bp of PRNP exon 3, and all five were codelivered with Cas9. The feasibility of inducing homologous recombination (HR) was evaluated with a reporter vector carrying EGFP flanked by 1 kbp PRNP regions (pHRegfp). For somatic cells, plasmids coding for Cas9 and for each of the five single-guide RNAs (pCMVCas9 and pSPgRNAs) were transfected under two different conditions (1X and 2X). For IVF zygotes, cytoplasmic injection was conducted with either plasmids or mRNA. For plasmid injection groups, 1 pg pCMVCas9 + 0.1 pg of each pSPgRNA (DNA2X) was used per zygote. In the case of RNA, two amounts (RNA1X and RNA2X) were compared. To assess the occurrence of HR, a group additionally cotransfected or coinjected with pHRegfp plasmid was included. Somatic cell lysates were analyzed by polymerase chain reaction and surveyor assay. In the case of embryos, the in vitro development and the genotype of blastocysts were evaluated by polymerase chain reaction and sequencing. In somatic cells, 2X transfection resulted in indels and large deletions of the targeted PRNP region. Regarding embryo injection, higher blastocyst rates were obtained for RNA injected groups (46/103 [44.6%] and 55/116 [47.4%] for RNA1X and RNA2X) than for the DNA2X group (26/140 [18.6%], P genetic modifications (29) was higher than the total number of gene-edited embryos, as

  3. Efficient edition of the bovine PRNP prion gene in somatic cells and IVF embryos using the CRISPR/Cas9 system.

    Science.gov (United States)

    Bevacqua, R J; Fernandez-Martín, R; Savy, V; Canel, N G; Gismondi, M I; Kues, W A; Carlson, D F; Fahrenkrug, S C; Niemann, H; Taboga, O A; Ferraris, S; Salamone, D F

    2016-11-01

    The recently developed engineered nucleases, such as zinc-finger nucleases, transcription activator-like effector nucleases, and clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease (Cas) 9, provide new opportunities for gene editing in a straightforward manner. However, few reports are available regarding CRISPR application and efficiency in cattle. Here, the CRISPR/Cas9 system was used with the aim of inducing knockout and knock-in alleles of the bovine PRNP gene, responsible for mad cow disease, both in bovine fetal fibroblasts and in IVF embryos. Five single-guide RNAs were designed to target 875 bp of PRNP exon 3, and all five were codelivered with Cas9. The feasibility of inducing homologous recombination (HR) was evaluated with a reporter vector carrying EGFP flanked by 1 kbp PRNP regions (pHRegfp). For somatic cells, plasmids coding for Cas9 and for each of the five single-guide RNAs (pCMVCas9 and pSPgRNAs) were transfected under two different conditions (1X and 2X). For IVF zygotes, cytoplasmic injection was conducted with either plasmids or mRNA. For plasmid injection groups, 1 pg pCMVCas9 + 0.1 pg of each pSPgRNA (DNA2X) was used per zygote. In the case of RNA, two amounts (RNA1X and RNA2X) were compared. To assess the occurrence of HR, a group additionally cotransfected or coinjected with pHRegfp plasmid was included. Somatic cell lysates were analyzed by polymerase chain reaction and surveyor assay. In the case of embryos, the in vitro development and the genotype of blastocysts were evaluated by polymerase chain reaction and sequencing. In somatic cells, 2X transfection resulted in indels and large deletions of the targeted PRNP region. Regarding embryo injection, higher blastocyst rates were obtained for RNA injected groups (46/103 [44.6%] and 55/116 [47.4%] for RNA1X and RNA2X) than for the DNA2X group (26/140 [18.6%], P < 0.05). In 46% (26/56) of the total sequenced blastocysts, specific gene editing was

  4. Effects of oocyte quality, incubation time and maturation environment on the number of chromosomal abnormalities in IVF-derived early bovine embryos.

    Science.gov (United States)

    Demyda-Peyrás, Sebastian; Dorado, Jesus; Hidalgo, Manuel; Anter, Jaouad; De Luca, Leonardo; Genero, Enrique; Moreno-Millán, Miguel

    2013-01-01

    Chromosomal aberrations are one of the major causes of embryo developmental failures in mammals. The occurrence of these types of abnormalities is higher in in vitro-produced (IVP) embryos. The aim of the present study was to investigate the effect of oocyte morphology and maturation conditions on the rate of chromosomal abnormalities in bovine preimplantational embryos. To this end, 790 early cattle embryos derived from oocytes with different morphologies and matured under different conditions, including maturation period (24 v. 36h) and maturation media (five different serum supplements in TCM-199), were evaluated cytogenetically in three sequential experiments. The rates of normal diploidy and abnormal haploidy, polyploidy and aneuploidy were determined in each embryo. Throughout all the experiments, the rate of chromosomal abnormalities was significantly (P<0.05) affected by oocyte morphology and maturation conditions (maturation time and culture medium). Lower morphological quality was associated with a high rate of chromosome abnormalities (P<0.05). Moreover, polyploidy was associated with increased maturation time (P<0.01), whereas the maturation medium significantly (P<0.05) affected the rates of haploidy and polyploidy. In general, supplementing the maturation medium with oestrous cow serum or fetal calf serum resulted in higher rates of chromosomal aberrations (P<0.05) compared with the other serum supplements tested (bovine steer serum, anoestroues cow serum, bovine amniotic fluid and bovine serum albumin). On the basis of the results of the present study, we conclude that the morphological quality of oocytes and the maturation conditions affect the rate of chromosomal abnormalities in IVP bovine embryos.

  5. ‘Vanishing embryo syndrome’ in IVF/ICSI

    DEFF Research Database (Denmark)

    Hvidtjørn, Dorte; Grove, Jakob; Schendel, Diana;

    2005-01-01

    BACKGROUND: In a Danish population-based cohort study assessing the risk of cerebral palsy in children bornafter IVF, we made some interesting observations regarding ‘vanishing co-embryos’. METHODS andRESULTS: All live-born children born in Denmark from 1 January 1995 to 31 December 2000 were...... included inthis analysis. The children conceived by IVF/ICSI (9444) were identified through the IVF Register, the childrenconceived without IVF/ICSI (395 025) were identified through The Danish Medical Birth Register. Main outcomemeasure was the incidence of cerebral palsy. Within the IVF/ICSI children we...... found indications of an increasedrisk of cerebral palsy in those children resulting from pregnancies, where the number of embryos transferred washigher than the number of children born. CONCLUSIONS: The association between vanishing embryo syndromeand incidence of cerebral palsy following IVF requires...

  6. IVF and embryo transfer: historical origin and development.

    Science.gov (United States)

    Biggers, John D

    2012-08-01

    IVF and embryo transfer for the treatment of human infertility has now resulted in the birth of over 4 million babies. The technique did not arise as a quantum event but was built on the efforts of many earlier workers in the fields of reproductive endocrinology and development. One should remember the famous saying of Isaac Newton: 'If I have seen further than most, it is because I have stood on the shoulder's of giants'. Ethical and moral issues have always arisen when investigators study early mammalian development, particularly human development. This paper documents these earlier studies and also draws attention to the ethical and moral arguments that inevitably arose. PMID:22695311

  7. Reducing twin pregnancy rates after IVF--elective single embryo transfer (eSET).

    LENUS (Irish Health Repository)

    Milne, P

    2010-01-01

    Multiple pregnancy is a major complication of IVF and is associated with increased maternal, fetal and neonatal morbidity. Elective single embryo transfer (eSET) during IVF, rather than the more standard transfer of two embryos (double embryo transfer or DET), has been shown to significantly reduce the multiple pregnancy rate associated with IVF, while maintaining acceptable pregnancy rates. Couples undergoing IVF in 2008 who met good prognostic criteria had eSET performed. Pregnancy and twinning rates were compared with those for similar couples in 2007 who had DET. Couples unsuccessful with a fresh cycle of treatment had subsequent frozen embryo transfer cycles with DET. The cumulative pregnancy rate was similar for each group. However there were no multiple pregnancies in the eSET group, compared to 4 twins of 5 pregnancies in the DET group. 96% of eligible couples agreed to eSET. ESET is successful in and acceptable to good prognosis Irish couples undergoing IVF.

  8. Accurate and noninvasive embryos screening during in vitro fertilization (IVF) assisted by Raman analysis of embryos culture medium Accurate and noninvasive embryos screening during IVF

    Science.gov (United States)

    Shen, A. G.; Peng, J.; Zhao, Q. H.; Su, L.; Wang, X. H.; Hu, J. M.; Yang, J.

    2012-04-01

    In combination with morphological evaluation tests, we employ Raman spectroscopy to select higher potential reproductive embryos during in vitro fertilization (IVF) based on chemical composition of embryos culture medium. In this study, 57 Raman spectra are acquired from both higher and lower quality embryos culture medium (ECM) from 10 patients which have been preliminarily confirmed by clinical assay. Data are fit by using a linear combination model of least squares method in which 12 basis spectra represent the chemical features of ECM. The final fitting coefficients provide insight into the chemical compositions of culture medium samples and are subsequently used as criterion to evaluate the quality of embryos. The relative fitting coefficients ratios of sodium pyruvate/albumin and phenylalanine/albumin seem act as key roles in the embryo screening, attaining 85.7% accuracy in comparison with clinical pregnancy. The good results demonstrate that Raman spectroscopy therefore is an important candidate for an accurate and noninvasive screening of higher quality embryos, which potentially decrease the time-consuming clinical trials during IVF.

  9. Bovine in vitro embryo production : An overview

    Directory of Open Access Journals (Sweden)

    V. S. Suthar

    Full Text Available Dairy industry perfected the application of the first reproductive biotechnology, i.e. artificial insemination (AI - a great success story and also remains the user of embryo transfer technology (ETT. In addition, recently the researchers taking interest to embraced the field of Transvaginal OocyteRecovery (TVOR and in vitro production (IVEP of embryos. IVF provides the starting point for the generation of reproductive material for a number of advanced reproduction techniques including sperm microinjection and nuclear transfer (cloning. In several countries commercial IVF facilities are already being employed by cattle ET operators. Various research groups have reported on modification of TVOR technique to give greater efficiency. Much research is still needed in domestic animal (Especially Indian species on mechanisms controlling embryo development and on development of totally in vitro system for embryo culture. [Vet World 2009; 2(12.000: 478-479`

  10. Accuracy of a combined score of zygote and embryo morphology for selecting the best embryos for IVF

    Institute of Scientific and Technical Information of China (English)

    Yu-li QIAN; Ying-hui YE; Chen-ming XU; Fan JIN; He-feng HUANG

    2008-01-01

    Objective:To evaluate the accuracy of a scoring system combining zygote and embryo morphology in predicting the outcome of in vitro fertilization(IVF)treatment.Methods:In a study group,117 consecutive IVF or intracytoplasmic sperm injection(ICSI) cycles with embryo transfer were carried out and 312 embryos were scored Using a combmed scoring system(CSS)of zygote and embryo morphology before transplantation.In a control group,a total of 420 IVF or ICSI cycles were carried out and 1176 embryos were scored using a cumulative embryo score(CES).The effects of the combined scoring system on the embryo implantation rate and pregnancy rate per cycle were analyzed.Results:Using the combined scoring system,the embryo implantation rate(27.6%)and the clinical pregnancy rate(48.7%)were significantly higher than those in the control group(20.8%and 38.6%,respectively).Also,the implantation rate of embryos scoring≥70 (38.5%:82 sacs/213 embryos)was significantly higher (P<0.001)than that of embryos scoring<70(4%:4 sacs/99 embryos).The pregnancy rate of patients with embryos scoring≥70 using the combined scoring system(66.7%)Was significantly higher(P<0.001)than that of patients with embryos scoring≥20 using the cumulative embryo score(59.0%).Conclusion:The results suggest that selecting embryos with a high Score(≥70)using the combined scoring system could inerease the implantation rate and pregnancy rate,and that using a scoring system combining assessments of human zygotes and pre-implantation embryos might predict IVF outcomes more accurately than using a cumulafive embryo score.

  11. Genomic DNA methylation patterns in bovine preimplantation embryos derived from in vitro fertilization

    Institute of Scientific and Technical Information of China (English)

    HOU Jian; LIU Lei; LEI TingHua; CUI XiuHong; AN XiaoRong; CHEN YongFu

    2007-01-01

    By using the approach of immunofluorescence staining with an antibody against 5-methylcytosine (5MeC), the present study detected the DNA methylation patterns of bovine zygotes and preimplantation embryos derived from oocyte in vitro maturation (IVM), in vitro fertilization (IVF) and embryo in vitro culture (IVC). The results showed that: a) paternal-specific demethylation occurred in 61.5% of the examined zygotes, while 34.6% of them showed no demethylation; b) decreased methylation level was observed after the 8-cell stage and persisted through the morula stage, however methylation levels were different between blastomeres within the same embryos; c) at the blastocyst stage, the methylation level was very low in inner cell mass, but high in trophectoderm cells. The present study suggests, at least partly, that IVM/IVF/IVC may have effects on DNA methylation reprogramming of bovine zygotes and early embryos.

  12. Genomic DNA methylation patterns in bovine preim-plantation embryos derived from in vitro fertilization

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    By using the approach of immunofluorescence staining with an antibody against 5-methylcytosine (5MeC), the present study detected the DNA methylation patterns of bovine zygotes and preimplanta-tion embryos derived from oocyte in vitro maturation (IVM), in vitro fertilization (IVF) and embryo in vitro culture (IVC). The results showed that: a) paternal-specific demethylation occurred in 61.5% of the examined zygotes, while 34.6% of them showed no demethylation; b) decreased methylation level was observed after the 8-cell stage and persisted through the morula stage, however methylation levels were different between blastomeres within the same embryos; c) at the blastocyst stage, the methyla-tion level was very low in inner cell mass, but high in trophectoderm cells. The present study suggests, at least partly, that IVM/IVF/IVC may have effects on DNA methylation reprogramming of bovine zygotes and early embryos.

  13. Cleavage speed and implantation potential of early-cleavage embryos in IVF or ICSI cycles

    OpenAIRE

    Lee, Meng-Ju; Lee, Robert Kuo-Kuang; Lin, Ming-Huei; Hwu, Yuh-Ming

    2012-01-01

    We examined whether there is a correlation among early embryo cleavage, speed of cleavage, and implantation potential for in-vitro fertilization (IVF) treatment and intracytoplasmic sperm injection (ICSI). This retrospective study examined 112 cycles of IVF and 82 cycles of ICSI in patients less than 40 years of age. Early cleavage was defined as embryonic mitosis occurring 25–27 h after insemination. These day-3 embryos were then grouped according to cleavage speed (rapid, normal, and slow) ...

  14. Oocyte-secreted factors in oocyte maturation media enhance subsequent development of bovine cloned embryos.

    Science.gov (United States)

    Su, Jianmin; Wang, Yongsheng; Zhang, Lei; Wang, Bo; Liu, Jun; Luo, Yan; Guo, Zekun; Quan, Fusheng; Zhang, Yong

    2014-04-01

    Successful in vitro maturation (IVM) and oocyte quality both affect the subsequent development of cloned embryos derived from somatic-cell nuclear transfer (SCNT). Developmental competence is usually lower in oocytes matured in vitro compared with those that matured in vivo, possibly due to insufficient levels of oocyte-secreted factors (OSFs) and disrupted oocyte-cumulus communication. This study investigated the effects of OSFs secreted by denuded oocytes (DOs) during IVM on the subsequent developmental competence of cloned bovine embryos. Cumulus-oocyte complexes (COCs) from antral follicles of slaughtered-cow ovaries collected from an abattoir were divided into four groups: COCs co-cultured with and without DOs in maturation media used for SCNT, as well as COCs co-cultured with and without DOs in maturation media used for in vitro fertilization (IVF). Based on the developmental competence and embryo quality of bovine embryos generated from these four groups, we found that co-culturing the COCs with DOs enhanced the in vitro development of IVF and cloned bovine embryos, and potentially generated more high-quality cloned blastocysts that possessed locus-specific histone modifications at levels similar to in vitro-fertilized embryos. These results strongly suggest that co-culturing COCs with DOs enhances subsequent developmental competence of cloned bovine embryo. PMID:24420374

  15. LIF and TNF alpha concentrations in embryo culture media are predictive for embryo implantation in IVF

    Institute of Scientific and Technical Information of China (English)

    Ursula Zollner; Sonja Bischofs; Irena Lalic; Klaus-Peter Zollner

    2012-01-01

    Objective:There is strong evidence that the cytokines leucemia inhibitory factor (LIF) and tumor necrosis factor (TNF) alpha are related to embryo development and implantation. The aim of this study was to determine the levels of LIF and TNF alpha in embryo culture media and to assess its relationship to the outcome of in-vitro fertilization and embryo transfer. Methods:A total of 99 patients were included in this prospective trial and underwent either IVF or ICSI procedure. A total of 865 oocytes were collected. Embryos were cultured in sequential media until day 5. A standardized morphology evaluation of all embryos, including a detailed pronuclear scoring, was performed daily during this period followed by the replacement of one or two selected embryos. Collected embryo culture fluids of days 3 and 5 were analysed for LIF and TNF alpha on days 3 and 5. Results:Mean TNF alpha concentration in culture media on day 3 was 0.54 and 0.37 pg/mL on day 5 and was significantly lower in women conceiving than in not conceiving (0.43 pg/mL versus 0.59 pg/mL on day 3). Mean LIF concentration on day 3 was 31.5 pg/mL and 35.5 pg/mL on day 5 and was significantly higher in women conceiving (56.2 pg/mL versus 22.2 pg/mL on day 3). Conclusions:The results indicate that LIF could have a function in early embryogenesis and as a factor required for embryo implantation. High TNF alpha concentrations seem to be predictive of implantation failure.

  16. IVF outcomes in average- and poor-prognosis infertile women according to the number of embryos transferred.

    Science.gov (United States)

    Vega, Mario G; Gleicher, Norbert; Darmon, Sarah K; Weghofer, Andrea; Wu, Yan-Guang; Wang, Qi; Zhang, Lin; Albertini, David F; Barad, David H; Kushnir, Vitaly A

    2016-09-01

    Outcome measures of IVF success, which account for effectiveness of IVF and perinatal outcome risks, have recently been described. The association between number of embryos transferred in average and poor-prognosis IVF patients, and the chances of having good or poor IVF and perinatal outcomes, was investigated. Good IVF and perinatal outcome was defined as the birth of a live, term, normal-weight infant (≥2500 g). Poor IVF and perinatal outcome was defined as no live birth or birth of a very low weight neonate (<1500 g) or severe prematurity (birth at <32 weeks gestation). Each neonate was analysed as a separate outcome. A total of 713 IVF cycles in 504 average and poor-prognosis patients from January 2010 to December 2013 were identified. The odds of having good IVF and perinatal outcomes increased by 28% for each additional embryo transferred. The odds of poor IVF and perinatal outcome decreased by 32% with an additional embryo transferred. The likelihood of live birth with good perinatal outcome in average- and poor-prognosis patients after IVF increases with additional embryos being transferred. These data add to recently reported evidence in favour of multiple embryo transfer in older women and those with average or poor IVF prognosis.

  17. In vitro production of bovine embryos

    DEFF Research Database (Denmark)

    Stroebech, L.; Mazzoni, Gianluca; Pedersen, Hanne Skovsgaard;

    2015-01-01

    In vitro production (IVP) of bovine embryos has become a widespread technology implemented in cattle breeding and production. The implementation of genomic selection and systems biology adds great dimensions to the impact of bovine IVP. The physical procedures included in the IVP process can still...

  18. PreImplantation Factor (PIF correlates with early mammalian embryo development-bovine and murine models

    Directory of Open Access Journals (Sweden)

    Coulam Carolyn B

    2011-05-01

    Full Text Available Abstract Background PreImplantation Factor (PIF, a novel peptide secreted by viable embryos is essential for pregnancy: PIF modulates local immunity, promotes decidual pro-adhesion molecules and enhances trophoblast invasion. To determine the role of PIF in post-fertilization embryo development, we measured the peptide's concentration in the culture medium and tested endogenous PIF's potential trophic effects and direct interaction with the embryo. Methods Determine PIF levels in culture medium of multiple mouse and single bovine embryos cultured up to the blastocyst stage using PIF-ELISA. Examine the inhibitory effects of anti-PIF-monoclonal antibody (mAb added to medium on cultured mouse embryos development. Test FITC-PIF uptake by cultured bovine blastocysts using fluorescent microscopy. Results PIF levels in mouse embryo culture medium significantly increased from the morula to the blastocyst stage (ANOVA, P = 0.01. In contrast, atretic embryos medium was similar to the medium only control. Detectable - though low - PIF levels were secreted already by 2-cell stage mouse embryos. In single bovine IVF-derived embryos, PIF levels in medium at day 3 of culture were higher than non-cleaving embryos (control (P = 0.01 and at day 7 were higher than day 3 (P = 0.03. In non-cleaving embryos culture medium was similar to medium alone (control. Anti-PIF-mAb added to mouse embryo cultures lowered blastocyst formation rate 3-fold in a dose-dependent manner (2-way contingency table, multiple groups, X2; P = 0.01 as compared with non-specific mouse mAb, and medium alone, control. FITC-PIF was taken-up by cultured bovine blastocysts, but not by scrambled FITC-PIF (control. Conclusions PIF is an early embryo viability marker that has a direct supportive role on embryo development in culture. PIF-ELISA use to assess IVF embryo quality prior to transfer is warranted. Overall, our data supports PIF's endogenous self sustaining role in embryo development and the

  19. Single embryo transfer and IVF/ICSI outcome: a balanced appraisal.

    Science.gov (United States)

    Gerris, Jan M R

    2005-01-01

    This review considers the value of single embryo transfer (SET) to prevent multiple pregnancies (MP) after IVF/ICSI. The incidence of MP (twins and higher order pregnancies) after IVF/ICSI is much higher (approximately 30%) than after natural conception (approximately 1%). Approximately half of all the neonates are multiples. The obstetric, neonatal and long-term consequences for the health of these children are enormous and costs incurred extremely high. Judicious SET is the only method to decrease this epidemic of iatrogenic multiple gestations. Clinical trials have shown that programmes with >50% of SET maintain high overall ongoing pregnancy rates ( approximately 30% per started cycle) while reducing the MP rate to <10%. Experience with SET remains largely European although the need to reduce MP is accepted worldwide. An important issue is how to select patients suitable for SET and embryos with a high putative implantation potential. The typical patient suitable for SET is young (aged <36 years) and in her first or second IVF/ICSI trial. Embryo selection is performed using one or a combination of embryo characteristics. Available evidence suggests that, for the overall population, day 3 and day 5 selection yield similar results but better than zygote selection results. Prospective studies correlating embryo characteristics with documented implantation potential, utilizing databases of individual embryos, are needed. The application of SET should be supported by other measures: reimbursement of IVF/ICSI (earned back by reducing costs), optimized cryopreservation to augment cumulative pregnancy rates per oocyte harvest and a standardized format for reporting results. To make SET the standard of care in the appropriate target group, there is a need for more clinical studies, for intensive counselling of patients, and for an increased sense of responsibility in patients, health care providers and health insurers.

  20. Effects of steroidal glycoalkaloids from potatoes (Solanum tuberosum) on in vitro bovine embryo development.

    Science.gov (United States)

    Wang, S; Panter, K E; Gaffield, W; Evans, R C; Bunch, T D

    2005-02-01

    alpha-Solanine and alpha-chaconine are two naturally occurring steroidal glycoalkaloids in potatoes (Solanum tuberosum), and solanidine-N-oxide is a corresponding steroidal aglycone. The objective of this research was to screen potential cyto-toxicity of these potato glycoalkaloids using bovine oocyte maturation, in vitro fertilization techniques and subsequent embryonic development as the in vitro model. A randomized complete block design with four in vitro oocyte maturation (IVM) treatments (Experiment 1) and four in vitro embryo culture (IVC) treatments (Experiment 2) was used. In Experiment 1, bovine oocytes (n=2506) were matured in vitro in medium supplemented with 6 microM of alpha-solanine, alpha-chaconine, solanidine-N-oxide or IVM medium only. The in vitro matured oocytes were then subject to routine IVF and IVC procedures. Results indicated that exposure of bovine oocytes to the steroidal glycoalkaloids during in vitro maturation inhibited subsequent pre-implantation embryo development. Potency of the embryo-toxicity varied between these steroidal glycoalkaloids. In Experiment 2, IVM/IVF derived bovine embryos (n=2370) were cultured in vitro in medium supplemented with 6 microM of alpha-solanine, alpha-chaconine, solanidine-N-oxide or IVC medium only. The results showed that the pre-implantation embryo development is inhibited by exposure to these glycoalkaloids. This effect is significant during the later pre-implantation embryo development period as indicated by fewer numbers of expanded and hatched blastocysts produced in the media containing these alkaloids. Therefore, we conclude that in vitro exposure of oocytes and fertilized ova to the steroidal glycoalkaloids from potatoes inhibits pre-implantation embryo development. Furthermore, we suggest that ingestion of Solanum species containing toxic amounts of glycoalkaloids may have negative effects on pre-implantation embryonic survival.

  1. Presence of bile acids in human follicular fluid and their relation with embryo development in modified natural cycle IVF

    NARCIS (Netherlands)

    Nagy, R. A.; van Montfoort, A. P. A.; Dikkers, A.; van Echten-Arends, J.; Homminga, I.; Land, J. A.; Hoek, A.; Tietge, U. J. F.

    2015-01-01

    STUDY QUESTION: Are bile acids (BA) and their respective subspecies present in human follicular fluid (FF) and do they relate to embryo quality in modified natural cycle IVF (MNC-IVF)? SUMMARY ANSWER: BAconcentrations are 2-fold higher in follicular fluid than in serum and ursodeoxycholic acid (UDCA

  2. Accurate and noninvasive embryos screening during in vitro fertilization (IVF) assisted by Raman analysis of embryos culture medium

    International Nuclear Information System (INIS)

    In combination with morphological evaluation tests, we employ Raman spectroscopy to select higher potential reproductive embryos during in vitro fertilization (IVF) based on chemical composition of embryos culture medium. In this study, 57 Raman spectra are acquired from both higher and lower quality embryos culture medium (ECM) from 10 patients which have been preliminarily confirmed by clinical assay. Data are fit by using a linear combination model of least squares method in which 12 basis spectra represent the chemical features of ECM. The final fitting coefficients provide insight into the chemical compositions of culture medium samples and are subsequently used as criterion to evaluate the quality of embryos. The relative fitting coefficients ratios of sodium pyruvate/albumin and phenylalanine/albumin seem act as key roles in the embryo screening, attaining 85.7% accuracy in comparison with clinical pregnancy. The good results demonstrate that Raman spectroscopy therefore is an important candidate for an accurate and noninvasive screening of higher quality embryos, which potentially decrease the time-consuming clinical trials during IVF

  3. Nucleolar ultrastructure in bovine nuclear transfer embryos

    DEFF Research Database (Denmark)

    Kaňka, Jiří; Smith, Steven Dale; Soloy, Eva;

    1999-01-01

    in nuclear morphology as a transformation of the nucleolus precursor body into a functional rRNA synthesising nucleolus with a characteristic ultrastructure. We examined nucleolar ultrastructure in bovine in vitro produced (control) embryos and in nuclear transfer embryos reconstructed from a MII phase...... at 1 hr after fusion and, by 3 hr after fusion, it was restored again. At this time, the reticulated fibrillo-granular nucleolus had an almost round shape. The nucleolar precursor body seen in the two-cell stage nuclear transfer embryos consisted of intermingled filamentous components and secondary...... time intervals after fusion. In the two-cell stage nuclear transfer embryo, the originally reticulated nucleolus of the donor blastomere had changed into a typical nucleolar precursor body consisting of a homogeneous fibrillar structure. A primary vacuole appeared in the four-cell stage nuclear...

  4. Transcriptional reprogramming of gene expression in bovine somatic cell chromatin transfer embryos

    Directory of Open Access Journals (Sweden)

    Page Grier P

    2009-04-01

    Full Text Available Abstract Background Successful reprogramming of a somatic genome to produce a healthy clone by somatic cells nuclear transfer (SCNT is a rare event and the mechanisms involved in this process are poorly defined. When serial or successive rounds of cloning are performed, blastocyst and full term development rates decline even further with the increasing rounds of cloning. Identifying the "cumulative errors" could reveal the epigenetic reprogramming blocks in animal cloning. Results Bovine clones from up to four generations of successive cloning were produced by chromatin transfer (CT. Using Affymetrix bovine microarrays we determined that the transcriptomes of blastocysts derived from the first and the fourth rounds of cloning (CT1 and CT4 respectively have undergone an extensive reprogramming and were more similar to blastocysts derived from in vitro fertilization (IVF than to the donor cells used for the first and the fourth rounds of chromatin transfer (DC1 and DC4 respectively. However a set of transcripts in the cloned embryos showed a misregulated pattern when compared to IVF embryos. Among the genes consistently upregulated in both CT groups compared to the IVF embryos were genes involved in regulation of cytoskeleton and cell shape. Among the genes consistently upregulated in IVF embryos compared to both CT groups were genes involved in chromatin remodelling and stress coping. Conclusion The present study provides a data set that could contribute in our understanding of epigenetic errors in somatic cell chromatin transfer. Identifying "cumulative errors" after serial cloning could reveal some of the epigenetic reprogramming blocks shedding light on the reprogramming process, important for both basic and applied research.

  5. Demi-embryo production from hatching of zona-drilled bovine and rabbit blastocysts.

    Science.gov (United States)

    Skrzyszowska, M; Smorag, Z; Katska, L

    1997-09-01

    It is known that the pregnancy rate resulting after transfer of bisected embryos is lower than after transfer of whole embryos. The main reason is the reduced cell number in the demi-embryo which is less than 1 2 of that in the intact embryo, since a number of blastomeres is damaged as a result of the procedure used in conventional embryo splitting. The aim of our experiment was to develop a non-invasive procedure which would limit cell losses during microsurgery. The experiment was carried out on bovine IVM-IVF embryos at middle, late and expanded blastocyst stage and rabbit embryos at late blastocyst stage cultured in vitro from in vivo produced zygotes. The zona pellucida of these embryos was drilled on the line between the inner cell mass and the trophoblast using a glass microneedle (embryo configuration, connected by a very thin cell bridge (figure eight in shape). To separate the parts of the embryo, the cell bridge was cut using a glass microneedle. During the separation only a few cells were damaged. As a result of the procedure 4 20 (20.0%), 48 144 (33.3%) and 3 40 (7.5%) middle, late and expanded blastocysts hatched according to the pattern described. The developed procedure could be considered as a non-invasive alternative to conventional embryo splitting.

  6. Nucleologenesis and embryonic genome activation are defective in interspecies cloned embryos between bovine ooplasm and rhesus monkey somatic cells

    Directory of Open Access Journals (Sweden)

    Han Yong-Mahn

    2009-07-01

    Full Text Available Abstract Background Interspecies somatic cell nuclear transfer (iSCNT has been proposed as a tool to address basic developmental questions and to improve the feasibility of cell therapy. However, the low efficiency of iSCNT embryonic development is a crucial problem when compared to in vitro fertilization (IVF and intraspecies SCNT. Thus, we examined the effect of donor cell species on the early development of SCNT embryos after reconstruction with bovine ooplasm. Results No apparent difference in cleavage rate was found among IVF, monkey-bovine (MB-iSCNT, and bovine-bovine (BB-SCNT embryos. However, MB-iSCNT embryos failed to develop beyond the 8- or 16-cell stages and lacked expression of the genes involved in embryonic genome activation (EGA at the 8-cell stage. From ultrastructural observations made during the peri-EGA period using transmission electron microscopy (TEM, we found that the nucleoli of MB-iSCNT embryos were morphologically abnormal or arrested at the primary stage of nucleologenesis. Consistent with the TEM analysis, nucleolar component proteins, such as upstream binding transcription factor, fibrillarin, nucleolin, and nucleophosmin, showed decreased expression and were structurally disorganized in MB-iSCNT embryos compared to IVF and BB-SCNT embryos, as revealed by real-time PCR and immunofluorescence confocal laser scanning microscopy, respectively. Conclusion The down-regulation of housekeeping and imprinting genes, abnormal nucleolar morphology, and aberrant patterns of nucleolar proteins during EGA resulted in developmental failure in MB-iSCNT embryos. These results provide insight into the unresolved problems of early embryonic development in iSCNT embryos.

  7. Splitting and biopsy for bovine embryo sexing under field conditions.

    Science.gov (United States)

    Lopes, R F; Forell, F; Oliveira, A T; Rodrigues, J L

    2001-12-01

    Improvements on embryo micromanipulation techniques led to the use of embryo bisection technology in commercial embryo transfer programs, and made possible the direct genetic analysis of preimplantation bovine embryos by biopsy. For example, aspiration and microsection, allow bovine embryos sexing by detection of male-specific Y-chromosome in a sample of embryonic cells. We report on the application of the methodologies of splitting and biopsy of bovine embryos in field conditions, and on the results of embryo sex determination by the polymerase chain reaction (PCR). Pregnancy rates achieved with fresh bisected or biopsied embryos (50 to 60%) were similar to the fresh intact embryos (55 to 61%). The PCR protocol used for embryo sexing showed 92% to 94% of efficiency and 90 to 100% of accuracy. These results demonstrate these procedures are suitable for use in field conditions.

  8. Effect of the P1 Medium and the ECM Medium on Embryo Quality in IVF

    Institute of Scientific and Technical Information of China (English)

    Qian CHEN; Ai-jun ZHANG; Yun FENG; Xiao-wei LU; Dong-mei JI; Zhi-peng XU

    2009-01-01

    Objective To investigate the effect of the glucose-free reimplantation stage one(P1) medium and the ECM medium on embryo development quality in IVF.Methods The patients with ≥4 zygotes of 2PN were studied.A total of 201 retrieval cycles were included in a prospective randomized study.Each patient was herself control Half of zygotes of 2PN were transferred into ECM medium(group A)and half into P1 medium(group B)for further culture.Embryo development was evaluated on the day of embryo transfer.The efficacy of ECM was compared with P1 as culture medium for the development of preimplantation embryos. Results No statistically significant differences were noted between the two groups regarding embryo-cleavage rate(97.13% vs 97.55%)and rate of normal-cleaving embryos(58.29% and 58.37%).The rate of top-quality embryos was statistically higher in group A than in group B(27.59% vs 19.75%,P<0.05).Embryo quality,as assessed by morphological parameters(the amount of detached anuclear fragments>30%),was better in group A than in group B(19.86% vs 21.75%),however,there was no statistically significance.Both the rate of good-quality embryos(47.95% vs 46.17%)and available embryos(63.22% vs 61.,9%)were higher in group A than in group B,but there was also no statistically significance.Conclusion The ECM medium may be associated with a better embryo quality compared with the P1 medium.

  9. Biopsy of human morula-stage embryos: outcome of 215 IVF/ICSI cycles with PGS.

    Directory of Open Access Journals (Sweden)

    Elena E Zakharova

    Full Text Available Preimplantation genetic diagnosis (PGD is commonly performed on biopsies from 6-8-cell-stage embryos or blastocyst trophectoderm obtained on day 3 or 5, respectively. Day 4 human embryos at the morula stage were successfully biopsied. Biopsy was performed on 709 morulae from 215 ICSI cycles with preimplantation genetic screening (PGS, and 3-7 cells were obtained from each embryo. The most common vital aneuploidies (chromosomes X/Y, 21 were screened by fluorescence in situ hybridization (FISH. No aneuploidy was observed in 72.7% of embryos, 91% of those developed to blastocysts. Embryos were transferred on days 5-6. Clinical pregnancy was obtained in 32.8% of cases, and 60 babies were born. Patients who underwent ICSI/PGS treatment were compared with those who underwent standard ICSI treatment by examining the percentage of blastocysts, pregnancy rate, gestational length, birth height and weight. No significant differences in these parameters were observed between the groups. Day 4 biopsy procedure does not adversely affect embryo development in vitro or in vivo. The increased number of cells obtained by biopsy of morulae might facilitate diagnostic screening. There is enough time after biopsy to obtain PGD results for embryo transfer on day 5-6 in the current IVF cycle.

  10. Evaluation of day one embryo quality and IVF outcome – a comparison of two scoring systems

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    Svobodova Magda

    2009-02-01

    Full Text Available Abstract Background The aim of our retrospective study was to compare the clinical usefulness of two non-invasive embryo scoring systems based either on a simplified pronuclear morphology of the zygote or on early cleavage rate, as well as their combination, for the selection of embryos with the best implantation potential in embryo transfer (ET. Methods Over a period of five years, the quality of 2708 embryos from 364 IVF cycles in women under the age of 39 years was assessed using these scoring systems in a university assisted reproduction centre. ET was always performed on day 3 of cultivation. The outcome of ETs of 702 embryos scored in the respective systems or their combination was retrospectively analyzed in terms of biochemical (bPR and clinical pregnancy rates (cPR and implantation rate (IR. Mann-Whitney U test and t-test for differences between relative values were used, p Results There was no difference in outcome parameters in 109 cycles where only Pattern "0" zygotes, according to our simplified pronuclear morphology classification, were transferred and 140 cycles where only "other" pattern zygotes were transferred, regardless of their cleavage rate. On the contrary, significantly greater cPR and IR (p = 0.003 and p = 0.006, respectively were achieved in 120 cycles where only early cleavage (EC embryos were transferred compared with 152 cycles where only non early cleavage (NEC embryos were transferred regardless of their pronuclear morphology. The best outcome in terms of cPR (56% and IR (43% was found in 50 cycles when Pattern "0" and EC embryos only were used for transfer. Conclusion The results indicate that early cleavage is a better independent marker of implantation potential than zygote morphology. The best outcome can be achieved if both embryo scoring systems are used jointly and the embryo is classified as EC and Pattern "0".

  11. Aneuploidy analysis of non-pronuclear embryos from IVF with use of array CGH: a case report.

    Science.gov (United States)

    Lixin, Deng; Zhifeng, Xiang; Cong, He; Jinzhou, Zhang; Hongbin, Xie

    2014-06-01

    By using array comparative genomic hybridization (array CGH), to analyze the aneuploidy of the single blastomeres from non-pronuclear embryos on cleavage-stage in IVF cycle. Four non-pronuclear embryos were got from an IVF cycle, and the each single cell was biopsied from the four cleavage-stage embryos on the third day after the insemination which was investigated by using array CGH. After the biopsy, all the embryos continued to cleave, and lately entered the morula stage on the fifth day, just one embryo 3 was developed to early blastocyst stage on the sixth day. The four blastomere 24 chromosomes showed one X monomer and three normal XY diploids; the autosome chromosomes of blastomeres were abnormally gained or lost at different chromosome from four embryos, such as Embryo 1 : 49,X (-1, -5, -11, -19, -20, -21, -Y, +3, +6, +7, +8, +10, +13, +14, +16, +17, +18); Embryo 2 : 44,XY (-12, -15); Embryo 3: 47,XY (-3, -8, -9, -21, +7, +17, +18, +19, +20); Embryo 4 : 54,XY (+4, +7, +10, +12, +13, +16, +17, +22). With the use of the array CGH, the aneuploidy analysis could review the abnormal chromosomes of single blastomere from the non-pronuclear embryos, which can harbor the risk of abnormal sex chromosome and autosome chromosomes.

  12. Noninvasive Metabolomic Profiling of Human Embryo Culture Media Using a Simple Spectroscopy Adjunct to Morphology for Embryo Assessment in in Vitro Fertilization (IVF

    Directory of Open Access Journals (Sweden)

    Jiming Hu

    2013-03-01

    Full Text Available Embryo quality is crucial to the outcome of in vitro fertilization (IVF; however, the ability to precisely distinguish the embryos with higher reproductive potential from others is poor. Morphologic evaluation used to play an important role in assessing embryo quality, but it is somewhat subjective. The culture medium is the immediate environment of the embryos in vitro, and a change of the substances in the culture medium is possibly related to the embryo quality. Thus, the present study aims to determine whether metabolomic profiling of the culture medium using Raman spectroscopy adjunct to morphology correlates with the reproductive potential of embryos in IVF and, thus, to look for a new method of assessing embryo quality. Fifty seven spent media samples were detected by Raman spectroscopy. Combined with embryo morphology scores, we found that embryos in culture media with less than 0.012 of sodium pyruvate and more than −0.00085 phenylalanine have a high reproductive potential, with up to 85.7% accuracy compared with clinical pregnancy. So, sodium pyruvate and phenylalanine in culture medium play an important role in the development of the embryo. Raman spectroscopy is an important tool that provides a new and accurate assessment of higher quality embryos.

  13. Meanings of the embryo in Japan: narratives of IVF experience and embryo ownership

    NARCIS (Netherlands)

    M. Kato; M. Sleeboom-Faulkner

    2011-01-01

    This article explores the sociocultural meanings of the embryo implied in the narratives of 58 women who have undergone in vitro fertilisation in Japan over a period from 2006 to 2008. We argue that a lack of sufficient analysis of the sociocultural meanings of the embryo result in a situation where

  14. Increased circulating cell-derived microparticle count is associated with recurrent implantation failure after IVF and embryo transfer.

    Science.gov (United States)

    Martínez-Zamora, M Angeles; Tàssies, Dolors; Reverter, Juan Carlos; Creus, Montserrat; Casals, Gemma; Cívico, Salvadora; Carmona, Francisco; Balasch, Juan

    2016-08-01

    Cell-derived microparticles (cMPs) are small membrane vesicles that are released from many different cell types in response to cellular activation or apoptosis. Elevated cMP counts have been found in almost all thrombotic diseases and pregnancy wastage, such as recurrent spontaneous abortion and in a number of conditions associated with inflammation, cellular activation and angiogenesis. cMP count was investigated in patients experiencing unexplained recurrent implantation failure (RIF). The study group was composed of 30 women diagnosed with RIF (RIF group). The first control group (IVF group) (n = 30) comprised patients undergoing a first successful IVF cycle. The second control group (FER group) included 30 healthy women who had at least one child born at term and no history of infertility or obstetric complications. cMP count was significantly higher in the RIF group compared with the IVF and FER groups (P < 0.05 and P < 0.01, respectively) (RIF group: 15.8 ± 6.2 nM phosphatidylserine equivalent [PS eq]; IVF group: 10.9 ± 5.3 nM PS eq; FER group: 9.6 ± 4.0 nM PS eq). No statistical difference was found in cMP count between the IVF and FER groups. Increased cMP count is, therefore, associated with RIF after IVF and embryo transfer.

  15. Elevated Progesterone Levels on the Day of Oocyte Maturation May Affect Top Quality Embryo IVF Cycles.

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    Bo Huang

    Full Text Available In contrast to the impact of elevated progesterone on endometrial receptivity, the data on whether increased progesterone levels affects the quality of embryos is still limited. This study retrospectively enrolled 4,236 fresh in vitro fertilization (IVF cycles and sought to determine whether increased progesterone is associated with adverse outcomes with regard to top quality embryos (TQE. The results showed that the TQE rate significantly correlated with progesterone levels on the day of human chorionic gonadotropin (hCG trigger (P = 0.009. Multivariate linear regression analysis of factors related to the TQE rate, in conventional IVF cycles, showed that the TQE rate was negatively associated with progesterone concentration on the day of hCG (OR was -1.658, 95% CI: -2.806 to -0.510, P = 0.005. When the serum progesterone level was within the interval 2.0-2.5 ng/ml, the TQE rate was significantly lower (P 2.5 ng/ml. Then, we choose a progesterone level at 1.5ng/ml, 2.0 ng/ml and 2.5 ng/ml as cut-off points to verify this result. We found that the TQE rate was significantly different (P 2.0 ng/ml. In conclusion, the results of this study clearly demonstrated a negative effect of elevated progesterone levels on the day of hCG trigger, on TQE rate, regardless of the basal FSH, the total gonadotropin, the age of the woman, or the time of ovarian stimulation. These data demonstrate that elevated progesterone levels (>2.0 ng/ml before oocyte maturation were consistently detrimental to the oocyte.

  16. Elevated Progesterone Levels on the Day of Oocyte Maturation May Affect Top Quality Embryo IVF Cycles

    Science.gov (United States)

    Huang, Bo; Ren, Xinling; Wu, Li; Zhu, Lixia; Xu, Bei; Li, Yufeng; Ai, Jihui; Jin, Lei

    2016-01-01

    In contrast to the impact of elevated progesterone on endometrial receptivity, the data on whether increased progesterone levels affects the quality of embryos is still limited. This study retrospectively enrolled 4,236 fresh in vitro fertilization (IVF) cycles and sought to determine whether increased progesterone is associated with adverse outcomes with regard to top quality embryos (TQE). The results showed that the TQE rate significantly correlated with progesterone levels on the day of human chorionic gonadotropin (hCG) trigger (P = 0.009). Multivariate linear regression analysis of factors related to the TQE rate, in conventional IVF cycles, showed that the TQE rate was negatively associated with progesterone concentration on the day of hCG (OR was -1.658, 95% CI: -2.806 to -0.510, P = 0.005). When the serum progesterone level was within the interval 2.0–2.5 ng/ml, the TQE rate was significantly lower (P 2.5 ng/ml. Then, we choose a progesterone level at 1.5ng/ml, 2.0 ng/ml and 2.5 ng/ml as cut-off points to verify this result. We found that the TQE rate was significantly different (P 2.0 ng/ml. In conclusion, the results of this study clearly demonstrated a negative effect of elevated progesterone levels on the day of hCG trigger, on TQE rate, regardless of the basal FSH, the total gonadotropin, the age of the woman, or the time of ovarian stimulation. These data demonstrate that elevated progesterone levels (>2.0 ng/ml) before oocyte maturation were consistently detrimental to the oocyte. PMID:26745711

  17. The relationship of the number of embryo transferred and IVF-ET outcome%胚胎移植的数量和IVF-ET结局的关系

    Institute of Scientific and Technical Information of China (English)

    杨晓丽; 张烁

    2009-01-01

    目的 探讨体外受精-胚胎移植(in vitro fertilization and embryo transfer, IVF-ET)过程中胚胎移植数从3个降至2个对IVF-ET结局的影响.方法 对161例35岁以下的不孕患者共161个IVF-ET周期的资料进行回顾性分析.将患者分为两组,其中移植2个胚胎的为第1组,移植3个胚胎的为第2组,从而比较不同胚胎移植数对临床妊娠率和多胎妊娠发生率的影响.结果 两组的临床妊娠率差异无统计学意义(P>0.05),但移植3个胚胎组的多胎妊娠率显著高于移植2个胚胎组(P<0.05).结论 35岁以下接受IVF的妇女,移植2个优质胚胎不影响妊娠的成功,同时可以降低多胎发生率.

  18. Evaluation of Bovine Embryo Biopsy Techniques according to Their Ability to Preserve Embryo Viability

    Directory of Open Access Journals (Sweden)

    M. Cenariu

    2012-01-01

    Full Text Available The purpose of this research was to evaluate three embryo biopsy techniques used for preimplantation genetic diagnosis (PGD in cattle and to recommend the least invasive one for current use, especially when PGD is followed by embryo cryopreservation. Three hundred bovine embryos were biopsied by either one of the needle, aspiration or microblade method, and then checked for viability by freezing/thawing and transplantation to recipient cows. The number of pregnancies obtained after the transfer of biopsied frozen/thawed embryos was assessed 30 days later using ultrasounds. The results were significantly different between the three biopsy methods: the pregnancy rate was of 57% in cows that received embryos biopsied by needle, 43% in cows that received embryos biopsied by aspiration, and 31% in cows that received embryos biopsied by microblade. Choosing an adequate biopsy method is therefore of great importance in embryos that will undergo subsequent cryopreservation, as it significantly influences their viability after thawing.

  19. Study of the morphology of ovary and cytology of oocyte as basis for establishing methods IVM, IVF and embryo transfer

    OpenAIRE

    Smiljaković T.; Poleksić V.; Petrović M.M.; Pejcić S.; Trenkovski S.; Stojanović Lj.; Alm H.

    2007-01-01

    In three types of domestic animals: cattle, pigs and sheep, morphology of ovary was studied. Results such as differences in shape, size within and between species, number of follicles in maturation, changes in sex cycle, connection of follicles in maturation are presented in figures. By method of aspiration, oocytes were isolated from ovaries and their cytology analyzed as the first step in methods of in vitro maturation (IVM), followed by in vitro fertilization (IVF) and embryo transfer. The...

  20. Coagulansin-A has beneficial effects on the development of bovine embryos in vitro via HSP70 induction

    Science.gov (United States)

    Khan, Imran; Lee, Kyeong-Lim; Fakruzzaman, Md.; Song, Seok-Hwan; Ihsan-ul-Haq; Mirza, Bushra; Yan, Chang Guo; Kong, Il-Keun

    2016-01-01

    Coagulansin-A (withanolide) is the steroidal lactone obtained from Withania coagulans which belong to Solanaceae family. The present study investigated the effects of coagulansin-A on bovine oocyte maturation and embryo development in vitro. All these oocytes were aspirated from the ovaries obtained from Korean Hanwoo cows at a local abattoir. To determine whether coagulansin-A has beneficial effects on bovine oocyte maturation in vitro, 355 oocytes per group (control and treated) in seven replicates were subjected with different concentrations (1, 2.5, 5, 7.5 and 10 μM) of coagulansin-A. The coagulansin-A was added in the in vitro maturation (IVM) media followed by in vitro fertilization (IVF) and then in vitro culture (IVC). Only treatment with 5 μM coagulansin-A remarkably (P<0.05) improved embryos development (Day 8 blastocyst) having 27.30 and 40.01% for control and coagulansin-A treated groups respectively. Treatment with 5 μM coagulansin-A significantly induced activation of heat shock protein 70 (HSP70) (P<0.05). Immunofluorescence analysis revealed that 5 μM coagulansin-A treatment also significantly inhibited oxidative stress and inflammation during bovine embryo development in vitro by decreasing 8-oxoguanosine (8-OxoG) (P<0.05) and nuclear factor-κB (NF-κB) (P<0.05). The expressions of HSP70 and NF-κB were also conformed through real-time PCR (RT-PCR). Additionally, the terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assay confirmed that coagulansin-A treatment significantly improved the embryo quality and reduced bovine embryo DNA damage (P<0.05). The present study provides new information regarding the mechanisms by which coagulansin-A promotes bovine embryo development in vitro. PMID:26831738

  1. Decisions for the IVF laboratory: comparative analysis of embryo culture incubators.

    Science.gov (United States)

    Swain, Jason E

    2014-05-01

    Incubators in the IVF laboratory play a pivotal role in providing a stable and appropriate culture environment required for optimizing embryo development and clinical outcomes. With technological advances, several types of incubators are now available and careful consideration is required for selection. Examination of variables, such as recovery/stabilization of temperature, gas atmosphere and humidity, as well as understanding various approaches utilized by each device to regulate these variables, is critical. Additionally, a comprehensive examination of clinical studies that compare various incubators may provide insight into their efficacy. Other factors, both technical and practical, must also be considered when selecting an incubator. Importantly, proper management, including patient volume and workflow, is paramount in optimizing function of any incubator, regardless of the technology incorporated. This review highlights incubator function and reviews key environmental variables controlled and the technology utilized in various units. Additionally, existing comparative studies focused on incubator recovery and clinical outcomes are critically analysed. Finally, strategies employed for incubator management, as well as future potential incubator improvements are discussed. While existing reports indicate that smaller benchtop/topload incubators provide faster recovery of environmental variables, there is no clear advantage of any particular incubator based on clinical outcomes.

  2. Long term costs and effects of reducing the number of twin pregnancies in IVF by single embryo transfer: the TwinSing study

    Directory of Open Access Journals (Sweden)

    van Goudoever Johannes B

    2010-10-01

    Full Text Available Abstract Background Pregnancies induced by in vitro fertilisation (IVF often result in twin gestations, which are associated with both maternal and perinatal complications. An effective way to reduce the number of IVF twin pregnancies is to decrease the number of embryos transferred from two to one. The interpretation of current studies is limited because they used live birth as outcome measure and because they applied limited time horizons. So far, research on long-term outcomes of IVF twins and singletons is scarce and inconclusive. The objective of this study is to investigate the short (1-year and long-term (5 and 18-year costs and health outcomes of IVF singleton and twin children and to consider these in estimating the cost-effectiveness of single embryo transfer compared with double embryo transfer, from a societal and a healthcare perspective. Methods/Design A multi-centre cohort study will be performed, in which IVF singletons and IVF twin children born between 2003 and 2005 of whom parents received IVF treatment in one of the five participating Dutch IVF centres, will be compared. Data collection will focus on children at risk of health problems and children in whom health problems actually occurred. First year of life data will be collected in approximately 1,278 children (619 singletons and 659 twin children. Data up to the fifth year of life will be collected in approximately 488 children (200 singletons and 288 twin children. Outcome measures are health status, health-related quality of life and costs. Data will be obtained from hospital information systems, a parent questionnaire and existing registries. Furthermore, a prognostic model will be developed that reflects the short and long-term costs and health outcomes of IVF singleton and twin children. This model will be linked to a Markov model of the short-term cost-effectiveness of single embryo transfer strategies versus double embryo transfer strategies to enable the

  3. Intra- and interobserver analysis in the morphological assessment of early stage embryos during an IVF procedure: a multicentre study

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    Devroe Johanna

    2011-09-01

    Full Text Available Abstract Background Quality control programs are necessary to maintain good clinical practice. Embryo grading has been described as one of the external quality assurance schemes. Although the evaluation of embryos is based on the assessment of morphological characteristics, considerable intra- and inter-observer variability has been described. In this multicentre study, the variability in the embryo evaluation has been evaluated using morphological characteristics on day 1, day 2 and day 3 of embryo development. Methods Five embryologists of four different IVF centers participated in this study. Multilevel images of embryos were presented on a website at different time points to evaluate intra-and inter-observer agreement in the assessment of embryo morphology. The embryos were evaluated on day 1, day 2 and day 3 of their development and each embryologist had to decide if the embryo had to be transferred, cryopreserved or discarded. Results Both intra-observer agreement and inter-observer agreement were good to excellent for the position of the pronuclei on day 1, the number of blastomeres on day 2 and day 3 and the clinical decision (transfer, cryopreservation, discard. For all other characteristics (size of pronuclei, presence of cytoplasomic halo, degree of fragmentation and size of blastomeres the intra- and inter-observer agreement was moderate to very poor. Conclusions Mono- or multicentre quality control on embryo scoring by morphological assessment can easily be performed through the design of a simple website. In the future the website design can be adapted to generate statistical feedback upon scoring and can even include a training module.

  4. IVF policy and global/local politics: the making of multiple-embryo transfer regulation in Taiwan.

    Science.gov (United States)

    Wu, Chia-Ling

    2012-08-01

    This paper analyzes the regulatory trajectory of multiple-embryo transfer in in-vitro fertilization (IVF) in Taiwan. Taking a latecomer to policy-making as the case, it argues the importance of conceptualizing the global/local dynamics in policy-making for assisted reproductive technology (ART). The conceptual framework is built upon recent literature on standardization, science policy, and global assemblage. I propose three interrelated features that reveal the "global in the local": (1) the power relationships among stakeholders, (2) the selected global form that involved actors drew upon, and (3) the re-contextualized assemblage made of local networks. Data included archives, interviews, and participant observation. In different historical periods the specific stakeholders selected different preferred global forms for Taiwan, such as Britain's code of ethics in the 1990s, the American guideline in the early 2000s, and the European trend in the mid-2000s. The global is heterogeneous. The failure to transfer the British regulation, the revision of the American guideline by adding one more embryo than it specified, and the gap between the cited European trend and the "no more than four" in Taiwan's 2007 Human Reproduction Law all show that the local network further transforms the selected global form, confining it to rhetoric only or tailoring it to local needs. Overall, Taiwanese practitioners successfully maintained their medical autonomy to build a 'flexible standardization'. Multiple pregnancy remains the most common health risk of IVF in Taiwan. PMID:22607747

  5. Research on Growth Behavior of Embryos for Bovine and Murine on Primary Murine Embryos Fibroblast Cell Feeder Layer

    Institute of Scientific and Technical Information of China (English)

    AN Li-long; XIAO Mei; FENG Xiu-Liang; DOU Zhong-ying; QIU Huai; YANG Qi; LEI An-min; YANG Chun-rong; GAO Zhi-min

    2002-01-01

    The difference in growth behavior between bovine embryos and murine embryos was studied on PMEF(primary murine embryos fibroblast)feeder layer. The results showed as follows: With embryos having attached, bovine embryonic trophoblast formed a transparent membranous structure covering on inner cell mass (ICM), however, murine embryonic trophoblast formed disc structure. Bovine embryos formed four kinds of ICM colonies with different morphology including the mass-like, the net-like, the stream-like and the mixture-like colonies. Compared with Murine ICM, the bovine ICM grew more fast. So, the bovine ICM was passaged at first after a culture of approximately 5 - 6 days in vitro, but murine ICM was passaged at first after an attachment of 3 - 4 days on PMEF feeder layer. The mixture colonies of bovine ICM differentiated very early, while the others differentiated very late. Most ICM-like mass of Bovine grew in a defined spot, but bovine ICMs like stream and ICMs like net proliferated fast and dispersed quickly. We found that the single blastomeres derived from late bovine morula and late murine morula formed sub-blastophere; moreover, the bovine ICM cell would differentiate rapidly if the trophoblast was removed.

  6. EFFECT OF FETAL SERUM AND FOLLICULAR LIQUOR SUPPLEMENTATION ON THE IN VITRO PRODUCTION OF BOVINE EMBRYOS

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    Clara Larocca

    2012-01-01

    Full Text Available To optimize In Vitro Fertilization (IVF results in bovines by lowering costs, comparing the efficiency of Fetal Calf Serum (FCS (high cost with respect to bovine Follicular Fluid (bFF (low cost and by quantifying embryo production. Cumulus-Oocyte-Complexes (COC obtained from a slaughter house, transported at 37°C in saline solution, were classified according to their quality in A, B, C and D. A and B oocytes were washed with modified Phosphate-Buffered Saline (m-PBS and three groups were randomly formed (GC; G1; G2 cultured in drops of the respective media (10 COC/drop of 100 μL, covered with mineral oil and placed in an incubator (38,5°C, 5% CO2 y 95% humidity. Maturation was done in Tissue Culture Medium (TCM-199 and antibiotics, for 22h and developed in CR1aa medium and antibiotics. The Control Group (CG was supplemented during both maturation and development stages with 5% FCS and 10% bFF; G1 with 5% FCS during maturation and development and G2 with 10% bFF during maturation and development. We obtained bFF from follicles larger than 15 mm, it was centrifuged (800G inactivating it. At fertilization Bracket and Oliphant (BO medium was used. Zygotes were evaluated every 48 h for 8 days since insemination, counting the Division Rate (DR and the Embryo Development (ED of compacted morulae. Χ2 Test was applied. For RD, G2 differs from G1 and CG (p0.05. During development no significant differences were found between groups (p>0.05. Results show that when using FCS, bFF or a combination of both, development results are similar. It is assumed that it is possible to substitute FCS with bFF."

  7. Relationship between Pronuclear Scoring and Embryo Quality and Implantation Potential in IVF-ET

    Institute of Scientific and Technical Information of China (English)

    Qun LIU; Guijin ZHU; Juan HU; Yulan WEI; Xinling REN; Hanwang ZHANG; Yufeng LI; Lei JIN; Jing YUE

    2008-01-01

    To assess the relationship between pronuclear scoring and day-3 embryo quality and pregnancy outcome and to determine the clinical value of pronuclear stage scoring system in human in vitro fertilization-embryo transfer (WF-ET) program, a pronuclear scoring system was used to score zygotes 16-20 h after insemination during conventional WF or intracytoplasmic sperm injec- tion (ICS1). The embryos were classified into groups Z1, Z2, Z3 and Z4. Comparisons were made of the rates of arrested embryos and excellent embryos on day 3. Comparisons of pregnancy outcome were made only in those patients in whom cohorts of similarly Z-scored embryos were transferred. The results showed that there were less arrested embryos and more excellent embryos on day 3 in groups Z1 and Z2 than those in group Z3 and Z4. More embryos arrested and less excellent embryos developed in group Z4 than group Z3. The clinical pregnancy rates resulting from the transfer of single pronuclear score homologous embryo types were similar among groups Z1, Z2 and Z3. Implanta- tion rates of group Z1 were higher (P<0.05) than that of group Z3. These findings suggests that pro- nuclear scoring can predict developmental ability on day 3 and implantation potential. A evaluation that combines the Z-score and day 3 embryo morphology is useful in the determination of the most viable embryos and the number of embryos for transfer.

  8. The correlation between endometrial thickness and outcome of in vitro fertilization and embryo transfer (IVF-ET outcome

    Directory of Open Access Journals (Sweden)

    Al-Rejjal Rafat

    2008-09-01

    Full Text Available Abstract Background To evaluate the relationship between endometrial thickness on day of human chorionic gonadotrophin administration (hCG and pregnancy outcome in a large number of consecutive in vitro fertilization and embryo transfer (IVF-ET cycles. Methods A retrospective cohort study including all patients who had IVF-ET from January 2003–December 2005 conducted at a tertiary center. Results A total of 2464 cycles were analysed. Pregnancy rate (PR was 35.8%. PR increased linearly (r = 0.864 from 29.4% among patients with a lining of less than or equal to 6 mm, to 44.4% among patients with a lining of greater than or equal to 17 mm. ROC showed that endometrial thickness is not a good predictor of PR, so a definite cut-off value could not be established (AUC = 0.55. Conclusion There is a positive linear relationship between the endometrial thickness measured on the day of hCG injection and PR, and is independent of other variables. Hence aiming for a thicker endometrium should be considered.

  9. The HIST1 Locus Escapes Reprogramming in Cloned Bovine Embryos

    Directory of Open Access Journals (Sweden)

    Byungkuk Min

    2016-05-01

    Full Text Available Epigenetic reprogramming is necessary in somatic cell nuclear transfer (SCNT embryos in order to erase the differentiation-associated epigenetic marks of donor cells. However, such epigenetic memories often persist throughout the course of clonal development, thus decreasing cloning efficiency. Here, we explored reprogramming-refractory regions in bovine SCNT blastocyst transcriptomes. We observed that histone genes residing in the 1.5 Mb spanning the cow HIST1 cluster were coordinately downregulated in SCNT blastocysts. In contrast, both the nonhistone genes of this cluster, and histone genes elsewhere remained unaffected. This indicated that the downregulation was specific to HIST1 histone genes. We found that, after trichostatin A treatment, HIST1 histone genes were derepressed, and DNA methylation at their promoters was decreased to the level of in vitro fertilization embryos. Therefore, our results indicate that the reduced expression of HIST1 histone genes is a consequence of poor epigenetic reprogramming in SCNT blastocysts.

  10. A Case of Unicornuate Uterus with Atypical Located Hyperstimulated Ovary after in Vitro Fertilization Pre-Embryo Transfer (IVF-ET

    Directory of Open Access Journals (Sweden)

    Mariya Angelova Angelova

    2015-06-01

    Full Text Available The authors describe a case of a congenital Mullerian anomaly, uterus unicornis with missing right fallopian tube. An in Vitro Fertilization Pre-Embryo Transfer (IVF-ET procedure was done and presently is known that the patient has left fallopian tube and left ovary, two kidneys, and right ovary is missing. No diagnostic laparoscopy and hysteroscopy were done, only hysterosalpingography (HSG before the IVF procedure.  Several days after the follicular puncture of the left ovary the patient was urgently admitted to the hospital for specialized gynaecology in Varna. Transabdominal ultrasonography showed right ovary atypically located immediately next to the liver and with emerging theca-lutein cysts.

  11. N, N-Dimethylglycine decreases oxidative stress and improves in vitro development of bovine embryos.

    Science.gov (United States)

    Takahashi, Toshikiyo; Sasaki, Kouya; Somfai, Tamas; Nagai, Takashi; Manabe, Noboru; Edashige, Keisuke

    2016-04-22

    The antioxidant effect of N, N-dimethylglycine (DMG) on in vitro-produced (IVP) bovine embryos was examined. After in vitro fertilization, presumptive zygotes were cultured with or without 0.1 μM DMG under different oxygen tensions. The percentage of embryos developing to the blastocyst stage was lowest under a 20% oxygen concentration without DMG, and it was significantly increased (P culture medium significantly (P invitro development of IVP bovine embryos by acting as an antioxidant. PMID:26875568

  12. Does transferring three or more embryos make sense for a well-defined population of infertility patients undergoing IVF/ICSI?

    Science.gov (United States)

    Masschaele, T.; Gerris, J.; Vandekerckhove, F.; De Sutter, P.

    2012-01-01

    Recently, there has been a marked increase in the use of Single Embryo Transfer (SET) subsequent to In Vitro Fertilization (IVF) and Intracytoplasmic Sperm Injection (ICSI), with the aim of reducing the risk of multiple gestations. However, critics have stated that by reducing the number of embryos transferred, a group of women with an a priori reduced chance of pregnancy are at particular greater risk of undertreatment. This group includes women who are of a certain age (≥ 40 years) or have already received a number of – failed – IVF attempts. We wanted to study whether the practice of three or more embryos being transferred would be of added value to these patients and whether the strategy of Heavy Load Transfer (HLT) is likely to boost the pregnancy rates to an acceptable level. We performed both a literature study and a retrospective cohort analysis of 7,850 IVF/ICSI cycles of early cleavage stage embryo transfer. Notwithstanding the limitations associated with this approach, we contend that HLT in the group of patients with poor prognosis should be recommended. This article outlines a suggested protocol within the legal framework relevant to Belgium. PMID:24753889

  13. Effects of bone morphogenic protein 4 (BMP4 and its inhibitor, Noggin, on in vitro maturation and culture of bovine preimplantation embryos

    Directory of Open Access Journals (Sweden)

    Fernandez-Martin Rafael

    2011-02-01

    Full Text Available Abstract Background BMP4 is a member of the transforming growth factor beta (TGFbeta superfamily and Noggin is a potent BMP inhibitor that exerts its function by binding to BMPs preventing interactions with its receptors. The aim of this work was to investigate the role of BMP4 and Noggin, on oocytes in vitro maturation (m experiments and embryos in vitro development (c experiments of bovine. Methods For m experiments, COCs were collected from slaughterhouse ovaries and in vitro matured in TCM with 100 ng/ml of either BMP4 or Noggin. After 24 h, the nuclear stage of the oocytes was determined by staining with Hoechst 33342. In addition, RT-qPCR was performed on MII oocytes to study the relative concentration of ZAR1, GDF9, BAX, MATER and HSP70 transcripts. Treated oocytes were submitted to parthenogenic activation (PA or in vitro fertilization (IVF and cultured in CR2. For c experiments, non-treated matured oocytes were submitted to PA or IVF to generate embryos that were exposed to 100 ng/ml of BMP4 or Noggin in CR2 until day nine of culture. Cleavage, blastocyst and hatching rates, expression pattern of the transcription factor Oct-4 in blastocysts and embryo cell number at day two and nine post-activation or fertilization were evaluated. Results We found that Noggin, as BMP4, did not affect oocyte nuclear maturation. Noggin supplementation up-regulated the expression of HSP70 and MATER genes in matured oocytes. Moreover, BMP4 during maturation increased the proportion of Oct-4 positive cells in parthenogenic embryos. On the other hand, when Noggin was added to embryo culture medium, developmental rates of parthenogenic and in vitro fertilized embryos were reduced. However, BMP4 addition decreases the development only for in vitro fertilized embryos. BMP4 and Noggin during culture reduced the proportion of Oct-4-expressing cells. Conclusions Our results show that BMP4 is implicated in bovine oocytes maturation and embryo development. Moreover

  14. Determining the status of non-transferred embryos in Ireland: a conspectus of case law and implications for clinical IVF practice.

    LENUS (Irish Health Repository)

    Sills, Eric Scott

    2009-01-01

    The development of in vitro fertilisation (IVF) as a treatment for human infertilty was among the most controversial medical achievements of the modern era. In Ireland, the fate and status of supranumary (non-transferred) embryos derived from IVF brings challenges both for clinical practice and public health policy because there is no judicial or legislative framework in place to address the medical, scientific, or ethical uncertainties. Complex legal issues exist regarding informed consent and ownership of embryos, particularly the use of non-transferred embryos if a couple separates or divorces. But since case law is only beginning to emerge from outside Ireland and because legislation on IVF and human embryo status is entirely absent here, this matter is poised to raise contractual, constitutional and property law issues at the highest level. Our analysis examines this medico-legal challenge in an Irish context, and summarises key decisions on this issue rendered from other jurisdictions. The contractual issues raised by the Roche case regarding informed consent and the implications the initial judgment may have for future disputes over embryos are also discussed. Our research also considers a putative Constitutional \\'right to procreate\\' and the implications EU law may have for an Irish case concerning the fate of frozen embryos. Since current Medical Council guidelines are insufficient to ensure appropriate regulation of the advanced reproductive technologies in Ireland, the report of the Commission on Assisted Human Reproduction is most likely to influence embryo custody disputes. Public policy requires the establishment and implementation of a more comprehensive legislative framework within which assisted reproductive medical services are offered.

  15. Prospective Randomized Study on the Influence of Myoinositol in PCOS Women Undergoing IVF in the Improvement of Oocyte Quality, Fertilization Rate, and Embryo Quality

    Science.gov (United States)

    Lesoine, Bernd

    2016-01-01

    Polycystic ovarian syndrome (PCOS) is one of the pathological factors involved in the failure of in vitro fertilization (IvF). The aim of the present study was to investigate if the combination of myoinositol + folic acid was able to improve the oocyte quality, the ratio between follicles and retrieved oocytes, the fertilization rate, and the embryo quality in PCOS patients undergoing IvF treatments. 29 patients with PCOS underwent IvF protocols for infertility treatment and were randomized prospectively into two groups. Group A (placebo) with 15 patients and group B (4000 mg myoinositol + 400 μg folic acid per day) with 14 patients. The patients of group B used for two months myoinositol + folic acid before starting the IvF protocol and data were obtained concerning number of follicles, number of oocytes, quality of oocytes, fertilization rates, and embryo quality in both groups. The ratio follicle/retrieved oocyte was better in the myoinositol group (= group B). Out of the 233 oocytes collected in the myoinositol group 136 were fertilized, whereas only 128 out of 300 oocytes in the placebo group were fertilized. More metaphase II and I oocytes were retrieved in relation to the total amount of oocytes in the myoinositol. More embryos of grade I quality were obtained in the myoinositol. The duration of stimulation was 9,7 days (±3,3) in the myoinositol group and 11,2 (±1,8) days in the placebo group and the number of used FSH units was lower in the myoinositol group: 1750 FSH units (mean) versus 1850 units (mean). Our evidence suggests that myoinositol therapy in women with PCOS results in better fertilization rates and a clear trend to a better embryo quality. As the number of retrieved oocytes was smaller in the myoinositol group, the risk of hyper stimulation syndrome can be reduced in these patients. PMID:27635136

  16. Prospective Randomized Study on the Influence of Myoinositol in PCOS Women Undergoing IVF in the Improvement of Oocyte Quality, Fertilization Rate, and Embryo Quality

    Directory of Open Access Journals (Sweden)

    Bernd Lesoine

    2016-01-01

    Full Text Available Polycystic ovarian syndrome (PCOS is one of the pathological factors involved in the failure of in vitro fertilization (IvF. The aim of the present study was to investigate if the combination of myoinositol + folic acid was able to improve the oocyte quality, the ratio between follicles and retrieved oocytes, the fertilization rate, and the embryo quality in PCOS patients undergoing IvF treatments. 29 patients with PCOS underwent IvF protocols for infertility treatment and were randomized prospectively into two groups. Group A (placebo with 15 patients and group B (4000 mg myoinositol + 400 μg folic acid per day with 14 patients. The patients of group B used for two months myoinositol + folic acid before starting the IvF protocol and data were obtained concerning number of follicles, number of oocytes, quality of oocytes, fertilization rates, and embryo quality in both groups. The ratio follicle/retrieved oocyte was better in the myoinositol group (= group B. Out of the 233 oocytes collected in the myoinositol group 136 were fertilized, whereas only 128 out of 300 oocytes in the placebo group were fertilized. More metaphase II and I oocytes were retrieved in relation to the total amount of oocytes in the myoinositol. More embryos of grade I quality were obtained in the myoinositol. The duration of stimulation was 9,7 days (±3,3 in the myoinositol group and 11,2 (±1,8 days in the placebo group and the number of used FSH units was lower in the myoinositol group: 1750 FSH units (mean versus 1850 units (mean. Our evidence suggests that myoinositol therapy in women with PCOS results in better fertilization rates and a clear trend to a better embryo quality. As the number of retrieved oocytes was smaller in the myoinositol group, the risk of hyper stimulation syndrome can be reduced in these patients.

  17. Prospective Randomized Study on the Influence of Myoinositol in PCOS Women Undergoing IVF in the Improvement of Oocyte Quality, Fertilization Rate, and Embryo Quality

    Science.gov (United States)

    Lesoine, Bernd

    2016-01-01

    Polycystic ovarian syndrome (PCOS) is one of the pathological factors involved in the failure of in vitro fertilization (IvF). The aim of the present study was to investigate if the combination of myoinositol + folic acid was able to improve the oocyte quality, the ratio between follicles and retrieved oocytes, the fertilization rate, and the embryo quality in PCOS patients undergoing IvF treatments. 29 patients with PCOS underwent IvF protocols for infertility treatment and were randomized prospectively into two groups. Group A (placebo) with 15 patients and group B (4000 mg myoinositol + 400 μg folic acid per day) with 14 patients. The patients of group B used for two months myoinositol + folic acid before starting the IvF protocol and data were obtained concerning number of follicles, number of oocytes, quality of oocytes, fertilization rates, and embryo quality in both groups. The ratio follicle/retrieved oocyte was better in the myoinositol group (= group B). Out of the 233 oocytes collected in the myoinositol group 136 were fertilized, whereas only 128 out of 300 oocytes in the placebo group were fertilized. More metaphase II and I oocytes were retrieved in relation to the total amount of oocytes in the myoinositol. More embryos of grade I quality were obtained in the myoinositol. The duration of stimulation was 9,7 days (±3,3) in the myoinositol group and 11,2 (±1,8) days in the placebo group and the number of used FSH units was lower in the myoinositol group: 1750 FSH units (mean) versus 1850 units (mean). Our evidence suggests that myoinositol therapy in women with PCOS results in better fertilization rates and a clear trend to a better embryo quality. As the number of retrieved oocytes was smaller in the myoinositol group, the risk of hyper stimulation syndrome can be reduced in these patients.

  18. Prospective Randomized Study on the Influence of Myoinositol in PCOS Women Undergoing IVF in the Improvement of Oocyte Quality, Fertilization Rate, and Embryo Quality.

    Science.gov (United States)

    Lesoine, Bernd; Regidor, Pedro-Antonio

    2016-01-01

    Polycystic ovarian syndrome (PCOS) is one of the pathological factors involved in the failure of in vitro fertilization (IvF). The aim of the present study was to investigate if the combination of myoinositol + folic acid was able to improve the oocyte quality, the ratio between follicles and retrieved oocytes, the fertilization rate, and the embryo quality in PCOS patients undergoing IvF treatments. 29 patients with PCOS underwent IvF protocols for infertility treatment and were randomized prospectively into two groups. Group A (placebo) with 15 patients and group B (4000 mg myoinositol + 400 μg folic acid per day) with 14 patients. The patients of group B used for two months myoinositol + folic acid before starting the IvF protocol and data were obtained concerning number of follicles, number of oocytes, quality of oocytes, fertilization rates, and embryo quality in both groups. The ratio follicle/retrieved oocyte was better in the myoinositol group (= group B). Out of the 233 oocytes collected in the myoinositol group 136 were fertilized, whereas only 128 out of 300 oocytes in the placebo group were fertilized. More metaphase II and I oocytes were retrieved in relation to the total amount of oocytes in the myoinositol. More embryos of grade I quality were obtained in the myoinositol. The duration of stimulation was 9,7 days (±3,3) in the myoinositol group and 11,2 (±1,8) days in the placebo group and the number of used FSH units was lower in the myoinositol group: 1750 FSH units (mean) versus 1850 units (mean). Our evidence suggests that myoinositol therapy in women with PCOS results in better fertilization rates and a clear trend to a better embryo quality. As the number of retrieved oocytes was smaller in the myoinositol group, the risk of hyper stimulation syndrome can be reduced in these patients. PMID:27635136

  19. Stem cells from residual IVF-embryos - Continuation of life justifies isolation.

    NARCIS (Netherlands)

    Bongaerts, G.P.A.; Severijnen, R.S.V.M.

    2007-01-01

    Embryonic stem cells are undifferentiated pluripotent cells that can indefinitely grow in vitro. They are derived from the inner mass of early embryos. Because of their ability to differentiate into all three embryonic germ layers, and finally into specialized somatic cell types, human embryonic ste

  20. Aspects of energetic substrate metabolism of in vitro and in vivo bovine embryos

    International Nuclear Information System (INIS)

    Although the metabolism of early bovine embryos has not been fully elucidated, several publications have addressed this important issue to improve culture conditions for cattle reproductive biotechnologies, with the ultimate goal of producing in vitro embryos similar in quality to those developing in vivo. Here, we review general aspects of bovine embryo metabolism in vitro and in vivo, and discuss the use of metabolic analysis of embryos produced in vitro to assess viability and predict a viable pregnancy after transference to the female tract

  1. Aspects of energetic substrate metabolism of in vitro and in vivo bovine embryos

    Directory of Open Access Journals (Sweden)

    D.K. de Souza

    2015-03-01

    Full Text Available Although the metabolism of early bovine embryos has not been fully elucidated, several publications have addressed this important issue to improve culture conditions for cattle reproductive biotechnologies, with the ultimate goal of producing in vitro embryos similar in quality to those developing in vivo. Here, we review general aspects of bovine embryo metabolism in vitro and in vivo, and discuss the use of metabolic analysis of embryos produced in vitro to assess viability and predict a viable pregnancy after transference to the female tract.

  2. Aspects of energetic substrate metabolism of in vitro and in vivo bovine embryos

    Energy Technology Data Exchange (ETDEWEB)

    Souza, D.K. de [Laboratório de Biotecnologia da Saúde, Faculdade de Medicina, Universidade de Brasília, Brasília, DF (Brazil); Faculdade da Ceilândia, Universidade de Brasília, Brasília, DF (Brazil); Salles, L.P. [Laboratório de Biotecnologia da Saúde, Faculdade de Medicina, Universidade de Brasília, Brasília, DF (Brazil); Departamento de Biologia Molecular, Instituto de Biologia, Universidade de Brasília, Brasília, DF (Brazil); Rosa e Silva, A.A.M. [Laboratório de Biotecnologia da Saúde, Faculdade de Medicina, Universidade de Brasília, Brasília, DF (Brazil)

    2015-01-23

    Although the metabolism of early bovine embryos has not been fully elucidated, several publications have addressed this important issue to improve culture conditions for cattle reproductive biotechnologies, with the ultimate goal of producing in vitro embryos similar in quality to those developing in vivo. Here, we review general aspects of bovine embryo metabolism in vitro and in vivo, and discuss the use of metabolic analysis of embryos produced in vitro to assess viability and predict a viable pregnancy after transference to the female tract.

  3. Relationship between the length of cell cycles, cleavage pattern and developmental competence in bovine embryos generated by in vitro fertilization or parthenogenesis.

    Science.gov (United States)

    Somfai, Tamás; Inaba, Yasushi; Aikawa, Yoshio; Ohtake, Masaki; Kobayashi, Shuji; Konishi, Kazuyuki; Imai, Kei

    2010-04-01

    This study was conducted to study the kinetics of initial cell divisions in relation with the cleavage patterns in viable (with the ability to develop to the blastocyst stage) and non-viable bovine embryos and parthenotes. The kinetics of in vitro development and cleavage patterns were observed by time lapse cinematography. The length of the first and second but not third cell cycle differed significantly between the viable and non-viable embryos after IVF or parthenogenesis. Viable embryos had significantly shorter first and second cell cycles than non-viable ones. The presence of fragments, protrusions and unequally-sized blastomeres was associated with an extended one-cell stage and reduced ability to develop to the blastocyst stage; however, the lengths of the second and third cell cycles were not altered. Oocytes showing direct division from one cell to 3 or 4 blastomeres showed similar developmental ability and embryonic cell numbers to those showing normal division, although, with a high frequency of chromosomal abnormalities. Our results suggest that the differences in the first cell cycles between viable and non-viable embryos were not sperm-related, whereas direct cleavage of 1-cell embryos to 3 or more blastomeres and protrusion formation are related to sperm-driven factors. The length of the first and second cell cycles and the cleavage pattern should be examined simultaneously to predict developmental competence of embryos at early cleavage stages. PMID:20035110

  4. 使用time-lapse筛选早期IVF/ICSI胚胎及其临床结局%Embryo selection in IVF/ICSI cycles using time- lapse microscopy and the clinical outcomes

    Institute of Scientific and Technical Information of China (English)

    陈明颢; 黄军; 钟影; 全松

    2015-01-01

    目的:通过比较使用time-lapse(延迟摄像)和传统形态学方法筛选IVF/ICSI胚胎的临床结局,评价time-lapse用于早期胚胎观察和筛选的价值。方法回顾性分析139个IVF/ICSI周期的资料,根据胚胎的筛选方法,分为time-lapse monitoring组(TLM组)(n=68)和对照组(n=71),比较两组间的βHCG阳性率、临床妊娠率和胚胎着床率,并根据女方年龄、受精方式进行亚组分析。结果 TLM组的βHCG阳性率、临床妊娠率、胚胎着床率分别为:66.2%、61.8%、47.1%;对照组的βHCG阳性率、临床妊娠率、胚胎着床率分别为:47.9%、43.7%、30.3%;TLM组的βHCG阳性率、临床妊娠率、胚胎着床率均高于对照组,且差异均有统计学意义(P30岁)或者是进行IVF周期的患者更能从中获益。%Objective To compare the clinical outcomes of embryos selected using time-lapse microscopy and traditional morphological method in IVF/ICSI cycles and evaluate the clinical value of time-lapse microscopy in early embryo monitoring and selection. Methods We retrospectively analyzed the clinical data of 139 IVF/ICSI cycles with embryo selection based on time-lapse monitoring (TLM group, n=68) and traditional morphological method (control group, n=71). The βHCG-positive rate, clinical pregnancy rate and embryo implantation rate were compared between the 2 groups. Subgroup analysis was performed in view of female patients' age and the fertilization type. Results TheβHCG-positive rate, clinical pregnancy rate and implantation rate were 66.2%, 61.8%and 47.1%in TLM group, significantly higher than those in the control group (47.9%, 43.7%and 30.3%, respectively;P<0.05). Compared with patients below 30 years of age, patients aged between 31 and 35 years benefited more from time-lapse monitoring with improved clinical outcomes. time-lapse monitoring significantly increased theβHCG-positive rate, clinical pregnancy rate and implantation rate for

  5. Efficient delivery of DNA into bovine preimplantation embryos by multiwall carbon nanotubes

    Science.gov (United States)

    Munk, Michele; Ladeira, Luiz O.; Carvalho, Bruno C.; Camargo, Luiz S. A.; Raposo, Nádia R. B.; Serapião, Raquel V.; Quintão, Carolina C. R.; Silva, Saulo R.; Soares, Jaqueline S.; Jorio, Ado; Brandão, Humberto M.

    2016-01-01

    The pellucid zone (PZ) is a protective embryonic cells barrier against chemical, physical or biological substances. This put, usual transfection methods are not efficient for mammal oocytes and embryos as they are exclusively for somatic cells. Carbon nanotubes have emerged as a new method for gene delivery, and they can be an alternative for embryos transfection, however its ability to cross the PZ and mediated gene transfer is unknown. Our data confirm that multiwall carbon nanotubes (MWNTs) can cross the PZ and delivery of pDNA into in vitro-fertilized bovine embryos. The degeneration rate and the expression of genes associated to cell viability were not affected in embryos exposed to MWNTs. Those embryos, however, had lower cell number and higher apoptotic cell index, but this did not impair the embryonic development. This study shows the potential utility of the MWNT for the development of new method for delivery of DNA into bovine embryos. PMID:27642034

  6. Single-embryo transfer reduces clinical pregnancy rates and live births in fresh IVF and Intracytoplasmic Sperm Injection (ICSI cycles: a meta-analysis

    Directory of Open Access Journals (Sweden)

    Oliveira João

    2009-04-01

    Full Text Available Abstract Background It has become an accepted procedure to transfer more than one embryo to the patient to achieve acceptable ongoing pregnancy rates. However, transfers of more than a single embryo increase the probability of establishing a multiple gestation. Single-embryo transfer can minimize twin pregnancies but may also lower live birth rates. This meta-analysis aimed to compare current data on single-embryo versus double-embryo transfer in fresh IVF/ICSI cycles with respect to implantation, ongoing pregnancy and live birth rates. Methods Search strategies included on-line surveys of databases from 1995 to 2008. Data management and analysis were conducted using the Stats Direct statistical software. The fixed-effect model was used for odds ratio (OR. Fixed-effect effectiveness was evaluated by the Mantel Haenszel method. Seven trials fulfilled the inclusion criteria. Results When pooling results under the fixed-effect model, the implantation rate was not significantly different between double-embryo transfer (34.5% and single-embryo transfer group (34.7% (P = 0.96; OR = 0.99, 95% CI 0.78, 1.25. On the other hand, double-embryo transfer produced a statistically significantly higher ongoing clinical pregnancy rate (44.5% than single-embryo transfer (28.3% (P Conclusion Meta-analysis with 95% confidence showed that, despite similar implantation rates, fresh double-embryo transfer had a 1.64 to 2.60 times greater ongoing pregnancy rate and 1.44 to 2.42 times greater live birth rate than single-embryo transfer in a population suitable for ART treatment.

  7. Expression Profile of Genes as Indicators of Developmental Competence and Quality of In Vitro Fertilization and Somatic Cell Nuclear Transfer Bovine Embryos

    Science.gov (United States)

    Monteleone, Melisa Carolina; Mucci, Nicolas; Kaiser, German Gustavo; Brocco, Marcela; Mutto, Adrián

    2014-01-01

    Reproductive biotechnologies such as in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) enable improved reproductive efficiency of animals. However, the birth rate of in vitro-derived embryos still lags behind that of their in vivo counterparts. Thus, it is critical to develop an accurate evaluation and prediction system of embryo competence, both for commercial purposes and for scientific research. Previous works have demonstrated that in vitro culture systems induce alterations in the relative abundance (RA) of diverse transcripts and thus compromise embryo quality. The aim of this work was to analyze the RA of a set of genes involved in cellular stress (heat shock protein 70-kDa, HSP70), endoplasmic reticulum (ER) stress (immunoglobulin heavy chain binding protein, Bip; proteasome subunit β5, PSMB5) and apoptosis (BCL-2 associated X protein, Bax; cysteine aspartate protease-3, Caspase-3) in bovine blastocysts produced by IVF or SCNT and compare it with that of their in vivo counterparts. Poly (A) + mRNA was isolated from three pools of 10 blastocysts per treatment and analyzed by real-time RT-PCR. The RA of three of the stress indicators analyzed (Bax, PSMB5 and Bip) was significantly increased in SCNT embryos as compared with that of in vivo-derived blastocysts. No significant differences were found in the RA of HSP70 and Caspase-3 gene transcripts. This study could potentially complement morphological analyses in the development of an effective and accurate technique for the diagnosis of embryo quality, ultimately aiding to improve the efficiency of assisted reproductive techniques (ART). PMID:25269019

  8. Expression profile of genes as indicators of developmental competence and quality of in vitro fertilization and somatic cell nuclear transfer bovine embryos.

    Directory of Open Access Journals (Sweden)

    Maria Jesús Cánepa

    Full Text Available Reproductive biotechnologies such as in vitro fertilization (IVF and somatic cell nuclear transfer (SCNT enable improved reproductive efficiency of animals. However, the birth rate of in vitro-derived embryos still lags behind that of their in vivo counterparts. Thus, it is critical to develop an accurate evaluation and prediction system of embryo competence, both for commercial purposes and for scientific research. Previous works have demonstrated that in vitro culture systems induce alterations in the relative abundance (RA of diverse transcripts and thus compromise embryo quality. The aim of this work was to analyze the RA of a set of genes involved in cellular stress (heat shock protein 70-kDa, HSP70, endoplasmic reticulum (ER stress (immunoglobulin heavy chain binding protein, Bip; proteasome subunit β5, PSMB5 and apoptosis (BCL-2 associated X protein, Bax; cysteine aspartate protease-3, Caspase-3 in bovine blastocysts produced by IVF or SCNT and compare it with that of their in vivo counterparts. Poly (A + mRNA was isolated from three pools of 10 blastocysts per treatment and analyzed by real-time RT-PCR. The RA of three of the stress indicators analyzed (Bax, PSMB5 and Bip was significantly increased in SCNT embryos as compared with that of in vivo-derived blastocysts. No significant differences were found in the RA of HSP70 and Caspase-3 gene transcripts. This study could potentially complement morphological analyses in the development of an effective and accurate technique for the diagnosis of embryo quality, ultimately aiding to improve the efficiency of assisted reproductive techniques (ART.

  9. Associations between Individual and Combined Polymorphisms of the TNF and VEGF Genes and the Embryo Implantation Rate in Patients Undergoing In Vitro Fertilization (IVF Programs.

    Directory of Open Access Journals (Sweden)

    Radia Boudjenah

    Full Text Available A multiple pregnancy is now considered to be the most common adverse outcome associated with in vitro fertilization (IVF. As a consequence, the identification of women with the best chances of embryo implantation is a challenge in IVF program, in which the objective is to offer elective single-embryo transfer (eSET without decreasing the pregnancy rate. To date, a range of hormonal and clinical parameters have been used to optimize eSET but none have significant predictive value. This variability could be due to genetic predispositions related to single-nucleotide polymorphisms (SNPs. Here, we assessed the individual and combined impacts of thirteen SNPs that reportedly influence the outcome of in vitro fertilisation (IVF on the embryo implantation rate for patients undergoing intracytoplasmic sperm injection program (ICSI.A 13 gene polymorphisms: FSHR(Asn680Ser, p53(Arg72Pro, AMH(Ile49Ser, ESR2(+1730G>A, ESR1(-397T>C, BMP15(-9C>G, MTHFR1(677C>T, MTHFR2(1298A>C, HLA-G(-725C>G, VEGF(+405G>C, TNFα(-308A>G, AMHR(-482A>G, PAI-1(4G/5G, multiplex PCR assay was designed to genotype women undergoing ICSI program. We analyzed the total patients population (n = 428 and a subgroup with homogeneous characteristics (n = 112.Only the VEGF(+405G>C and TNFα(-308A>G polymorphisms impacted fertilization, embryo implantation and pregnancy rates. Moreover, the combined VEGF+405.GG and TNFα-308.AG or AA genotype occurred significantly more frequently in women with high implantation potential. In contrast, the VEGF+405.CC and TNFα-308.GG combination was associated with a low implantation rate.We identified associations between VEGF(+405G>C and TNFα(-308A>G polymorphisms (when considered singly or as combinations and the embryo implantation rate. These associations may be predictive of embryo implantation and could help to define populations in which elective single-embryo transfer should be recommended (or, conversely, ruled out. However, the mechanism

  10. Porcine Pluripotent Stem Cells Derived from IVF Embryos Contribute to Chimeric Development In Vivo.

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    Binghua Xue

    Full Text Available Although the pig is considered an important model of human disease and an ideal animal for the preclinical testing of cell transplantation, the utility of this model has been hampered by a lack of genuine porcine embryonic stem cells. Here, we derived a porcine pluripotent stem cell (pPSC line from day 5.5 blastocysts in a newly developed culture system based on MXV medium and a 5% oxygen atmosphere. The pPSCs had been passaged more than 75 times over two years, and the morphology of the colony was similar to that of human embryonic stem cells. Characterization and assessment showed that the pPSCs were alkaline phosphatase (AKP positive, possessed normal karyotypes and expressed classic pluripotent markers, including OCT4, SOX2 and NANOG. In vitro differentiation through embryonic body formation and in vivo differentiation via teratoma formation in nude mice demonstrated that the pPSCs could differentiate into cells of the three germ layers. The pPSCs transfected with fuw-DsRed (pPSC-FDs could be passaged with a stable expression of both DsRed and pluripotent markers. Notably, when pPSC-FDs were used as donor cells for somatic nuclear transfer, 11.52% of the reconstructed embryos developed into blastocysts, which was not significantly different from that of the reconstructed embryos derived from porcine embryonic fibroblasts. When pPSC-FDs were injected into day 4.5 blastocysts, they became involved in the in vitro embryonic development and contributed to the viscera of foetuses at day 50 of pregnancy as well as the developed placenta after the chimeric blastocysts were transferred into recipients. These findings indicated that the pPSCs were porcine pluripotent cells; that this would be a useful cell line for porcine genetic engineering and a valuable cell line for clarifying the molecular mechanism of pluripotency regulation in pigs.

  11. Porcine Pluripotent Stem Cells Derived from IVF Embryos Contribute to Chimeric Development In Vivo

    Science.gov (United States)

    Xue, Binghua; Li, Yan; He, Yilong; Wei, Renyue; Sun, Ruizhen; Yin, Zhi; Bou, Gerelchimeg; Liu, Zhonghua

    2016-01-01

    Although the pig is considered an important model of human disease and an ideal animal for the preclinical testing of cell transplantation, the utility of this model has been hampered by a lack of genuine porcine embryonic stem cells. Here, we derived a porcine pluripotent stem cell (pPSC) line from day 5.5 blastocysts in a newly developed culture system based on MXV medium and a 5% oxygen atmosphere. The pPSCs had been passaged more than 75 times over two years, and the morphology of the colony was similar to that of human embryonic stem cells. Characterization and assessment showed that the pPSCs were alkaline phosphatase (AKP) positive, possessed normal karyotypes and expressed classic pluripotent markers, including OCT4, SOX2 and NANOG. In vitro differentiation through embryonic body formation and in vivo differentiation via teratoma formation in nude mice demonstrated that the pPSCs could differentiate into cells of the three germ layers. The pPSCs transfected with fuw-DsRed (pPSC-FDs) could be passaged with a stable expression of both DsRed and pluripotent markers. Notably, when pPSC-FDs were used as donor cells for somatic nuclear transfer, 11.52% of the reconstructed embryos developed into blastocysts, which was not significantly different from that of the reconstructed embryos derived from porcine embryonic fibroblasts. When pPSC-FDs were injected into day 4.5 blastocysts, they became involved in the in vitro embryonic development and contributed to the viscera of foetuses at day 50 of pregnancy as well as the developed placenta after the chimeric blastocysts were transferred into recipients. These findings indicated that the pPSCs were porcine pluripotent cells; that this would be a useful cell line for porcine genetic engineering and a valuable cell line for clarifying the molecular mechanism of pluripotency regulation in pigs. PMID:26991423

  12. Vitrification vs. slow cooling protocol using embryos cryopreserved in the 5th or 6th day after oocyte retrieval and IVF outcomes.

    Directory of Open Access Journals (Sweden)

    Joanna Matysiak

    2010-06-01

    Full Text Available Modifying cryopreservation protocols may be seen as a way to simplify cryobanking procedure and increase satisfying outcomes. The aim of the study was to evaluate the influence of slow cooling protocol and vitrification on IVF outcomes using embryos preserved in the 5th or 6th day after oocyte retrieval. The study compared 2 groups of human embryos underwent slow cooling protocol (n=189 and vitrification (n=58. All embryos were cryopreserved in the 5th or 6th day after oocyte retrieval. Pre- and postfreezing embryo evaluation was performed in 2 or 3 steps scale, respectively. The study evaluates the effectiveness of two freezing methods and influence of the freezing day, pre- and postfreezing embryo grading on clinical pregnancy rate. Study showed higher pregnancy rate after vitrification (50.4% than slow cooling protocol (25.9%. Significantly higher pregnancy rate was observed, when embryo preserved in the 5th day after oocyte retrieval (50.3% than in the 6th day (22.7%. Postfreezing embryos evaluation showed that high quality blastocysts gave nearly four times better pregnancy outcomes than the ones evaluated as poor quality, and three times better than the ones evaluated as moderate. Prospective trials are needed to evaluate pregnancy and neonatal outcomes after vitrification. The number of controlled studies concerning vitrification has not been large enough, yet.

  13. Bed rest following embryo transfer might negatively affect the outcome of IVF/ICSI: a systematic review and meta-analysis.

    Science.gov (United States)

    Craciunas, Laurentiu; Tsampras, Nikolaos

    2016-04-01

    The majority of patients undergoing in vitro fertilization (IVF) and intracytoplasmatic sperm injection (ICSI) treatment will reach the stage of embryo transfer (ET), but only a small proportion of transferred embryos implant. Bed rest following ET has been recommended as a way to prevent embryo expulsion by gravity. We performed a systematic review and meta-analysis of randomized controlled trials (RCTs) published prior to May 2014 reporting the effect of bed rest following ET, and irrespective of language, country of origin, blinding or sample size. Four RCTs, including 757 women met the inclusion criteria. Bed rest following ET did not improve clinical pregnancy and live birth rates, but reduced the implantation rate. The quality of the trials included was moderate because of attrition bias and possible reporting bias. The findings of this systematic review and meta-analysis are concordant with previously published literature and suggest that bed rest is not beneficial following ET. Moreover, it might negatively affect the outcome of IVF/ICSI cycles via stress/anxiety mechanisms. PMID:26986834

  14. Bed rest following embryo transfer might negatively affect the outcome of IVF/ICSI: a systematic review and meta-analysis.

    Science.gov (United States)

    Craciunas, Laurentiu; Tsampras, Nikolaos

    2016-04-01

    The majority of patients undergoing in vitro fertilization (IVF) and intracytoplasmatic sperm injection (ICSI) treatment will reach the stage of embryo transfer (ET), but only a small proportion of transferred embryos implant. Bed rest following ET has been recommended as a way to prevent embryo expulsion by gravity. We performed a systematic review and meta-analysis of randomized controlled trials (RCTs) published prior to May 2014 reporting the effect of bed rest following ET, and irrespective of language, country of origin, blinding or sample size. Four RCTs, including 757 women met the inclusion criteria. Bed rest following ET did not improve clinical pregnancy and live birth rates, but reduced the implantation rate. The quality of the trials included was moderate because of attrition bias and possible reporting bias. The findings of this systematic review and meta-analysis are concordant with previously published literature and suggest that bed rest is not beneficial following ET. Moreover, it might negatively affect the outcome of IVF/ICSI cycles via stress/anxiety mechanisms.

  15. Green tea polyphenols added to IVM and IVC media affect transcript abundance, apoptosis, and pregnancy rates in bovine embryos.

    Science.gov (United States)

    Wang, Zhengguang; Fu, Chunquan; Yu, Songdong

    2013-01-01

    Three experiments were conducted to examine the effects of green tea polyphenols (GTP) during IVM and IVC on apoptosis and relative transcript abundance (RA) of three genes controlling antioxidant enzymes, as well as subsequent pregnancy rates. In experiment 1, oocytes were matured in the presence of 0, 10, 15, or 25 μM GTP for 24 hours. The GTP dose applied to IVM medium was followed by the same dose supplemented to IVC medium, so oocytes and embryos of a given group were cultured in similar conditions. This resulted in a total of four groups (three experimental groups and the control). After IVF, presumptive zygotes were cultured in medium containing 0 to 25 μM GTP for 8 days. The addition of 15 μM GTP during IVM and IVC increased RA of SOD1, CAT, and GPX genes in blastocysts compared with the control (P < 0.05). Increase in GTP doses from 15 to 25 μM did not further increase the transcript level. In experiment 2, effects of GTP doses on apoptosis were investigated in bovine blastocysts. Two of the applied GTP doses (10 and 15 μM) decreased the apoptotic index (AI) in blastocysts (7.4% and 6.2% respectively) compared with the control (9.3%; P < 0.05). However, the highest GTP dose used (25 μM) caused an increase in AI compared with a dose of 15 μM (P < 0.05). Considering the results of experiment 1 and 2, the effects of 15 μM GTP treatment during IVM and IVC on pregnancy rate was evaluated after embryo transfer in experiment 3. Cows receiving embryos treated with 15 μM GTP had higher pregnancy rates on Day 30 (34.8% vs. 28.6%) and Day 60 (34.8% vs. 23.9%) than those receiving control embryos (P < 0.05). In conclusion, addition of 15 μM GTP during IVM and IVC improved pregnancy rates; this improvement seemed to be associated with the increase of RA of antioxidant enzyme genes and the decrease in AI in bovine blastocysts.

  16. Viability of bovine demi embryo after splitting of fresh and frozen thawed embryo derived from in vitro embryo production

    Directory of Open Access Journals (Sweden)

    M Imron

    2007-06-01

    Full Text Available In vivo embryo production was limited by number of donor, wide variability respond due to superovulation program and also immunoactifity of superovulation hormone (FSH. Splitting technology could be an alternative to increase the number of transferrable embryos into recipien cows. Splitting is done with cutting embryo becoming two equal pieces (called demi embrio base on ICM orientation. The objective of this research was to determine the viability of demi embryo obtained from embryo splitting of fresh and frozen thawed embryo. The results showed that demi embryos which performed blastocoel reexpansion 3 hours after embryo splitting using fresh and frozen thawed embryos were 76.9 and 76.2% respectively. Base on existention of inner cell mass (ICM, the number of demi embryos developed with ICM from fresh and frozen thawed embryos were not significantly different (90.6 and 85.7% respectively. The cell number of demi embryo from fresh embryos splitting was not different compared with those from frozen thawed embryos (36.1 and 35.9 respectively. These finding indicated that embryo splitting can be applied to frozen thawed embryos with certain condition as well as fresh embryos.

  17. Lysophosphatidic Acid Signaling in Late Cleavage and Blastocyst Stage Bovine Embryos

    OpenAIRE

    Ana Catarina Torres; Dorota Boruszewska; Mariana Batista; Ilona Kowalczyk-Zieba; Patricia Diniz; Emilia Sinderewicz; Jean Sebastian Saulnier-Blache; Izabela Woclawek-Potocka; Luis Lopes-da-Costa

    2014-01-01

    Lysophosphatidic acid (LPA) is a known cell signaling lipid mediator in reproductive tissues. In the cow, LPA is involved in luteal and early pregnancy maintenance. Here, we evaluated the presence and role of LPA in bovine early embryonic development. In relevant aspects, bovine embryos reflect more closely the scenario occurring in human embryos than the mouse model. Transcription of mRNA and protein expression of enzymes involved in LPA synthesis (ATX and cPLA2) and of LPA receptors (LPAR1–...

  18. Effect of oocyte quality and activation protocols on bovine embryo development following intracytoplasmic sperm injection

    OpenAIRE

    KORKMAZ, Ömer; KÜPLÜLÜ, Şükrü; AĞCA, Yüksel; POLAT, İbrahim Mert

    2013-01-01

    The purpose of this study was to investigate the effects of oocyte quality and activation protocols on the in vitro developmental competence of bovine embryos after intracytoplasmic sperm injection (ICSI). Bovine oocytes were grouped as being of excellent, good, and poor quality. All of the oocytes were activated using a calcium ionophore only, ethanol only, and 6-dimethylaminopurine (6-DMAP) following calcium ionophore. For the excellent quality oocytes, cleavage rates after ICSI were 70% in...

  19. Influence of antral follicle size on oocyte characteristics and embryo development in the bovine

    DEFF Research Database (Denmark)

    Lequarre, Anne Sophie; Vigneron, Céline; Ribaucour, Fabrice;

    2005-01-01

    The developmental competence of bovine oocytes isolated from antral follicles of different sizes was assessed in three European laboratories (Belgium, UCL; Denmark, DIAS; France, INRA). Using the same protocol for in vitro production of embryos, the oocytes isolated from follicles with a diameter...

  20. Comparison of the efficacy of conventional slow freezing and rapid cryopreservation methods for bovine embryos

    NARCIS (Netherlands)

    Wagtendonk-de Leeuw, van A.M.; den Daas, J.H.; Kruip, T.A.; Rail, W.F.

    1995-01-01

    Day 7 bovine morulae and early blastocysts were randomly assigned to one of four cryopreservation methods: (i) a modified conventional controlled slow freezing and stepwise dilution after thawing; and three methods which enable direct transfer of the embryo into the recipient upon thawing: (ii) conv

  1. Alternative approaches in IVF

    OpenAIRE

    Fauser, Bart; Bouchard, P.; Coelingh Bennink, H.J.; Collins, John; Devroey, P.; Evers, Johannes; Van Steirteghem, A

    2002-01-01

    textabstractVarious new developments in clinical and basic science which may impact on IVF in the near or distant future will be discussed in this review. These key areas include the regulation of early follicle development and the extended in-vitro culture of oocytes and embryos. Moreover, alternative compounds and ovarian stimulation protocols will be discussed, along with highlights in the development of the cryopreservation of excess oocytes or embryos. Finally, the health economics of IV...

  2. Alternative approaches in IVF

    NARCIS (Netherlands)

    B.C.J.M. Fauser (Bart); P. Bouchard; H.J. Coelingh Bennink; J.A. Collins (John); P. Devroey; J.L.H. Evers (Johannes); A. van Steirteghem

    2002-01-01

    textabstractVarious new developments in clinical and basic science which may impact on IVF in the near or distant future will be discussed in this review. These key areas include the regulation of early follicle development and the extended in-vitro culture of oocytes and embryos.

  3. Raman-based noninvasive metabolic profile evaluation of in vitro bovine embryos

    Science.gov (United States)

    dos Santos, Érika Cristina; Martinho, Herculano; Annes, Kelly; da Silva, Thais; Soares, Carlos Alexandre; Leite, Roberta Ferreira; Milazzotto, Marcella Pecora

    2016-07-01

    The timing of the first embryonic cell divisions may predict the ability of an embryo to establish pregnancy. Similarly, metabolic profiles may be markers of embryonic viability. However, in bovine, data about the metabolomics profile of these embryos are still not available. In the present work, we describe Raman-based metabolomic profiles of culture media of bovine embryos with different developmental kinetics (fast x slow) throughout the in vitro culture. The principal component analysis enabled us to classify embryos with different developmental kinetics since they presented specific spectroscopic profiles for each evaluated time point. We noticed that bands at 1076 cm-1 (lipids), 1300 cm-1 (Amide III), and 2719 cm-1 (DNA nitrogen bases) gave the most relevant spectral features, enabling the separation between fast and slow groups. Bands at 1001 cm-1 (phenylalanine) and 2892 cm-1 (methylene group of the polymethylene chain) presented specific patterns related to embryonic stage and can be considered as biomarkers of embryonic development by Raman spectroscopy. The culture media analysis by Raman spectroscopy proved to be a simple and sensitive technique that can be applied with high efficiency to characterize the profiles of in vitro produced bovine embryos with different development kinetics and different stages of development.

  4. Developmental stage on day-5 and fragmentation rate on day-3 can influence the implantation potential of top-quality blastocysts in IVF cycles with single embryo transfer

    Directory of Open Access Journals (Sweden)

    Devroey Paul

    2007-01-01

    Full Text Available Abstract Background In IVF-ICSI cycles with single embryo transfer (SET, embryo selection for transfer is of crucial importance. The present study aimed to define which embryo parameters might be related to the implantation potential of advanced blastocysts. Methods Overall, in 203 cycles with SET, developmental characteristics of 93 implanted (group A and 110 non-implanted (group B advanced blastocysts of good quality were compared. The following developmental parameters were assessed in the two groups: normal fertilization, developmental stage on day 5, number of blastomeres on day 2 and on day 3, fragmentation rate on day 3, compaction on day 4 and cleavage pattern on day 2 and day 3. Results Expanded blastocysts compared to full blastocysts have higher implantation potential (56.5% vs. 29.3%, p 10–50% fragments on day 3 showed a significant lower implantation (29.7% than those with ≤ 10%fragments (49.4%, P = 0.03. All the other parameters analysed were comparable for the two groups. Conclusion Developmental stage on day 5 and fragmentation rate on day 3 were related to the implantation potential of advanced blastocysts and should also be taken into account in the selection of the best advanced blastocyst for transfer.

  5. Embryo-maternal interactions leading to embryonic development and survival in the bovine : role of progesterone and prostaglandins

    OpenAIRE

    Torres, Ana Catarina Belejo Mora

    2012-01-01

    Tese de Doutoramento em Ciências Veterinárias. Especialidade de Clínica The objectives of this thesis were to evaluate steroidogenic and prostanoid embryo-maternal interactions leading to embryonic development and survival in cattle, and to evaluate therapeutic strategies at embryo transfer (ET) designed to enhance embryo survival. In vitro experiments (three experimental chapters) - bovine early (Day 7) embryos i) had transcription of genes coding for enzymes progesterone (P4)...

  6. Morphokinetic-related response to stress in individually cultured bovine embryos.

    Science.gov (United States)

    Silva, T; Santos, E C; Annes, K; Soares, C A; Leite, R F; Lima, C B; Milazzotto, M P

    2016-09-15

    The kinetics of in vitro-produced (IVP) bovine embryos is related to embryo viability, metabolism, and epigenetic patterns. Therefore, we believe that embryos with different speeds of development also respond differently to stress. In the present study, we performed global metabolic analysis (matrix-assisted laser desorption ionization time of flight mass spectrometry [MALDI-TOF]) of culture media, characterized apoptotic events (Terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL] and caspase quantitation), and quantified transcript abundance of stress-related gene (real-time quantitative polymerase chain reaction [qRT-PCR]) in IVP bovine embryos with different developmental kinetics to investigate possible markers of stress response. For this purpose, embryos were considered "fast" if they presented four or more cells at 40 hours post insemination (hpi). Embryos presenting two cells at this time were classified as "slow". Evaluations were performed at 40 hpi, 112 hpi, and 186 hpi. Metabolome analysis revealed several metabolites differentially represented between groups at all time points related with energy, lipid and amino acids metabolism, and stress response. There was no difference in TUNEL positive cells between groups in any of the time points analyzed. Nevertheless, at 112 hpi, classified as a critical phase because of the genome activation, the amount of caspase 3 and 7 and total caspase were higher in slow when compared to fast group. Transcript abundance analysis of candidate genes (GRP78, HSP60, SOD1, and MORF4L2) was also different among groups. In conclusion, IVP bovine embryos of different development speeds respond differentially to the environmental stress leading to different metabolome patterns and apoptosis activation throughout the culture.

  7. Morphokinetic-related response to stress in individually cultured bovine embryos.

    Science.gov (United States)

    Silva, T; Santos, E C; Annes, K; Soares, C A; Leite, R F; Lima, C B; Milazzotto, M P

    2016-09-15

    The kinetics of in vitro-produced (IVP) bovine embryos is related to embryo viability, metabolism, and epigenetic patterns. Therefore, we believe that embryos with different speeds of development also respond differently to stress. In the present study, we performed global metabolic analysis (matrix-assisted laser desorption ionization time of flight mass spectrometry [MALDI-TOF]) of culture media, characterized apoptotic events (Terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL] and caspase quantitation), and quantified transcript abundance of stress-related gene (real-time quantitative polymerase chain reaction [qRT-PCR]) in IVP bovine embryos with different developmental kinetics to investigate possible markers of stress response. For this purpose, embryos were considered "fast" if they presented four or more cells at 40 hours post insemination (hpi). Embryos presenting two cells at this time were classified as "slow". Evaluations were performed at 40 hpi, 112 hpi, and 186 hpi. Metabolome analysis revealed several metabolites differentially represented between groups at all time points related with energy, lipid and amino acids metabolism, and stress response. There was no difference in TUNEL positive cells between groups in any of the time points analyzed. Nevertheless, at 112 hpi, classified as a critical phase because of the genome activation, the amount of caspase 3 and 7 and total caspase were higher in slow when compared to fast group. Transcript abundance analysis of candidate genes (GRP78, HSP60, SOD1, and MORF4L2) was also different among groups. In conclusion, IVP bovine embryos of different development speeds respond differentially to the environmental stress leading to different metabolome patterns and apoptosis activation throughout the culture. PMID:27298151

  8. Factors that affect the reproductive efficiency of the recipient within a bovine embryo transfer program

    Directory of Open Access Journals (Sweden)

    Arturo Duica A.

    2007-12-01

    Full Text Available The embryo transfer is a biotechnological technique that allows increasing the descendant of animals with high genetic value. The positive results, represented in pregnancy after the application of this technique, are affected by some factors that are inherent to the donor, the embryo, the technique, and the recipients which receive a strange embryo in the uterus allowing pregnancy. This review describes some factors affecting the reproductive efficiency of the recipients of bovine embryos within a program of embryo transfer. Its important to evaluate the parameters in this kind of recipients, as race, age, physiological status, health status, weight, reproductive tract integrity and management, and also too monitoring the ovarian structures while the estrus synchronization, and within previous and posterior stages in embryo transfer procedure. Therefore an optimum follicular development will be determinant to corpus luteum formation which generates enough serum progesterone concentrations to offer a right uterine environment allowing the optimum embryo development. Controlling the factors that affect the efficiency of the embryo transfer, it will obtain an increasing of positive results represented in pregnancies and births of individuals come from animals with high genetic value.

  9. Effect of the bovine oviductal fluid on in vitro fertilization, development and gene expression of in vitro-produced bovine blastocysts.

    Science.gov (United States)

    Cebrian-Serrano, A; Salvador, I; García-Roselló, E; Pericuesta, E; Pérez-Cerezales, S; Gutierrez-Adán, A; Coy, P; Silvestre, M A

    2013-04-01

    Oviductal microenvironment generally provides better conditions for early embryo development than the conventional in vitro system. In an attempt to simulate the oviduct conditions or the main potentially influencing factors, the effect was studied of a bovine oviductal fluid (bOF) treatment applied prior to IVF on (i) IVF parameters, (ii) cleavage rate, (iii) blastocyst yield and (iv) blastocyst quality. Embryo quality was assessed by morphological embryo quality and relative transcript abundance of several developmental genes in bovine blastocysts. Furthermore, to study the effect of bOF without the male effect and zona-sperm interaction, artificially activated metaphase II oocytes were also treated with bOF. In vitro-matured bovine oocytes from abattoir ovaries were treated or untreated with bOF for 30 min and then washed prior to IVF or activation. Subsequently, in vitro-fertilized and parthenogenetic embryos were in vitro cultured for 7 to 8 days. The bOF treatment had no effect on fertilization parameters, cleavage, blastocyst rates both on parthenogenetic and IVF bovine embryos and neither on morphological quality of IVF blastocysts. G6PD and SOD2 genes from IVF blastocysts showed significant changes in their expression after a bOF treatment. Significant differences were found for the expression of SCL2A1, GPX1, BAX, AKR1B1 and PLAC8 genes between excellent or good blastocysts (Grade 1) and fair blastocysts (Grade 2). To our knowledge, this is the first study that evaluates the effect of bOF oocyte treatment on fertilization parameters, development and quality of bovine embryos. PMID:22908847

  10. 对废弃胚胎行囊胚培养在体外受精-胚胎移植中的意义%Potential use of discarded embryos for in vitro fertilization and embryo transfer (IVF-ET)

    Institute of Scientific and Technical Information of China (English)

    王荣; 费小阳; 王丽卿; 邹立波

    2014-01-01

    Objective To explore the potential use of discarded embryos for in vitro fertilization- embryo transfer (IVF- ET). Methods The discarded embryos from d3 were cultured into blastula by the sequential culture method. Results A total of 1962 discarded embryos from 782 patients were cultured, in which 376 blastocysts were formed with a blastocyst rate of 19.2%. The blastocyst formation rates were significantly higher from 1PN, 0PN and>6 cells embryos. The clinical pregnancy rate in women with blastosphere formed was significantly higher than those without blastosphere formed. Sixty four blastocysts were formed for 2PN,10 of which were transferred while the others were frozen. The clinical pregnancy rate of blastocyst transfer was 66.7%. Thir-teen women got pregnant in 24 frozen- thawed blastocyst transfer cycles. A total of 18 healthy babies were born from blastocyst transfer. Conclusion The discarded embryos have different development potential.They can develop into biastospheres. Blas-tocyst from appropriate discarded embryos can improve the utilization rate of embryos.%目的:探讨对不可移植的废弃胚胎行囊胚培养在体外受精-胚胎移植(IVF- ET)中的意义。方法通过囊胚序贯培养法将d3废弃胚胎培养至囊胚期。比较不同来源胚胎及d3胚胎的卵裂球数与囊胚形成的关系;利用废弃胚胎囊胚形成情况对体外受精妊娠结局进行预测;并将获得的可移植囊胚行胚胎移植及冷冻。结果共收集782例患者的1962个废弃胚胎,经序贯培养,形成囊胚376枚(19.2%);1PN胚胎、0PN胚胎、d 3卵裂球数>6的胚胎囊胚形成率较高(均P<0.05);废弃胚胎中有囊胚形成者的临床妊娠率明显高于无囊胚形成者(P<0.05);由2PN胚胎形成囊胚共64枚,其中10枚囊胚行移植,剩余囊胚行玻璃化冷冻保存,囊胚移植后临床妊娠率66.7%。解冻移植囊胚24个周期,妊娠周期13个,妊娠率54.2%。囊

  11. Coagulansin-A has beneficial effects on the development of bovine embryos in vitro via HSP70 induction

    OpenAIRE

    Khan, Imran; Lee, Kyeong-Lim; Fakruzzaman, Md.; Song, Seok-Hwan; Ihsan-ul-Haq,; Mirza, Bushra; Yan, Chang Guo; Kong, Il-Keun

    2016-01-01

    Treatment with the steroidal lactone, coagulansin-A, improves bovine oocyte maturation and embryo development in vitro by inducing heat shock protein 70 (HSP70), which reduces the levels of reactive oxygen species (ROS), DNA damage and inflammation.

  12. Expression of antiapoptosis gene survivin in luteinized ovarian granulosa cells of women undergoing IVF or ICSI and embryo transfer: clinical correlations

    Directory of Open Access Journals (Sweden)

    Varras Michail

    2012-09-01

    Full Text Available Abstract Background The purpose of the study was to determine the incidence of survivin gene expression in human granulosa cells during ovarian stimulation in Greek women with normal FSH levels, undergoing IVF or ICSI and to discover any correlation between levels of gene expression and clinical parameters, efficacy of ovulation or outcomes of assisted reproduction. Methods Twenty nine women underwent ovulation induction for IVF or ICSI and ET with standard GnRH analogue-recombinant FSH protocol. Infertility causes were male and tubal factor. Cumulus–mature oocyte complexes were denuded and the granulosa cells were analyzed for each patient separately using quantitative reverse transcription polymerase chain reaction analysis for survivin gene expression with internal standard the ABL gene. Results The ABL and survivin mRNA were detected in granulosa cells in 93.1%. The expression levels of survivin were significantly lower in normal women (male infertility factor compared to women with tubal infertility factor (p = 0.007. There was no additional statistically significant correlation between levels of survivin expression and estradiol levels or dosage of FSH for ovulation induction or number of dominant follicles aspirated or number of retrieved oocytes or embryo grade or clinical pregnancy rates respectively. Conclusions High levels of survivin mRNA expression in luteinized granulosa cells in cases with tubal infertility seem to protect ovaries from follicular apoptosis. A subpopulation of patients with low levels of survivin mRNA in granulosa cells might benefit with ICSI treatment to bypass possible natural barriers of sperm-oocyte interactions.

  13. Proteomic analysis of the early bovine yolk sac fluid and cells from the day 13 ovoid and elongated preimplantation embryos

    DEFF Research Database (Denmark)

    Jensen, Pernille L; Beck, Hans Christian; Petersen, Tonny S;

    2014-01-01

    The bovine blastocyst hatches 8 to 9 days after fertilization, and this is followed by several days of preimplantation development during which the embryo transforms from a spherical over an ovoid to an elongated shape. As the spherical embryo enlarges, the cells of the inner cell mass...... slightly later stage cell differentiation in the developing bovine embryo. In vitro-produced Day 6 embryos were transferred into a recipient heifer and after 7 days of further in vivo culture, ovoid and elongated Day 13 embryos were recovered by flushing both uterine horns after slaughter. The primitive YS...... differentiate into the hypoblast and epiblast, which remain surrounded by the trophectoderm. The formation of the hypoblast epithelium is also accompanied by a change in the fluid within the embryo, i.e., the blastocoel fluid gradually alters to become the primitive yolk sac (YS) fluid. Our previous research...

  14. Direct and Osmolarity-Dependent Effects of Glycine on Preimplantation Bovine Embryos.

    Science.gov (United States)

    Herrick, Jason R; Lyons, Sarah M; Greene, Alison F; Broeckling, Corey D; Schoolcraft, William B; Krisher, Rebecca L

    2016-01-01

    Concentrations of glycine (Gly) in embryo culture media are often lower (~0.1 mM) than those in oviductal or uterine fluids (≥1.2 mM). The objective of this study was to determine direct and osmolarity-dependent effects of physiological concentrations of Gly on blastocyst formation and hatching, cell allocation to the trophectoderm (TE) and inner cell mass (ICM), and metabolic activity of bovine embryos. In experiment 1, zygotes were cultured with 100 or 120 mM NaCl and 0 or 1 mM Gly for the first 72 h of culture. Blastocyst formation and hatching were improved (Pcultured with 100 compared to 120 mM NaCl. Inclusion of 1 mM Gly improved (Pcultured with 120 mM NaCl, suggesting bovine embryos can utilize Gly as an osmolyte. In experiment 2, embryos were cultured with 0.1, 1.1, 2.1, or 4.1 mM Gly (100 mM NaCl) for the final 96 h of culture. Blastocyst development was not affected (P>0.05) by Gly, but hatching (0.1 mM Gly, 18.2%) was improved (Pcultured with 1.1 (31.4%) or 2.1 (29.4%) mM Gly. Blastocyst, TE, and ICM cell numbers were not affected (P>0.05) by Gly in either experiment. Blastocysts produced alanine, glutamine, pyruvate, and urea and consumed aspartate, but this metabolic profile was not affected (P>0.05) by Gly. In conclusion, Gly (1.0 mM) improves the development of both early and late stage embryos, but beneficial effects are more pronounced for early embryos exposed to elevated osmolarity. PMID:27459477

  15. Comparison of transcriptomic landscapes of bovine embryos using RNA-Seq

    Directory of Open Access Journals (Sweden)

    Khatib Hasan

    2010-12-01

    Full Text Available Abstract Background Advances in sequencing technologies have opened a new era of high throughput investigations. Although RNA-seq has been demonstrated in many organisms, no study has provided a comprehensive investigation of the bovine transcriptome using RNA-seq. Results In this study, we provide a deep survey of the bovine embryonic transcriptomes, the first application of RNA-seq in cattle. Embryos cultured in vitro were used as models to study early embryonic development in cattle. RNA amplified from limited amounts of starting total RNA were sequenced and mapped to the reference genome to obtain digital gene expression at single base resolution. In particular, gene expression estimates from more than 1.6 million unannotated bases in 1785 novel transcribed units were obtained. We compared the transcriptomes of embryos showing distinct developmental statuses and found genes that showed differential overall expression as well as alternative splicing. Conclusion Our study demonstrates the power of RNA-seq and provides further understanding of bovine preimplantation embryonic development at a fine scale.

  16. PENICILLIN-STREPTOMYCIN IN THE CULTURE MEDIUM DURING IN VITRO MATURATION (IVM OF BOVINE OOCYTES AFFECTS NUCLEAR MATURATION AND SUBSEQUENT EMBRYO DEVELOPMENT

    Directory of Open Access Journals (Sweden)

    SHIRAZI A

    2001-01-01

    Full Text Available Introduction: Standard concentrations of antibiotics in culture media are thought to have no detectable toxic effects on the cultured cells. However, since antibiotics are biologically active substances, the possibility that they interfere to some extent with cellular processes occurring in the cultured cells can not always be totally excluded. This study, therefore, was conducted to assess whether the presence of penicllin-streptomycin (pen-strep during in vitro maturation (IVM of bovine cumulus oocyte complexes (COCs affect nuclear and cytoplasmic maturation and subsequent embryo development. Materials and Methods: Bovine COCs were matured at 39oC in a humidified atmosphere with 5 % CO2 in air for 24 h in: 1- culture medium M 199 supplemented with 10 % FCS (Fetal calf serum, 0.05 IU/ml rhFSH (recombinant human FSH and 100 units penicillin and 100 ?g streptomycin/ ml. 2- culture medium M 199 without FCS and rhFSH in the presence of pen-strep. Cultures without antibiotics served as control. Six series of experiments, each consisted of at least 3 replicates, were performed. Results: In vitro maturation in the presence of pen-strep in culture medium supplemented with FCS and rhFSH significantly (P<0.05 increased the percentage of MII oocytes, however, when the COCs were divided, on the basis of appearance of the cumulus investment, into bright and dark groups, this effect was less obvious in both types of COCs, 76% vs 72% in bright COCs (P= 0.149 or 83% vs 80% in dark COCs (P=0.296 in treated and control groups respectively. The percentage of oocytes with type III of cortical granules (CGs distribution was not affected in the presence of pen-strep. The COCs expansion after IVM was not affected by the presence of antibiotics in culture medium. The subsequent embryo development of IVM/IVF produced ova, which were exposed to pen-strep during IVM, was significantly (P<0.05 decreased with respect to blastocyst formation by day 9. In vitro maturation in

  17. Culture of bovine embryos in polyester mesh sections: the effect of pore size and oxygen tension on in vitro development.

    Science.gov (United States)

    Somfai, T; Inaba, Y; Aikawa, Y; Ohtake, M; Kobayashi, S; Akai, T; Hattori, H; Konishi, K; Imai, K

    2010-12-01

    The purpose of this study was to assess the feasibility of polyester mesh culture for the in vitro production of bovine embryos, as polyester mesh is an alternative way for tracking individual embryos throughout culture using time-lapse cinematography (TLC). Bovine embryos were isolated during in vitro culture using sections of three different polyethylene terephthalate (PET) mesh products. In vitro matured and fertilized bovine oocytes were cultured in the 217 × 217, 230 × 230 or 238 × 238-μm openings of PET mesh sections or in simple micro-drops (control) for 7 days under either 20% or 5% O(2) tensions. No difference in embryo developmental rates was found between the culture groups in terms of cleavage, blastocyst formation and blastocyst expansion irrespective of O(2) tension. In contrast, under 20% O(2) tension, blastocysts that developed in PET mesh with 217 × 217-μm opening had significantly higher numbers of total and trophectoderm (TE) cells than control embryos; however, the numbers and proportions of inner cell mass (ICM) cells did not differ. Under 5% O(2) tension, no difference was found among the culture groups in the numbers of total, ICM and TE cells in embryos. All three PET mesh products investigated in this study were proven to be effective to prevent embryo movement. The results demonstrate that bovine embryos can be cultured in PET mesh sections without negative side-effects and suggest that embryo distance determined by the mesh affects embryo quality at atmospheric oxygen tension. Polyethylene terephthalate mesh with 217 × 217-μm openings was found to be the most suitable for further application in TLC. PMID:19845884

  18. Effect of temporary meiosis block during prematuration of bovine cumulus-oocyte complexes on pregnancy rates in a commercial setting for in vitro embryo production.

    Science.gov (United States)

    Guemra, Samuel; da Silva Santo, Eriko; Zanin, Renato; Monzani, Paulo Sergio; Sovernigo, Tobias Canan; Ohashi, Otávio Mitio; Verde Leal, Cláudia Lima; Adona, Paulo Roberto

    2014-04-15

    Ovum pick up (OPU) associated with in vitro production (IVP) of embryos has been shown as an important tool in cattle breeding to increase the number of descendants from animals of high genetic value. In herds maintained distant from the laboratory, collecting cumulus-oocyte complexes (COCs) and transporting them to the laboratory may take several hours and decrease COCs viability, representing a challenge for commercial settings. In this study, a prematuration culture to induce temporary meiosis block was evaluated in a commercial scale IVP setting as a strategy to transport bovine OPU-derived COCs from Nelore and Brangus donors. Effects on embryo yield and pregnancy rates were assessed. Viable COCs from each donor were destined to one of the experimental groups (control, blocks 1 and 2). Control group COCs were placed in cryotubes with 1 mL TCM199-HEPES. In block groups (1 and 2), COCs were placed in cryotubes with 300 μL TCM 199 + 12 μM butyrolactone I (block medium). All groups were gassed and kept in a thermos bottle for 4 hours at 36 °C. Next, COCs in the control group were transferred to IVM medium and block 1 group to block medium, and cultured for 22 hours and 15 hours, respectively, at 38.5 °C and 5% CO2 in air. Block 2 COCs were kept in the cryotubes and in the thermos bottle for another 15 hours at 36 °C to simulate long-term transport conditions. After meiosis block in prematuration culture, blocks 1 and 2 COCs were matured in vitro for 22 hours as for the control group. After IVM, COCs in all groups were submitted to IVF and IVC, and blastocyst rates were evaluated on day 7. Embryos were transferred and pregnancy rates evaluated at 60 days of gestation. The mean total number of COCs retrieved by OPU did not differ between Nelore and Brangus donors (16.8 and 17.2, respectively, P > 0.05), but Nelore donors produced more viable COCs than Brangus (10.1 and 7.6, respectively, P Brangus cattle, respectively (P > 0.05). Pregnancy rates did

  19. Homologous recombination and non-homologous end-joining repair pathways in bovine embryos with different developmental competence

    International Nuclear Information System (INIS)

    This study investigated the expression of genes controlling homologous recombination (HR), and non-homologous end-joining (NHEJ) DNA-repair pathways in bovine embryos of different developmental potential. It also evaluated whether bovine embryos can respond to DNA double-strand breaks (DSBs) induced with ultraviolet irradiation by regulating expression of genes involved in HR and NHEJ repair pathways. Embryos with high, intermediate or low developmental competence were selected based on the cleavage time after in vitro insemination and were removed from in vitro culture before (36 h), during (72 h) and after (96 h) the expected period of embryonic genome activation. All studied genes were expressed before, during and after the genome activation period regardless the developmental competence of the embryos. Higher mRNA expression of 53BP1 and RAD52 was found before genome activation in embryos with low developmental competence. Expression of 53BP1, RAD51 and KU70 was downregulated at 72 h and upregulated at 168 h post-insemination in response to DSBs induced by ultraviolet irradiation. In conclusion, important genes controlling HR and NHEJ DNA-repair pathways are expressed in bovine embryos, however genes participating in these pathways are only regulated after the period of embryo genome activation in response to ultraviolet-induced DSBs.

  20. Homologous recombination and non-homologous end-joining repair pathways in bovine embryos with different developmental competence

    Energy Technology Data Exchange (ETDEWEB)

    Henrique Barreta, Marcos [Universidade Federal de Santa Catarina, Campus Universitario de Curitibanos, Curitibanos, SC (Brazil); Laboratorio de Biotecnologia e Reproducao Animal-BioRep, Universidade Federal de Santa Maria, Santa Maria, RS (Brazil); Garziera Gasperin, Bernardo; Braga Rissi, Vitor; Cesaro, Matheus Pedrotti de [Laboratorio de Biotecnologia e Reproducao Animal-BioRep, Universidade Federal de Santa Maria, Santa Maria, RS (Brazil); Ferreira, Rogerio [Centro de Educacao Superior do Oeste-Universidade do Estado de Santa Catarina, Chapeco, SC (Brazil); Oliveira, Joao Francisco de; Goncalves, Paulo Bayard Dias [Laboratorio de Biotecnologia e Reproducao Animal-BioRep, Universidade Federal de Santa Maria, Santa Maria, RS (Brazil); Bordignon, Vilceu, E-mail: vilceu.bordignon@mcgill.ca [Department of Animal Science, McGill University, Ste-Anne-De-Bellevue, QC (Canada)

    2012-10-01

    This study investigated the expression of genes controlling homologous recombination (HR), and non-homologous end-joining (NHEJ) DNA-repair pathways in bovine embryos of different developmental potential. It also evaluated whether bovine embryos can respond to DNA double-strand breaks (DSBs) induced with ultraviolet irradiation by regulating expression of genes involved in HR and NHEJ repair pathways. Embryos with high, intermediate or low developmental competence were selected based on the cleavage time after in vitro insemination and were removed from in vitro culture before (36 h), during (72 h) and after (96 h) the expected period of embryonic genome activation. All studied genes were expressed before, during and after the genome activation period regardless the developmental competence of the embryos. Higher mRNA expression of 53BP1 and RAD52 was found before genome activation in embryos with low developmental competence. Expression of 53BP1, RAD51 and KU70 was downregulated at 72 h and upregulated at 168 h post-insemination in response to DSBs induced by ultraviolet irradiation. In conclusion, important genes controlling HR and NHEJ DNA-repair pathways are expressed in bovine embryos, however genes participating in these pathways are only regulated after the period of embryo genome activation in response to ultraviolet-induced DSBs.

  1. GnRH agonist versus GnRH antagonist in in vitro fertilization and embryo transfer (IVF/ET).

    Science.gov (United States)

    Depalo, Raffaella; Jayakrishan, K; Garruti, Gabriella; Totaro, Ilaria; Panzarino, Mariantonietta; Giorgino, Francesco; Selvaggi, Luigi E

    2012-04-13

    Several protocols are actually available for in Vitro Fertilization and Embryo Transfer. The review summarizes the main differences and the clinic characteristics of the protocols in use with GnRH agonists and GnRH antagonists by emphasizing the major outcomes and hormonal changes associated with each protocol. The majority of randomized clinical trials clearly shows that in "in Vitro" Fertilization and Embryo Transfer, the combination of exogenous Gonadotropin plus a Gonadotropin Releasing Hormone (GnRH) agonist, which is able to suppress pituitary FSH and LH secretion, is associated with increased pregnancy rate as compared with the use of gonadotropins without a GnRH agonist. Protocols with GnRH antagonists are effective in preventing a premature rise of LH and induce a shorter and more cost-effective ovarian stimulation compared to the long agonist protocol. However, a different synchronization of follicular recruitment and growth occurs with GnRH agonists than with GnRH antagonists. Future developments have to be focused on timing of the administration of GnRH antagonists, by giving a great attention to new strategies of stimulation in patients in which radio-chemotherapy cycles are needed.

  2. GnRH agonist versus GnRH antagonist in in vitro fertilization and embryo transfer (IVF/ET

    Directory of Open Access Journals (Sweden)

    Depalo Raffaella

    2012-04-01

    Full Text Available Abstract Several protocols are actually available for in Vitro Fertilization and Embryo Transfer. The review summarizes the main differences and the clinic characteristics of the protocols in use with GnRH agonists and GnRH antagonists by emphasizing the major outcomes and hormonal changes associated with each protocol. The majority of randomized clinical trials clearly shows that in “in Vitro” Fertilization and Embryo Transfer, the combination of exogenous Gonadotropin plus a Gonadotropin Releasing Hormone (GnRH agonist, which is able to suppress pituitary FSH and LH secretion, is associated with increased pregnancy rate as compared with the use of gonadotropins without a GnRH agonist. Protocols with GnRH antagonists are effective in preventing a premature rise of LH and induce a shorter and more cost-effective ovarian stimulation compared to the long agonist protocol. However, a different synchronization of follicular recruitment and growth occurs with GnRH agonists than with GnRH antagonists. Future developments have to be focused on timing of the administration of GnRH antagonists, by giving a great attention to new strategies of stimulation in patients in which radio-chemotherapy cycles are needed.

  3. HCG注射日和胚胎移植日血清雌二醇及孕酮与IVF/ICSI结局的关系%Oestradiol and progesterone concentration on the day of human chononic gonadotrophin administration and embryo implantation and their relationship with IVF/ICSI outcome

    Institute of Scientific and Technical Information of China (English)

    王芬; 曹云霞; 魏兆莲; 周平

    2009-01-01

    目的 探讨hCG注射日血清雌二醇(hCG-E2)和胚胎移植日血清雌二醇(ET-E2)、孕酮(P)及他们之间的比值与IVF/ICSI结局的关系.方法 对182个采用促性腺激素激动剂/促卵泡生成素或(和)人绝经期促性腺激素/绒毛膜促性腺激素(GnRH-a/FSH/HMG/hCG)长方案超促排卵的体外受精(IVF)和单精子胞浆内注射(ICSI)的周期,在hCG注射日和胚胎移植日采用化学发光免疫法测定E2和P.结果 (1)hCG-E2波动在8000~20000pmol/l时,妊娠率最高(46.8%),与E220000pmol/l两组相比均有统计学差异(P<0.05).(2)移植日E2下降的水平越大.即hCG-E2/ET-E2越大,妊娠率越低.(3)ET日P/E2越高临床妊娠率越低,但排除E2的影响后,比值无意义.结论 hCG注射日和ET日E2及其比值对IVF/ICSI的结局可能有预测价值.%Objective: To study plasma oestradiol, progesterone and their ratio on the day of human chorionic gonadotrophin (HCG) and the day of embryo implantation, and the relationship with IVF/ICSI outcome. Methods: 182 women underwent 182 IVF/ ICSI cycles were prospectively analysed. They were employed controll oocytes hyperstimulation (COH) with a long protocol of gonadotrophin-releasing hormane agonist (GnRH-a) /follicle-stimulating hormone (FSH) or (and) human menopausal gonadotrophin (HCG) /human chorionic gonadotrophin (HCG). Plasma oestradiol and progesterone on the day of human chorionic gonadotrophin (HCG) and the day of embryo implantation were measured by chemiluminescence immunoassayed. Results: (1) When plasma oestradiol level on the day of human chorionic gonadotrophin fluctuated in 8000 ~ 20000pmol/1, the pregnancy rate was the highest (46. 8%). (2) The falling oestradiol on the day of embryo implantation was associated with poor outcome. (3) Regardless of the difference of estradiol , the ratio of progesterone to estradiol on the day of embryos implantion was negative relation with clinical preganancy rate. Conclution: Plasma oestradiol on the day of

  4. The effect of IVM and IVC media on in vitro development of bovine embryos

    Directory of Open Access Journals (Sweden)

    E.T Mergawati

    2000-12-01

    Full Text Available The purpose of this study was to examine the effect of medium combination of IVM and IVC on the in vitro development of bovine embryos. The study involved 4 groups in a 2 (IVM medium x 2 (IVC medium factorial in a randomized block design. Each group was replicated for 5 times. The treatments were as follows: TCM-199/CR1aa (T1; TCM-199/SOF (T2; B- 199/CR1aa (T3 and B-199/SOF (T4. Oocytes were aspirated from ovaries collected at local abattoirs using aspiration medium of PBS supplemented with 3% FCS and 0.1% Penicillin and Streptomycin. The oocytes were matured in medium of TCM-199 or B-199 supplemented with 10% FCS, hormones: 10μg/ml FSH+ 10μg/ml hCG+ 1μg/ml Estradiol. Maturation was maintained at 37oC for 22 hours in 5% CO2 incubator with high humidity. A method of BRACKETT & Oliphant (BO was used to fertilize the matured oocytes. The fertilization was incubated for 7 hours in the 5% CO2 incubator. Two culture media of CR1aa or SOF/AA/BSA were used to develop the fertilized oocytes undergo to morula and blastocyst embryos. The findings showed that the proportion of oocytes cleaved and formation of blastocysts were affected significantly by a combination of IVM and IVC media (P<0.05. A combination of B-199/SOF (T4 resulted in a higher blastocyst rate (32% than others (T3= 29%; T2=T1= 23%. This study suggests that either SOF/AA/BSA or CR1aa has similar competence in development of bovine embryos in vitro.

  5. Tumor necrosis factor alpha inhibits in vitro bovine embryo development through a prostaglandin mediated mechanism

    Directory of Open Access Journals (Sweden)

    Jackson Lauren R

    2012-03-01

    Full Text Available Abstract Mastitis or other infectious diseases have been related to reduced fertility in cattle. Inflammatory cytokines such as tumor necrosis factor α (TNFα are released in response to infection and may have negative effects on embryo development. In the current study the effect of exposure to TNFα on the development of in vitro fertilized bovine embryos was examined. Indomethacin, a prostaglandin synthesis inhibitor, was used to determine if blockade of prostaglandin synthesis would alter the effects of TNFα. Ovaries were obtained from a local abattoir and immature COC were isolated from 2-10 mm follicles, in vitro matured and fertilized. After fertilization, groups of presumptive zygotes were randomly placed into either control development medium, medium containing 25 ng/mL TNFα or medium containing 25 ng/mL TNFα plus 1 μg/mL indomethacin. The proportion of blastocysts formed was assessed at day 7 of culture. Fewer embryos exposed to TNFα alone reached the blastocyst stage (17.5 ± 2.4%, P

  6. In Vitro Developmental Potential of Cloned Embryos Derived from Bovine Somatic Cells and Rabbits Oocyte

    Institute of Scientific and Technical Information of China (English)

    LIU Ya; LI Bin; ZHAO Huan; CHENG Li-zi; ZHANG Xiao-rong; CHEN Da-yuan; ZHANG Yun-hai; ZHANG Zhi-guo; JING Ren-tao; WANG Cun-li; ZHANG Mei-lin; LI Dong-wei

    2003-01-01

    180 reconstituted embryos were produced by nuclear transplantation using bovine ear fibroblasts at G0 or non-G0 stage as donor nuclei and oocytes collected from superovulated multiparous or young rabbits as recipients. After cultivation in two kinds of medium M199+ 10%FBS or RD+ 10%FBS, 112 of them developed to 2-cell stage (62.2%) and 26 to morula stage (14.4%) and 20 of them eventually developed to blastocyst stage (11. 1% ). There is no significant difference for the cleavage rates in two groups of reconstituted embryos derived from G0-stage and non-G0 stage donor cells respectively. However, G0-stage donor cells could result in higher rate of 8-cell - 16-cell stage embryos significantly (P<0.05), as well as higher rate of blastocysts (P<0.01). It seems that using two different culture systems had no significant effects on the cleavage rate, morula rate or blastocyst rate (P>0.05).

  7. Monitoring in-vitro bovine embryo development during the first days after fertilization (Conference Presentation)

    Science.gov (United States)

    Kandel, Mikhail E.; Rubessa, Marcello; Fernandes, Daniel; Nguyen, Tan H.; Wheeler, Matthew B.; Popescu, Gabriel

    2016-03-01

    Conventional label-based contrast enhancement techniques (e.g., fluorescence) frequently modify the genetic makeup of tagged cells, making them poor candidates for use in in-vitro fertilization applications. Instead, we choose a label-free form of contrast, based on interferometric imaging, sensitive to optical path length differences. Compared to, single HeLa cells, typical mammalian ova and embryos are more than an order of magnitude thicker. As a result, regions of large phase variation lead to phase wrapping and an overall reduction in signal intensity occurs due to multiple scattering. These effects manifest themselves in low-spatial frequencies (blurs), with the desired details buried in the background. We present a phase shifting interferometer that yields the derivative of the phase, a quantity whose value is particularly sensitive to local variations and fine details. We demonstrate that our new real-time imaging platform is valuable in measuring the multiday development of bovine embryos. Reconstructing the derivative of the image phase and amplitude, we characterize the motion of previously low-contrast structures, which are relevant for embryo viability tests.

  8. Cloned embryos from semen. Part 2: Intergeneric nuclear transfer of semen-derived eland (Taurotragus oryx) epithelial cells into bovine oocytes

    Science.gov (United States)

    Nel-Themaat, L.; Gomez, M.C.; Pope, C.E.; Lopez, M.; Wirtu, G.; Jenkins, J.A.; Cole, A.; Dresser, B.L.; Bondioli, K.R.; Godke, R.A.

    2008-01-01

    The production of cloned offspring by nuclear transfer (NT) of semen-derived somatic cells holds considerable potential for the incorporation of novel genes into endangered species populations. Because oocytes from endangered species are scarce, domestic species oocytes are often used as cytoplasts for interspecies NT. In the present study, epithelial cells isolated from eland semen were used for intergeneric transfer (IgNT) into enucleated bovine oocytes and compared with bovine NT embryos. Cleavage rates of bovine NT and eland IgNT embryos were similar (80 vs. 83%, respectively; p > 0.05); however, development to the morula and blastocyst stage was higher for bovine NT embryos (38 and 21%, respectively; p bovine NT or eland IgNT cybrids before activation, but in 75 and 70% of bovine NT and eland igNT embryos, respectively, cell-cycle resumption was observed at 16 h postactivation (hpa). For eland IgNT embryos, 13% had ???8 cells at 84 hpa, while 32% of the bovine NT embryos had ???8 cells at the same interval. However, 100 and 66% of bovine NT and eland IgNT embryos, respectively, that had ???8 cells synthesized DNA. From these results we concluded that (1) semen-derived epithelial cell nuclei can interact and be transcriptionally controlled by bovine cytoplast, (2) the first cell-cycle occurred in IgNT embryos, (3) a high frequency of developmental arrest occurs before the eight-cell stage in IgNT embryos, and (4) IgNT embryos that progress through the early cleavage stage arrest can (a) synthesize DNA, (b) progress through subsequent cell cycles, and (c) may have the potential to develop further. ?? 2008 Mary Ann Liebert, Inc.

  9. Cellular and molecular deviations in bovine in vitro-produced embryos are related to the large offspring syndrome

    NARCIS (Netherlands)

    Lazzari, G.; Wrenzycki, C.; Herrmann, D.; Duchi, R.; Kruip, T.; Niemann, H.; Galli, C.

    2002-01-01

    The large offspring syndrome (LOS) is observed in bovine and ovine offspring following transfer of in vitro-produced (IVP) or cloned embryos and is characterized by a multitude of pathologic changes, of which extended gestation length and increased birthweight are predominant features. In the presen

  10. Effect of exogenous glutathione in medium fertilization to improve blastocyst rates of bovine embryos

    Directory of Open Access Journals (Sweden)

    E Triwulaninngsih

    2002-06-01

    Full Text Available Glutathione (C10H17N3O6S is a tripeptide (γ-Glu-Cys-Gly widespread in living organism. Glutathione (GSH at the 5 mM concentration increased the motility and fertility of frozen-thawed sperm. Intracellulair glutathione improved the cleavage rate and embryo development to the blastocyst rate. Research on in vitro embryos production through the implementation of GSH during IVF (in vitro fertilization on embryo development has been conducted at the Laboratorium Reproductive of Physiology, Research Institute for Animal Production. Ovaries of beef cattle as source of oocytes were collected from the slaughterhouse in a thermo flask with 350C PBS as medium and transported to the laboratory. The oocytes were fertilized in vitro with selected motile sperm using Percoll gradient (90 and 45%. Ten COCs (cumulus oocytes complexes were transfered to 44 μl of fertilization medium (mTALP was performed with either 0; 0.25; 0.50; 0.75 and 1.00 mM of glutathione as treatment A, B, C, D and E respectively, and inseminated with 2 μl of capacitated sperm and added 2 μl of heparin and 2 μl of PHE (consisting of 20 μM penicillamine, 10 μM hypotaurine, 1 μM epinephrine. Incubation between sperm and oocytes in the 5% CO2 incubator and RH 90% in fertilization media (TALP for 20 hours. All zygotes were cultured in modification of CR1aa medium up to blastocyst and were fed serum 5 μl/50μl medium on day 6. Results of this experiment showed that the effect of concentration of glutathione (0, 0.25; 0.50; 0.75 and 1.00 mM on fertilization medium to the percentage of cleavage rates were 69.35 + 29.40; 79.07 + 13.17; 67.88 + 10.65; 98.10 + 3.30 and 82.62 + 24.19% not significant different (P>0.05 among treatments A, B, C, D dan E respectively.The precentages of morula and blastocyst for treatment D were 50.45 + 42.64% and 31.28 + 24.28%; while 36.55 + 24.13% and 17.74 + 17.86% for treatment E respectively.

  11. Risk assessment of transmission of bovine viral diarrhea virus (BVDV) in abattoir-derived in vitro produced embryos.

    Science.gov (United States)

    Perry, G H

    2007-07-01

    Bovine virus diarrhea virus (BVDV) is a pathogen of the bovine reproductive system causing reduced conception rates, abortions and persistently infected calves. Most if not all strains of BVDV are transmissible by natural mating and AI. For international trade, it is recommended that in vitro fertilized embryos be washed according to the IETS Manual. However, BVDV may not be entirely washed out, resulting in possible transmission risks to recipients. Donor cows, donor bulls and biological agents are all possible sources of contamination. The process for producing in vitro produced (IVP) embryos is complex and non-standard, and some procedures can contribute to spread of BVDV to uninfected embryos. The structure of the zone pellucida (ZP) of IVP embryos permits adherence of BVDV to the ZP. To estimate the risk of producing infected recipients and persistently infected calves from abattoir-derived IVP embryos, a quantitative risk assessment model using Microsoft Excel and Palisade @Risk was developed. Assumptions simplified some of the complexities of the IVP process. Uncertainties due to incomplete or variable data were addressed by incorporating probability distributions in the model. Model variables included: disease prevalence; the number of donor cows slaughtered for ovaries; the number of oocytes collected, selected and cultured; the BVDV status of ovaries, semen, biological compounds and its behavior in the IVP embryo process. The model used the Monte Carlo method to simulate the IVP process. When co-culture cells derived from donor cows of unknown health status were used for in vitro culture (IVC), the probability of a recipient cow at risk of infection to BVDV per oocyte selected for IVP processing averaged 0.0006. However, when co-culture free from BVDV was used, the probability was 1.2 x 10(-5). Thus, for safe international trade in bovine IVP embryos (i.e. negligible risks of transmission of BVDV), co-culture cells, if used during IVC for producing IVP

  12. RNA-Seq analysis uncovers transcriptomic variations between morphologically similar in vivo- and in vitro-derived bovine blastocysts

    Directory of Open Access Journals (Sweden)

    Driver Ashley M

    2012-03-01

    Full Text Available Abstract Background A valuable tool for both research and industry, in vitro fertilization (IVF has applications range from gamete selection and preservation of traits to cloning. Although IVF has achieved worldwide use, with approximately 339,685 bovine embryos transferred in 2010 alone, there are still continuing difficulties with efficiency. It is rare to have more than 40% of fertilized in vitro cattle oocytes reach blastocyst stage by day 8 of culture, and pregnancy rates are reported as less than 45% for in vitro produced embryos. To investigate potential influences in-vitro fertilization (IVF has on embryonic development, this study compares in vivo- and in vitro-derived bovine blastocysts at a similar stage and quality grade (expanded, excellent quality to determine the degree of transcriptomic variation beyond morphology using RNA-Seq. Results A total of 26,906,451 and 38,184,547 fragments were sequenced for in vitro and in vivo embryo pools, respectively. We detected expression for a total of 17,634 genes, with 793 genes showing differential expression between the two embryo populations with false discovery rate (FDR Conclusions Thus, our results support that IVF may influence at the transcriptomic level and that morphology is limited in full characterization of bovine preimplantation embryos.

  13. Expression of the gap junction gene connexin43 (Cx43) in preimplantation bovine embryos derived in vitro or in vivo.

    Science.gov (United States)

    Wrenzycki, C; Herrmann, D; Carnwath, J W; Niemann, H

    1996-09-01

    In this study we have examined the presence of mRNA encoding connexin 43 (Cx43) in bovine embryos derived in vivo and in vitro. Cumulus-oocyte complexes, immature and matured oocytes liberated from cumulus cells, zygotes, 2-4-cell and 8-16-cell embryos, morulae, blastocysts and hatched blastocysts were produced in vitro from ovaries obtained from an abattoir using TCM 199 supplemented with hormones and 10% oestrous cow serum for maturation. Cumulus-oocyte complexes matured for 24 h were exposed to bull spermatozoa for 19 h and then cultured in TCM 199 supplemented with 10% oestrous cow serum to the desired developmental stage. Morulae and blastocysts derived in vivo were collected from superovulated donor cows. Total RNA was extracted from pools of 60-200 bovine oocytes or embryos using a modified phenol-chloroform extraction method and analysed by reverse transcriptase polymerase chain reaction. Before reverse transcription, aliquots of DNase-digested embryonic RNA were tested by polymerase chain reaction using bovine-specific primers to control for residual genomic DNA contamination. DNA-free, total RNA was reverse transcribed after preincubation with the Cx43 specific 3'primer. The resultant cDNA was amplified by polymerase chain reaction using Cx43 specific primers that define a 516 bp fragment of Cx43. The reverse transcriptase polymerase chain reaction product was verified by restriction enzyme analysis with Alu I and sequencing. Assays were repeated at least twice for each developmental stage and provided identical results between replicates. Cx43 transcripts were detected in bovine morulae and blastocysts grown in vivo. In contrast, whereas the early in vitro stages from cumulus-oocyte complexes to morulae expressed Cx43, blastocysts and hatched blastocysts did not have detectable concentrations of mRNA from this gene. Restriction enzyme cutting revealed three fragments of the predicted size (139, 177, 200 bp). The amplified product showed 100% identity

  14. Co-expression network analysis to identify pluripotency biomarkers in bovine and porcine embryos

    DEFF Research Database (Denmark)

    Mazzoni, Gianluca; Freude, Karla Kristine; Hall, Vanessa Jane;

    stable culture of pluripotent cells in pig and cattle. Methods After quality control reads were pre-processed and mapped with STAR aligner. Post-mapping quality was checked with Qualimap. Finally the expression levels were estimated using HTSeq. Gene co-expression will be analyzed using a weighted...... network based method to identify highly co-expressed genes (module) and hub genes. Modules with a potential role in pluripotency will be identified with enrichment procedure and regulator genes identified with LemonTree algorithm. Differential wiring of modules among species will be evaluated. Expected...... results We expect to find out candidate pluripotency factors in porcine and bovine embryo. Acknowledgements We thank for the financial support from the EU project PluriSys, HEALTH-2007-B-223485....

  15. A randomized study comparing IVF in the unstimulated cycle with IVF-following clomiphene citrate

    DEFF Research Database (Denmark)

    Ingerslev, Hans Jakob; Højgaard, A.; Hindkjær, Johnny Juhl;

    2001-01-01

    The efficiency of IVF in unstimulated cycles was compared with that following ovarian stimulation with clomiphene citrate in a simple protocol with ultrasound monitoring only. A total of 132 couples with no previous IVF attempts, selected by female age ... protocol, but not IVF in unstimulated cycles, seems compatible with the concept of `friendly IVF', yielding a fair pregnancy rate both per cycle started and per embryo transfer in selected patients. The results do not substantiate any important negative anti-oestrogenic effects of clomiphene....

  16. Effect of growth factors on oocyte maturation and allocations of inner cell mass and trophectoderm cells of cloned bovine embryos.

    Science.gov (United States)

    Arat, Sezen; Caputcu, Arzu Tas; Cevik, Mesut; Akkoc, Tolga; Cetinkaya, Gaye; Bagis, Haydar

    2016-08-01

    This study was conducted to determine the additive effects of exogenous growth factors during in vitro oocyte maturation (IVM) and the sequential culture of nuclear transfer (NT) embryos. Oocyte maturation and culture of reconstructed embryos derived from bovine granulosa cells were performed in culture medium supplemented with either epidermal growth factor (EGF) alone or a combination of EGF with insulin-like growth factor-I (IGF-I). The maturation rates of oocytes matured in the presence of EGF or the EGF + IGF-I combination were significantly higher than those of oocytes matured in the presence of only fetal calf serum (FCS) (P 0.05). IGF-I alone or in combination with EGF in sequential embryo culture medium significantly increased the ratio of inner cell mass (ICM) to total blastocyst cells (P media of cloned bovine embryos increased the ICM without changing the total cell number. These unknown and uncontrolled effects of growth factors can alter the allocation of ICM and trophectoderm cells (TE) in NT embryos. A decrease in TE cell numbers could be a reason for developmental abnormalities in embryos in the cloning system. PMID:26444069

  17. Establishment of conditions for ovum pick up and IVM of jennies oocytes toward the setting up of efficient IVF and in vitro embryos culture procedures in donkey (Equus asinus).

    Science.gov (United States)

    Goudet, Ghylène; Douet, Cécile; Kaabouba-Escurier, Aurore; Couty, Isabelle; Moros-Nicolás, Carla; Barrière, Philippe; Blard, Thierry; Reigner, Fabrice; Deleuze, Stefan; Magistrini, Michèle

    2016-07-15

    Most wild and domestic donkey breeds are currently endangered or threatened. Their preservation includes the creation of a Genome Resource Bank. Embryos cryopreservation allows the preservation of genetics from both male and female and is the fastest method to restore a breed. Because embryo production in vivo is limited in equids, our objective was to establish conditions for in vitro production of embryos in donkey using ovum pick up (OPU), IVM, IVF, and in vitro culture of zygotes. Donkey cumulus-oocyte complexes (COCs) were collected by transvaginal ultrasound-guided aspirations OPU in adult cyclic jennies and in vitro matured in tissue culture medium 199 supplemented with fetal calf serum and epidermal growth factor for 24, 30, 34, or 38 hours. They were preincubated with oviductal fluid for 30 minutes, coincubated with frozen-thawed donkey semen treated with procaine for 18 hours, and cultured for 30 hours in Dulbecco's Modified Eagle Medium-F12 supplemented with NaHCO3, fetal calf serum, and gentamycin. From the five OPU sessions, we collected 92 COCs in 193 follicles (48%) with an average of 4.2 COCs per jenny. All COCs were expanded after more than 24-hour IVM. At collection, jennies oocytes contained a germinal vesicle. Metaphase 1 oocytes were observed after 30-hour IVM and 44% were in metaphase 2 after 34-hour IVM. In our conditions, IVM of donkey oocytes was slower than IVM of equine oocytes and optimal duration for donkey oocytes IVM may be 34 hours. Only 15% of jennies oocytes contained two pronuclei after coincubation with donkey spermatozoa and none of them developed further after 48 hours post-IVF. Moreover, some parthenogenetic activation occurred. Thus, the treatment of donkey sperm with procaine may not be efficient for IVF. In conclusion, we established for the first time conditions for OPU in jennies with high recovery rates. We reported that IVM of jennies oocytes can produce 44% of metaphase 2 oocytes after 34 hours in culture

  18. 牛胚胎的体外生产技术%In vitro Bovine Embryo Production

    Institute of Scientific and Technical Information of China (English)

    马云; 窦忠英

    2001-01-01

    应用体外受精技术,可以有效地利用淘汰的高产奶牛和屠宰的青年母牛卵巢中的未成熟卵,大量、廉价地实验室生产胚胎。这对加速我国高产奶牛群和良种肉牛群的繁殖,是一项行之有效的胚胎工程技术。借鉴国外实验室生产牛胚胎的经验和根据我们实验室的实践,本文比较详细地叙述了体外受精牛胚胎(试管牛)的生产过程,包括卵母细胞的获得、培养、精子获能、体外受精以及受精后的胚胎培养、超低温冷冻保存与解冻后的胚胎移植等。%Using the immature ovum from eliminated high production dairy cow and slaughtered young cow, a number of embryos can be produced by in vitro insemination, it is an effective method to speed reproducing of high production dairy cow and improved beef breeds. According to abroad experience and our practices, the procedure of bovine embryo production in vitro, including oocyte collection, in vitro cultivation, sperm capacitation, in vitro insemination and embryo cultivation, freezing, thaw and transfer were reviewed in this paper.

  19. Psychological IVF

    DEFF Research Database (Denmark)

    Adrian, Stine Willum

    2015-01-01

    During ethnographic fieldwork at a fertility clinic in Denmark, I became intrigued by emotions. In particular, I found an incidence labelled ‘psychological IVF’ theoretically provocative as it challenged my views on materializations, which I was preparing to study. This paper centres on the story...... of psychological IVF, and I use this narrative to consider emotions and materialization methodologically. I also ask how emotions at fertility clinics can be conceptualized to enable analysis of their materialization, change, and effects. In order to do so, I develop the term ‘emotional choreography......’. This theoretical work has three aims. First, it seeks to illustrate how the story of psychological IVF offers a rich range of materializations of emotions. Secondly, this work proposes a feminist materialist conceptualization of emotions that is both non-representational and posthuman. This conceptualization draws...

  20. Holistic differential analysis of embryo-induced alterations in the proteome of bovine endometrium in the preattachment period.

    Science.gov (United States)

    Berendt, Frank J; Fröhlich, Thomas; Schmidt, Susanne E M; Reichenbach, Horst-Dieter; Wolf, Eckhard; Arnold, Georg J

    2005-07-01

    During the peri-implantation period, molecular signaling between embryo and endometrium (layer of tissue lining the uterus lumen) is supposed to be crucial for the maintenance of pregnancy. To investigate embryo-induced alterations in the proteome of bovine endometrium in the preattachment period (day 18), we used monozygotic cattle twins (generated by embryo splitting) as a model eliminating genetic variability as a source for proteome differences. One of the twins was pregnant after the transfer of two in vitro produced blastocysts, while the corresponding twin received a sham-transfer and served as a nonpregnant control. The two-dimensional fluorescence difference gel electrophoresis (2-D DIGE) analysis of the endometrium samples of three twin pairs (pregnant/nonpregnant) revealed four proteins with significantly higher abundance (p embryo-maternal interactions.

  1. Clinical outcomes of prolonged gonadotropin-releasing hormone agonist(GnRH-a) therapy used in patients with good ovarian reserves and previous in vitro fertilization or intracytoplasmic sperm injection embryo transfer(IVF/ICSI-ET) failure cycle%超长方案在卵巢储备良好前次IVF/ICSI-ET失败患者中的应用分析

    Institute of Scientific and Technical Information of China (English)

    聂玲; 伍琼芳; 张寅; 陈晶晶

    2011-01-01

    Objective : To evaluate the clinical outcomes of prolonged gonadotropm-releasing hormone agonist therapy( GnRH-a) used in patients with good ovanan reserves and previous in vitro fertilization or intracytoplasmic sperm injection embryo transfer( IVF/ICSI-ET) failure cycle. Methods : Retrospective analyze a total of 246 patients with previous IVF-ET failure cycle and good ovarian reserve who underwent the secondary IVF-ET cycles. Patients were divided into two groups : the study group included 52 patients applied with prolonged protocol;the control group included 194 patients applied with long protocol. Results : The numher of oocytes retrieved and the duration of gonadotropin ( Gn) were significantly higher in the study group(P<0. 01). But there were no significant differences in the dose of Gn,total numher of good quality embryos, mean endometrium thickness on transfer day ,mean serum P level on human chorionic gonadotrophin ( HCG) day and severe ovarian hyperstimulation syndrome ( OHSS) rate between the two groups (P> 0. 05). The positive HCG rate and clinical pregnancy rate( CPR) increased obviously in the study group, respectively ( 79. l% vs 61. 1% , P =0. 03 ;69. 8% vs 54. 9% , P=O. 048 ) . And there was an mcreasing tendency in implantation rate ( 39. 5% vs 31. 5% , P = 0. 18 ) . Conclusion : Using prolonged gonadotropin-releasing hormone agonist therapy in patients with good ovarian reserves and previous IVF-ET failure cycle may make the endometrium receptivity better than using the long protocol, and improve obviously the clinical pregnancy outcomes. So the prolonged protocol is recommendable.%目的:评估超长降调节方案在卵巢储备良好前次IVF/ICSI-ET失败患者中的应用效果.方法:回顾分析246例卵巢储备功能良好前次IVF/ICSI-ET常规长方案失败再次行助孕治疗的患者,其中超长方案治疗52例为研究组,常规长方案治疗194例为对照组.结果:研究组Gn时间及

  2. Indication of whole embryo freezing for PCOS patients undergoing long protocol in IVF/ICSI-ET%多囊卵巢综合征患者长方案选择及全胚冷冻指征探讨

    Institute of Scientific and Technical Information of China (English)

    唐蓉; 丁玲玲; 盛燕; 高芹; 党玉洁; 郭情情; 陈子江

    2012-01-01

    Objective: To explore the indication of whole embryos freezing for patients with polycystic ovary syndrome ( PCOS ) in in-vitro fertilization/intracytoplasmic sperm injection-embryo transfer OVF/ICSI-ET). Methods: The data of 1,773 long protocol cycles for PCOS patients who underwent IVF/ICSI-ET from Jan. 2009 to Dec. 2011 were analyzed retrospectively in Hospital for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University. The patients were divided into two groups, the whole embryos freezing group (343 cycles) and the fresh embryos group (1,430 cycles). Besides, they were divided into four subgroups according to the occurrence of moderate or severe ovarian hyper-stimulation syndrome (OHSS). The women in Subgroup A suffered from OHSS after whole embryos freezing procedure (67 cycles), while women in Subgroup B did not (276 cycles). Furthermore, the patients in Subgroup C suffered OHSS after fresh embryos transfer (22 cycles) while patients in Subgroup D did not (1408 cycles). Main outcome measures such as the duration of gonadotrophin administration, the number of oocytes retrieved, the estradiol (E2 ) levels after hCG injection, the number of frozen embryos, clinical pregnancy rate and OHSS incidence were compared among these groups mentioned above. Results: (1). The OHSS incidence of fresh embryos group (1. 5%) was significantly lower than the whole embryos freezing group (19. 5%)(P<0. 01). The clinical pregnancy rate (57%) in fresh embryo transfer group was lower than that in the frozen embryos transfer group (Pembryos transfer group were significantly higher than those in the whole embryos freezing group (P

  3. 手术剥除子宫内膜异位囊肿对IVF-ET结局的影响%Effect of Surgical Treatment for Endometrioma on In Vitro Fertilization-Embryo Transfer Outcomes

    Institute of Scientific and Technical Information of China (English)

    姬萌霞; 赵晓明; 孙赟; 洪燕; 高敏芝; 郑中

    2013-01-01

    Objective: To investigate the effect of endometrioma and stripping endometrioma on IVF-ET outcomes. Methods: A total of 305 cases receiving the first IVF cycle in our center were included in this retrospective study. They were divided into three groups, endometrioma-operated group, endometrioma group, tubal infertility group as the control. All these patients were divided into 2 subgroups according to different ovarian hyper-stimulation protocols (long protocol and short protocol). The outcomes of IVF were compared among the three groups. Results: In both protocols, the average number of retrieved oocytes in endometrioma-operated group was lower than that of endometrioma group and tubal infertility group (P ≤0.001). The endometrioma-operated group was also consuming significantly higher doses of FSH than that of the control (P<0.001). Meanwhile, the pregnancy rate of endometrioma-operated group was lower than that of endometrioma group, the pregnancy rate of both the two groups was lower than that of the control (P<0.05). Compared with the control, the endometrioma group consumed higher FSH dosage (P<0.001), but the average number of retrieved oocytes and total number of embryos were comparable between the two groups. The three groups had no significant difference in the rate of good-quality embryos. In addition, in the endometrioma-operated group, the average number of retrieved oocytes from the operated ovaries was lower than that from the contralateral ovaries (P<0.01), 24% of these patients did not retrieve eggs. Conclusion: Patients with endometrioma have poorer IVF outcomes, and surgical treatment for endometrioma cannot improve the IVF outcome, but to aggravate the damage to ovarian response.%目的:探讨子宫内膜异位囊肿手术剥除与否对IVF-ET结局的影响.方法:回顾性分析接受IVF-ET第1周期治疗的305例患者,分成子宫内膜异位囊肿手术组(A组)、子宫内膜异位囊肿未手术组(B组)和管性不孕对照组(C组),

  4. Post-hatching development of the porcine and bovine embryo - defining criteria for expected development in vivo and in vitro

    DEFF Research Database (Denmark)

    Vejlsted, Morten; Du, Yutao; Vajta, Gabor;

    2006-01-01

    without the need for transfer to recipient animals. Such a system would require (1) definition of milestones of expected post-hatching embryonic development in vivo; and (2) development of adequate culture systems. We propose a stereomicroscopical staging system for post-hatching embryos defining......) Somite stage(s) where paraxial mesoderm gradually condensates to form somites. Post-hatching development of bovine embryos in vitro is compromised and although hatching occurs and elongation can be physically provoked by culture in agarose tunnels, the embryonic disk characterizing the pre-streak stage 1...

  5. Paternal breed effects on expression of IGF-II, BAK1 and BCL2-L1 in bovine preimplantation embryos

    DEFF Research Database (Denmark)

    Valleh, Mehdi Vafaye; Tahmoorespur, Mojtaba; Joupari, Morteza Daliri;

    2015-01-01

    Summary The effects of the paternal breed on early embryo and later pre- and postnatal development are well documented. Several recent studies have suggested that such paternal effects may be mediated by the paternally induced epigenetic modifications during early embryogenesis. The objective...... of this study was to investigate the effects of the paternal breed on the early embryonic development and relative expression of the maternally imprinted gene, IGF-II, and the apoptosis-related genes BAK1 and BCL2-L1 in in vitro produced (IVP) bovine embryos derived from two unrelated paternal breeds (Holstein...

  6. Molecular Characterization of the First Bovine Herpesvirus 4 (BoHV-4 Strains Isolated from In Vitro Bovine Embryos production in Argentina.

    Directory of Open Access Journals (Sweden)

    Erika González Altamiranda

    Full Text Available Bovine herpesvirus 4 (BoHV-4 is increasingly considered as responsible for various problems of the reproductive tract. The virus infects mainly blood mononuclear cells and displays specific tropism for vascular endothelia, reproductive and fetal tissues. Epidemiological studies suggest its impact on reproductive performance, and its presence in various sites in the reproductive tract highlights its potential transmission in transfer-stage embryos. This work describes the biological and genetic characterization of BoHV-4 strains isolated from an in vitro bovine embryo production system. BoHV-4 strains were isolated in 2011 and 2013 from granulosa cells and bovine oocytes from ovary batches collected at a local abattoir, used as "starting material" for in vitro production of bovine embryos. Compatible BoHV-4-CPE was observed in the co-culture of granulosa cells and oocytes with MDBK cells. The identity of the isolates was confirmed by PCR assays targeting three ORFs of the viral genome. The phylogenetic analyses of the strains suggest that they were evolutionary unlinked. Therefore it is possible that BoHV-4 ovary infections occurred regularly along the evolution of the virus, at least in Argentina, which can have implications in the systems of in vitro embryo production. Thus, although BoHV-4 does not appear to be a frequent risk factor for in vitro embryo production, data are still limited. This study reveals the potential of BoHV-4 transmission via embryo transfer. Moreover, the high variability among the BoHV-4 strains isolated from aborted cows in Argentina highlights the importance of further research on the role of this virus as an agent with the potential to cause reproductive disease in cattle. The genetic characterization of the isolated strains provides data to better understand the pathogenesis of BoHV-4 infections. Furthermore, it will lead to fundamental insights into the molecular aspects of the virus and the means by which these

  7. Current ethical issues in IVF.

    Science.gov (United States)

    Grobstein, C; Flower, M

    1985-12-01

    This article has briefly reviewed the range of public policy issues and ethical questions raised with respect to IVF. It then discussed selected issues that are now under policy debate and decision. Given the wide acceptance of IVF as a medical procedure for married couples, what variants might also be ethically defensible? IVF for unmarried couples appears defensible under specific conditions that are equally applicable to married couples. Involvement of third parties (gamete donation and gestational surrogacy) is more complex and needs case by case examination. Sperm donation appears to generate little that is ethically new when coupled with IVF but requires the same care and concern as AID. Egg or embryo donation, however, does raise new ethical questions that need close attention and continuing analysis. Freezing of human embryos also breaks new ethical ground, particularly in the options it generates beyond a narrowly defined medical domain. Certain of these options are better not undertaken without further public policy decision. Improvement of current procedures and techniques through effective clinical trials can be ethically carried out in terms of scientific and medical perspectives. However, efforts in this direction will be more effective if undertaken within a public policy framework that clearly defines acceptability during a transitional period of confidence-building. PMID:3833444

  8. 牛胚胎移植技术的产业化研究%Study of Industrilization of Bovine Embryo Transferring

    Institute of Scientific and Technical Information of China (English)

    冯建忠; 史远刚; 张秀陶; 梁小军; 薛伟; 扬奇; 孟华; 李毓华

    2001-01-01

    Embryo transfer and relative technology was explicated, including setting up of nucleus herd of cattle by importing frozen embryo, choice of donor, superovalution, cryopreservation of embryo and embryo splitting. This study make a base for the industrilization of bovine embryo transferring.%本文探讨了引进国外优质奶、肉牛冷冻胚胎建立核心群,以及供体牛选择、超数排卵、胚胎冷冻保存和分割等相关技术,为牛胚胎移植产业化奠定基础。

  9. Promising system for selecting healthy in vitro-fertilized embryos in cattle.

    Science.gov (United States)

    Sugimura, Satoshi; Akai, Tomonori; Hashiyada, Yutaka; Somfai, Tamás; Inaba, Yasushi; Hirayama, Muneyuki; Yamanouchi, Tadayuki; Matsuda, Hideo; Kobayashi, Shuji; Aikawa, Yoshio; Ohtake, Masaki; Kobayashi, Eiji; Konishi, Kazuyuki; Imai, Kei

    2012-01-01

    Conventionally, in vitro-fertilized (IVF) bovine embryos are morphologically evaluated at the time of embryo transfer to select those that are likely to establish a pregnancy. This method is, however, subjective and results in unreliable selection. Here we describe a novel selection system for IVF bovine blastocysts for transfer that traces the development of individual embryos with time-lapse cinematography in our developed microwell culture dish and analyzes embryonic metabolism. The system can noninvasively identify prognostic factors that reflect not only blastocyst qualities detected with histological, cytogenetic, and molecular analysis but also viability after transfer. By assessing a combination of identified prognostic factors--(i) timing of the first cleavage; (ii) number of blastomeres at the end of the first cleavage; (iii) presence or absence of multiple fragments at the end of the first cleavage; (iv) number of blastomeres at the onset of lag-phase, which results in temporary developmental arrest during the fourth or fifth cell cycle; and (v) oxygen consumption at the blastocyst stage--pregnancy success could be accurately predicted (78.9%). The conventional method or individual prognostic factors could not accurately predict pregnancy. No newborn calves showed neonatal overgrowth or death. Our results demonstrate that these five predictors and our system could provide objective and reliable selection of healthy IVF bovine embryos. PMID:22590579

  10. Promising system for selecting healthy in vitro-fertilized embryos in cattle.

    Directory of Open Access Journals (Sweden)

    Satoshi Sugimura

    Full Text Available Conventionally, in vitro-fertilized (IVF bovine embryos are morphologically evaluated at the time of embryo transfer to select those that are likely to establish a pregnancy. This method is, however, subjective and results in unreliable selection. Here we describe a novel selection system for IVF bovine blastocysts for transfer that traces the development of individual embryos with time-lapse cinematography in our developed microwell culture dish and analyzes embryonic metabolism. The system can noninvasively identify prognostic factors that reflect not only blastocyst qualities detected with histological, cytogenetic, and molecular analysis but also viability after transfer. By assessing a combination of identified prognostic factors--(i timing of the first cleavage; (ii number of blastomeres at the end of the first cleavage; (iii presence or absence of multiple fragments at the end of the first cleavage; (iv number of blastomeres at the onset of lag-phase, which results in temporary developmental arrest during the fourth or fifth cell cycle; and (v oxygen consumption at the blastocyst stage--pregnancy success could be accurately predicted (78.9%. The conventional method or individual prognostic factors could not accurately predict pregnancy. No newborn calves showed neonatal overgrowth or death. Our results demonstrate that these five predictors and our system could provide objective and reliable selection of healthy IVF bovine embryos.

  11. Bisphenol A affects early bovine embryo development and metabolism that is negated by an oestrogen receptor inhibitor

    Science.gov (United States)

    Choi, Bom-Ie; Harvey, Alexandra J.; Green, Mark P.

    2016-01-01

    Increasing evidence supports an association between exposure to endocrine disruptors, such as the xenoestrogen bisphenol A (BPA), a commonly used plasticiser, and the developmental programming of offspring health. To date however animal studies to investigate a direct causal have mainly focussed on supra-environmental BPA concentrations, without investigating the effect on the early embryo. In this study we investigated the effect of acute BPA exposure (days 3.5 to 7.5 post-fertilisation) at environmentally relevant concentrations (1 and 10 ng/mL) on in vitro bovine embryo development, quality and metabolism. We then examined whether culturing embryos in the presence of the oestrogen receptor inhibitor fulvestrant could negate effects of BPA and 17β-oestradiol (E2). Exposure to BPA or E2 (10 ng/mL) decreased blastocyst rate and the percentage of transferrable quality embryos, without affecting cell number, lineage allocation or metabolic gene expression compared to untreated embryos. Notably, blastocysts exposed to BPA and E2 (10 ng/mL) displayed an increase in glucose consumption. The presence of fulvestrant however negated the adverse developmental and metabolic effects, suggesting BPA elicits its effects via oestrogen-mediated pathways. This study demonstrates that even acute exposure to an environmentally relevant BPA concentration can affect early embryo development and metabolism. These may have long-term health consequences on an individual. PMID:27384909

  12. Bisphenol A affects early bovine embryo development and metabolism that is negated by an oestrogen receptor inhibitor.

    Science.gov (United States)

    Choi, Bom-Ie; Harvey, Alexandra J; Green, Mark P

    2016-01-01

    Increasing evidence supports an association between exposure to endocrine disruptors, such as the xenoestrogen bisphenol A (BPA), a commonly used plasticiser, and the developmental programming of offspring health. To date however animal studies to investigate a direct causal have mainly focussed on supra-environmental BPA concentrations, without investigating the effect on the early embryo. In this study we investigated the effect of acute BPA exposure (days 3.5 to 7.5 post-fertilisation) at environmentally relevant concentrations (1 and 10 ng/mL) on in vitro bovine embryo development, quality and metabolism. We then examined whether culturing embryos in the presence of the oestrogen receptor inhibitor fulvestrant could negate effects of BPA and 17β-oestradiol (E2). Exposure to BPA or E2 (10 ng/mL) decreased blastocyst rate and the percentage of transferrable quality embryos, without affecting cell number, lineage allocation or metabolic gene expression compared to untreated embryos. Notably, blastocysts exposed to BPA and E2 (10 ng/mL) displayed an increase in glucose consumption. The presence of fulvestrant however negated the adverse developmental and metabolic effects, suggesting BPA elicits its effects via oestrogen-mediated pathways. This study demonstrates that even acute exposure to an environmentally relevant BPA concentration can affect early embryo development and metabolism. These may have long-term health consequences on an individual. PMID:27384909

  13. 透明带激光削薄法对IVF-ET中胚胎种植的影响%Influence of zona pellucida Laser-thinning on embryo implantation in IVF-ET

    Institute of Scientific and Technical Information of China (English)

    吕睿; 白晓红; 宋学茹; 鲁琳琳; 岳天孚

    2011-01-01

    目的 探讨激光透明带削薄法对体外受精-胚胎移植(IVF-ET)胚胎种植的影响.方法 回顾性分析243例接受IVF/ICSI治疗的不孕患者,行透明带激光削薄的129名患者为试验组,未行辅助孵化的114例患者作为对照组.结果 (①试验组和对照组患者的一般临床资料相似,无统计学差异.(②试验组患者的种植率较对照组明显升高(30.4% VS 18.8%,P<0.05);临床妊娠率较对照组明显升高(48.8% VS 34.2%,P<0.05);多胎妊娠率较对照组明显升高(34.9% VS 20.5%,P <0.05).结论 激光透明带削薄可以提高临床妊娠率和胚胎的种植率,为降低胚胎移植数量及日后单胚胎移植的实施提供了有利的支持.

  14. Effects of donor cells on in vitro development of cloned bovine embryos

    Institute of Scientific and Technical Information of China (English)

    Jing Fu; Pengfei Guan; Leiwen Zhao; Hua Li; Shuzhen Huang; Fanyi Zeng; Yitao Zeng

    2008-01-01

    The donor cells from different individuals and with different foreign genes introduced were investigated to determine their effects on the efficiency of somatic cell nuclear transfer (SCNT). The bovine ear fibroblast from different individuals was isolated, cultured, and then transfected with foreign genes to establish the stable cell lines, which were used as donor cells for nuclear transfer. The ooeytes were obtained through ovum pick up operation. After in vitro maturation, the M II phase oocytes were selected as receptors for nuclear transfer.The reconstructed embryos were cultured in vitro and observed at 2 h, 48 h, and 7 days after transfer to assess the rate of fusion using cleaved and blastoeyst as the parameters of SCNT efficiency. The donor cells from different individuals (04036, 06081, 06088, and 06129)had no obvious effect on the fusion and cleaved rate, whereas there was significant difference in the blastocyst rate (P0.05). It was concluded that the genetic background of the donor cells could affect the effi-ciency of SCNT, while the introduction of foreign genes into the donor cells had no obvious effect on the efficiency. This study provides useful information for the SCNT and would benefit in promoting the efficiency.

  15. Predicting IVF outcome

    NARCIS (Netherlands)

    L.L. van Loendersloot

    2013-01-01

    On 25 July 1978 at 11.47 PM Louise Brown was born as the first IVF baby ever. Since its introduction more than 5 million babies have been born worldwide using IVF. In contrast to patients’ perception, IVF does not guarantee success; almost 50% of couples that start with IVF will not achieve a pregna

  16. 未成熟卵体外成熟技术在卵巢高反应患者IVF-ET中的应用%In vitro maturation of human immature oocyte in in vitro fertilization and embryo transfer

    Institute of Scientific and Technical Information of China (English)

    黄绘; 洪焱; 冯玉蓉; 刘敏利; 胡皓睿; 潘世春

    2012-01-01

    目的:探讨未成熟卵体外成熟(IVM)技术在卵巢高反应患者体外受精-胚胎移植(IVF - ET)中的应用价值.方法:在IVF - ET促排治疗中,对双卵巢卵泡数过多,有可能发生卵巢过度刺激综合征(OHSS)或继续治疗可能发生重度OHSS的患者,根据其意愿即刻停药,全部取卵改行IVM治疗12个周期(A组)或取部分小卵泡改行IVM治疗,同时保留部分卵泡继续行IVF - ET常规治疗18个周期(B组).小卵泡体外培养成熟后,通过卵胞浆内单精子注射(ICSI)获得受精卵并行胚胎移植或冷冻.统计分析未成熟卵的成熟率、卵母细胞的受精率、胚胎的发育情况及临床结局.结果:两组30个取卵周期,共获未成熟卵240个,经IVM、ICSI和体外培养后,成熟率、受精率、正常卵裂率及优质胚胎率分别为61.25% (147/240),77.55% (114/147),92.98% (106/114)和29.25%(31/106).A组8例行IVM新鲜胚胎移植(8周期)4例临床妊娠,A、B两组有8例行IVM解冻胚胎移植(9周期)3例临床妊娠,已有3例分娩.A组12例无OHSS发生,促性腺激素用量少于B组,B组18例中3例有OHSS风险而取消胚胎移植.结论:对常规IVF促排周期中卵巢高反应患者及时改行IVM,可以避免周期取消及OHSS的发生,减少促排卵药物的使用量,同时获得较好的临床妊娠率.%Objective ; To explore the clinical application of in vitro oocyte maturation (IVM) technique in in vitro fertilization - embryo transfer (IVF - ET). Methods; In the treatment of IVF - ET, women who canceled the treatment cycle due to high risk of developing ovarian hyperstimulation syndrome ( OHSS) were divided into two groups according to their choices. Women in group A (n = 12) underwent oocyte retrieval guided by transvaginal ultrasound for all small follicles and treated with IVM technique for 12 cycles. Women in group B (n = 18) were performed oocyte retrieval for some small follicles treated with IVM technique and kept on IVF - ET with some

  17. In vitro development of OPU-derived bovine embryos cultured either individually or in groups with the silk protein sericin and the viability of frozen-thawed embryos after transfer.

    Science.gov (United States)

    Isobe, Tomohiro; Ikebata, Yoshihisa; Do, Lanh Thi Kim; Tanihara, Fuminori; Taniguchi, Masayasu; Otoi, Takeshige

    2015-07-01

    The optimization of single-embryo culture conditions is very important, particularly in the in vitro production of bovine embryos using the ovum pick-up (OPU) procedure. The purpose of this study was to examine the development of embryos derived from oocytes obtained by OPU that were cultured either individually or in groups in medium supplemented with or without sericin and to investigate the viability of the frozen-thawed embryos after a direct transfer. When two-cell-stage embryos were cultured either individually or in groups for 7 days in CR1aa medium supplemented with or without 0.5% sericin, the rates of development to blastocysts and freezable blastocysts were significantly lower for the embryos cultured individually without sericin than for the embryos cultured in groups with or without sericin. Moreover, the rate of development to freezable blastocysts of the embryos cultured individually with sericin was significantly higher than that of the embryos cultured without sericin. When the frozen-thawed embryos were transferred directly to recipients, the rates of pregnancy, abortion, stillbirth and normal calving in the recipients were similar among the groups, irrespective of the culture conditions and sericin supplementation. Our findings indicate that supplementation with sericin during embryo culture improves the quality of the embryos cultured individually but not the viability of the frozen-thawed embryos after transfer to recipients. PMID:25488699

  18. Relationship between the Expression of sHLA-G in Follicular Fluid and Embryo Culture Fluid and the IVF-ET Outcome%卵泡液和卵裂期胚胎培养液可溶性人类白细胞抗原G(sHLA-G)的表达与IVF-ET结局的相关性研究

    Institute of Scientific and Technical Information of China (English)

    郝天羽; 崔永魁; 刘晓雁; 唐宁; 王建业

    2013-01-01

    目的:探讨卵泡液和卵裂起胚胎培养液中可溶性人类白细胞抗原G(sHLA-G)与卵裂期胚胎发育以及IVF-ET后临床妊娠率的关系,为临床优选卵母细胞及移植胚胎提供一种有效的非侵入性检测途径.方法:采用酶联免疫吸附试验(ELISA)检测取卵日卵泡液及第3日移植胚胎培养液中sHLA-G的表达及表达量,将40例患者分为sHLA-G表达阳性组(n=15)和sHLA-G表达阴性组(n=25)进行比较分析.结果:40份患者卵泡液sHLA-G阳性率为37.5%(15/40),87份第3日移植胚胎培养液中sHLA-G阳性率为56.32%(49/87),卵泡液和胚胎培养液sHLA-G阳性组与阴性组的获卵数和受精率均无差异(P>0.05),sHLA-G阳性组的卵裂率、第3日胚胎卵裂球数、优质胚胎率、临床妊娠率均明显高于阴性组(P<0.05),其中胚胎培养液sHLA-G阳性组的临床妊娠率显著高于阴性组(P<0.01).结论:sHLA-G在部分卵泡液及早期胚胎培养液中均有表达,反映了胚胎良好的发育和种植潜能,预示着较好的IVF-ET结局,是一种优选卵母细胞和移植胚胎的有效的非侵入性途径.%Objective:To investigate the relationship between expression of soluble human leukocyte antigen G (sHLA-G) in follicular fluid and embryo culture fluid and development of cleavage stage embryos and clinical pregnancy rate of IVF-ET and further to provide an effective non-invasive detection method in optimization of oocytes and embryos for implantation.Methods:sHLA-G molecules expression in follicular fluid and embryo culture fluid cultured on the third day was detected by ELISA.Then all 40 patients were divided into sHLAG molecule positive expression group and sHLA-G molecule negative expression group.Results:sHLA-G positive rate was 37.5% (15/40) in follicular fluid and 56.32% (49/87) in embryo culture fluid.In follicular fluid and culture medium which cultured the third day of embryo,there was no significant difference in the number of oocytes

  19. The psychological impact of IVF failure after two or more cycles of IVF with a mild versus standard treatment strategy

    NARCIS (Netherlands)

    C. de Klerk (Cora); N.S. Macklon (Nick); E.M. Heijnen; M.J.C. Eijkemans (René); B.C.J.M. Fauser (Bart); J. Passchier (Jan); J.A.M. Hunfeld (Joke)

    2007-01-01

    textabstractBackground: Failure of IVF treatment after a number of cycles can be devastating for couples. Although mild IVF strategies reduce the psychological burden of treatment, failure may cause feelings of regret that a more aggressive approach, including the transfer of two embryos, was not em

  20. The psychological impact more cycles of IVF with a treatment strategy of IVF failure after two or mild versus standard

    NARCIS (Netherlands)

    de Klerk, C.; Macklon, N. S.; Heijnen, E. M. E. W.; Eijkemans, M. J. C.; Fauser, B. C. J. M.; Passchier, J.; Hunfeld, J. A. M.

    2007-01-01

    BACKGROUND: Failure of IVF treatment after a number of cycles can be devastating for couples. Although mild IVF strategies reduce the psychological burden of treatment, failure may cause feelings of regret that a more aggressive approach, including the transfer of two embryos, was not employed. In t

  1. Splitting of IVP bovine blastocyst affects morphology and gene expression of resulting demi-embryos during in vitro culture and in vivo elongation.

    Science.gov (United States)

    Velasquez, Alejandra E; Castro, Fidel O; Veraguas, Daniel; Cox, Jose F; Lara, Evelyn; Briones, Mario; Rodriguez-Alvarez, Lleretny

    2016-02-01

    Embryo splitting might be used to increase offspring yield and for molecular analysis of embryo competence. How splitting affects developmental potential of embryos is unknown. This research aimed to study the effect of bovine blastocyst splitting on morphological and gene expression homogeneity of demi-embryos and on embryo competence during elongation. Grade I bovine blastocyst produced in vitro were split into halves and distributed in nine groups (3 × 3 setting according to age and stage before splitting; age: days 7-9; stage: early, expanded and hatched blastocysts). Homogeneity and survival rate in vitro after splitting (12 h, days 10 and 13) and the effect of splitting on embryo development at elongation after embryo transfer (day 17) were assessed morphologically and by RT-qPCR. The genes analysed were OCT4, SOX2, NANOG, CDX2, TP1, TKDP1, EOMES, and BAX. Approximately 90% of split embryos had a well conserved defined inner cell mass (ICM), 70% of the halves had similar size with no differences in gene expression 12 h after splitting. Split embryos cultured further conserved normal and comparable morphology at day 10 of development; this situation changes at day 13 when embryo morphology and gene expression differed markedly among demi-embryos. Split and non-split blastocysts were transferred to recipient cows and were recovered at day 17. Fifty per cent of non-split embryos were larger than 100 mm (33% for split embryos). OCT4, SOX2, TP1 and EOMES levels were down-regulated in elongated embryos derived from split blastocysts. In conclusion, splitting day-8 blastocysts yields homogenous demi-embryos in terms of developmental capability and gene expression, but the initiation of the filamentous stage seems to be affected by the splitting.

  2. Down- regulation of three long protocols on in vitro fertilization and embryo transfer outcome%3种长方案垂体降调节对IVF - ET结局的影响

    Institute of Scientific and Technical Information of China (English)

    江成龙; 郑蓉; 黄晓卉; 罗丽娟; 周晓艳; 胡冬玲; 张昌军

    2012-01-01

    目的:探讨3种长方案垂体降调节对体外受精-胚胎移植( IVF - ET)结局的影响.方法:752个IVF - ET周期患者分为A、B、C3组,均行长方案降调节控制超促排卵.A组304周期应用短效促性腺激素释放激素激动剂( GnRH - a)0.05 mg/d;B组218周期应用短效GnRH -a 0.1 mg/d注射10~ 14d后改为0.05mg/d;C组230周期应用长效GnRH -a 1.25mg单次注射.比较3组促性腺激素(Gn)使用剂量、使用时间及IVF - ET结局.结果:降调时间(开始降调至Gn启动时间)B组长于C组(P<0.05),C组长于A组(P <0.05);Gn使用时间C组长于B组(P<0.05),B组长于A组(P <0.05);Gn使用总量、受精率C组多于和高于A、B两组(P<0.05);优胚率B组高于A、C组(P<0.05);临床妊娠率B、C两组差异无统计学意义(P>0.05),但均高于A组(P<0.05);中、重度卵巢过度刺激综合征(OHSS)发生率、多胎率、流产率3组间差异无统计学意义(P>0.05).结论:增加GnRHa的使用剂量,延缓Gn启动时间可以改善IVF - ET结局,但会相应增加Gn使用剂量,增加患者治疗费用.长效GnRHa l.25mg单次注射也可得到满意的降调节效果和临床妊娠率.%Objective: To explore effects of three kinds of down - regulation protocols on clinical outcomes of in vitro fertilization and embryo transfer (IVF - ET). Methods: A total of 752 women were enrolled and randomized into 3 groups prospec-tively. Women received 0.05 mg/d of short - acting gonadotropin - releasing hormone agonist ( GnRHa) injection in group A (304 cycles). Women in group B (218 cycles) were administered 0. 1 mg/d GnRHa for 10 to 14 days , followed by 0. 05 mg/d until human chorionic gonadotrophin (hCG) administration. In group C (230 cycles) , women were injected 1.25 mg of long - acting GnRHa. The mean days and dose of gonadotrophin use as well as the clinical outcome of IVF - ET were compared among the three groups. Results: The mean down - regulation day in group B was

  3. Developmental kinetics of the first cell cycles of bovine in vitro PRODUCED EMBRYOS IN RELATION TO THEIR IN VITRO VIABILITY AND SEX

    DEFF Research Database (Denmark)

    Holm, P; Shukri, N.N; Vajta, Gabor;

    1998-01-01

    The development of bovine IVP-embryos was observed in a time-lapse culture system to determine cell cycle lengths of 1) embryos that developed into compact morulae (CM) or blastocysts (BL) within 174 h after insemination (viable), 2) embryos that arrested during earlier stages (nonviable) and 3...... were included (n=392), and the times of cleavage events noted. After culture, 100 CM or BL were randomly selected for sexing by PCR. BL developed equally well in the time-lapse and control culture systems (36 vs 38. The respective lengths of the first 4 cell cycles of viable embryos were 32.0 + 3.9, g...

  4. Birth of live calves by in vitro embryo production of slaughtered cows in a commercial herd in South Africa

    Directory of Open Access Journals (Sweden)

    T. Arlotto

    2001-07-01

    Full Text Available In vitro fertilisation (IVF has become a useful breeding tool in most of the developed world. In this paper the success of bovine IVF and the birth of live calves under typical South African conditions is reported. Oocytes for IVF were collected from the ovaries of 6 slaughtered Bovelder beef cows. On average, 36.2 oocytes per donor were retrieved. From these oocytes, 43 blastocysts were produced from 5 of the donors by IVF with frozen Bovelder semen. The best 11 of these embryos were transferred into oestrous, synchronised Bovelder recipients in the same herd. As a result, 7 calves were born (a 64 %calving rate from 4 of the original donors. The calves had a normal birth mass, but the mean gestation length of the male calves was significantly longer than the herd average (291.6 versus 285.2 days respectively. No calving difficulties were encountered. In summary, it was shown that IVF for bovine embryo production and transfer is possible on a commercial basis in South Africa.

  5. Derivation of HVR1, HVR2 and HVR3 human embryonic stem cell lines from IVF embryos after preimplantation genetic diagnosis (PGD for monogenic disorder

    Directory of Open Access Journals (Sweden)

    Abdelkrim Hmadcha

    2016-05-01

    Full Text Available From 106 human blastocyts donate for research after in vitro fertilization (IVF and preimplantation genetic diagnosis (PGD for monogenetic disorder, 3 human embryonic stem cells (hESCs HVR1, HVR2 and HVR3 were successfully derived. HVR1 was assumed to be genetically normal, HVR2 carrying Becker muscular dystrophy and HVR3 Hemophilia B. Despite the translocation t(9;15(q34.3;q14 detected in HVR2, all the 3 cell lines were characterised in vitro and in vivo as normal hESCs lines and were registered in the Spanish Stem Cell Bank.

  6. Derivation of HVR1, HVR2 and HVR3 human embryonic stem cell lines from IVF embryos after preimplantation genetic diagnosis (PGD) for monogenic disorder.

    Science.gov (United States)

    Hmadcha, Abdelkrim; Aguilera, Yolanda; Lozano-Arana, Maria Dolores; Mellado, Nuria; Sánchez, Javier; Moya, Cristina; Sánchez-Palazón, Luis; Palacios, Jose; Antiñolo, Guillermo; Soria, Bernat

    2016-05-01

    From 106 human blastocyts donate for research after in vitro fertilization (IVF) and preimplantation genetic diagnosis (PGD) for monogenetic disorder, 3 human embryonic stem cells (hESCs) HVR1, HVR2 and HVR3 were successfully derived. HVR1 was assumed to be genetically normal, HVR2 carrying Becker muscular dystrophy and HVR3 Hemophilia B. Despite the translocation t(9;15)(q34.3;q14) detected in HVR2, all the 3 cell lines were characterised in vitro and in vivo as normal hESCs lines and were registered in the Spanish Stem Cell Bank. PMID:27346196

  7. Effect of zona pellucida removal on early development of in vitro produced bovine embryos Efecto de la remoción de la zona pelúcida sobre el desarrollo temprano de embriones bovinos producidos in vitro

    Directory of Open Access Journals (Sweden)

    AE Velásquez

    2013-01-01

    Full Text Available During early embryo development the zona pellucida acts as a barrier against polyspermia and guarantees communication between blastomeres before and during compaction. However, the development of new technologies of embryo production such as "Handmade Cloning" demands removal of this membrane. The aim of this study was to determine the effect of zona pellucida removal on in vitro bovine embryo development. First, the consequences of zona pellucida removal was assessed by comparing the percentage of first cleavage, percentage of blastocysts and cell number among zona-included and zona-free embryos; either through the removal of the zona pellucida immediately after IVF or parthenogenesis. Embryo development was also evaluated when zona pellucida was removed before parthenogenesis. In a second set of experiments, the gene expression levels of BAX, BCL2, CASP3, CDH1, OCT4 and SOX2 were evaluated in zona-free and zona-included IVF-derived embryos. No significant differences were found in the percentage of first cleavage, percentage of blastocyst and cell number on IVF-embryos cultured with or without zona. Parthenogenetic embryos followed the same general pattern even when the zona pellucida was eliminated before activation; however there was a significant increase in the rate of first cleavage when the zona pellucida was removed after activation but this did not impact upon further development. Furthermore, no significant differences in gene expression level were found between zona-free and zona-included IVF-embryos for the studied genes. We concluded that the lack of zona pellucida did not affect the early development when an appropriate system is used for embryo culture to ensure blastomeric contact and normal compaction.Durante el desarrollo embrionario temprano, la zona pelúcida actúa como barrera contra la polispermia y además garantiza la comunicación entre blastómeras y la compactación de estas. Sin embargo, el desarrollo de nuevas

  8. Effect of insulin-like growth factor-1 during in vitro oocyte maturation and in vitro culture of bovine embryos

    OpenAIRE

    Quetglas M.D.; Coelho L.A.; Garcia J.M.; Oliveira Filho E.B.; Esper C.R.

    2001-01-01

    The effects of insulin-like growth factor-I (IGF-I) on in vitro maturation (IVM) (experiment I) and on in vitro embryo development (experiment II) of bovine oocytes fertilized in vitro, were evaluated in terms of cleavage (CR), blastocyst (BR) and hatching (HR) rates. For IVM, immature cumulus-oocyte complexes were cultured in TCM-199 medium supplemented with Hepes, sodium bicarbonate, sodium pyruvate, additives, fetal calf serum (B-199 medium) and gonadotropins (14 U/ml PMSG and 7 U/ml hCG)....

  9. Kinetics of fertilization and development, and sex ratio of bovine embryos produced using the semen of different bulls.

    Science.gov (United States)

    Alomar, M; Tasiaux, H; Remacle, S; George, F; Paul, D; Donnay, I

    2008-08-01

    The between bulls variation in in vitro fertility and the shift of sex ratio towards male embryos are two problems affecting the in vitro production (IVP) of bovine embryos. Our objective was to evaluate the kinetics of fertilization, embryo development and the sex ratio of the resulting embryos using the frozen/thawed semen of four different bulls. In a first experiment, the kinetics of pronucleus (PN) formation was evaluated at 8, 12 and 18 h post-insemination (hpi). Based upon the pronuclei sizes and the distance between the two pronuclei, inseminated oocytes were classified in three PN stages. Differences between bulls were observed at each time point, but were more important at 12 hpi. At 8 and 12 hpi bull III showed a significantly faster PN evolution by comparison with the three other bulls (Pcinematography. The analysis of embryos reaching the blastocyst stage revealed significant differences in the mean time of first cleavage (range of 22.7-25.6h, P<0.05), while the lengths of the subsequent three cell cycles did not differ between bulls. The early mean time of first cleavage with bull III was associated with an early blastulation and a high blastocyst rate at Day 7, in opposition to what was observed with bull II showing a later timing of first cleavage (first cleavage 22.1 hpi versus 25.5 hpi; blastulation 140.4 hpi versus 152.5 hpi; D7 blastocyst rates: 31.3% versus 21.9%; P<0.05). In a third experiment, 65-76 Day 8 blastocysts per bull were sexed by PCR. Only blastocysts obtained with bull III showed a shift in sex ratio towards male embryos (76% male embryos; P<0.05). Such shift was already observed at the 2-cell and morula stages. In conclusion, the bull influences the kinetics of PN formation, of embryo development and the sex ratio of the embryos. Moreover, those parameters might be related. PMID:17629423

  10. Effects of elective double embryo transfer on pregnancy outcome in IVF-ET%选择性双胚胎移植对IVF-ET妊娠结局的影响

    Institute of Scientific and Technical Information of China (English)

    董胜男; 刘炜; 王炎秋; 周围; 石礼红; 赵起越; 谢晓东; 纪亚忠

    2015-01-01

    目的 探讨体外授精-胚胎移植(IVF-ET)中,选择性双胚胎移植对妊娠结局的影响. 方法 收集在上海市同济医院生殖中心从2012年6月至2014年5月行两枚第3天胚胎移植的517个周期,按照移植胚胎的形态学评分分为三组:A组为移植两枚Ⅰ级胚胎(304个周期);B组移植一枚Ⅰ级胚胎和一枚低级别的胚胎(128个周期);C组为移植两枚非Ⅰ级胚胎(85个周期).分别对各组间的患者年龄、不育年限、不育因素以及临床妊娠率、抱婴率、活产率等临床结局进行统计学比较. 结果 3组每移植周期的临床妊娠率和抱婴率依次降低,分别为73.7%、62.5%、43.5%和62.1%、51.6%、36.5%,组间均有统计学差异(P<0.05);3组间的异位妊娠率、早期流产率、晚期流产率和活产率差异均无统计学意义(P>0.05);C组的生化妊娠率显著高于A、B两组(P<0.05),A、B组间差异不显著(P>0.05);A组的胚胎活产率显著高于B、C两组(P<0.05),B、C组间差异不显著(P>0.05). 结论 胚胎形态学分级可以作为评判胚胎发育潜能的参数,Ⅰ级胚胎可获得最好的临床妊娠率和胚胎活产率;胚胎成功种植后,胚胎形态学分级不影响妊娠结局.

  11. In vitro fertilization (IVF)

    Science.gov (United States)

    IVF; Assisted reproductive technology; ART; Test-tube baby procedure; Infertility - in vitro ... This process is called natural or unassisted conception. IVF is a form of assisted reproductive technology (ART). ...

  12. IVF and retinoblastoma revisited.

    NARCIS (Netherlands)

    Dommering, C.J.; Hout, A.H. van der; Meijers-Heijboer, H.; Marees, T.; Moll, A.C.

    2012-01-01

    OBJECTIVE: To evaluate the suggested association between IVF, retinoblastoma, and tumor methylation characteristics. DESIGN: Laboratory analysis. SETTING: National Retinoblastoma Center in the Netherlands. PATIENT(S): Retinoblastoma tumors from seven children conceived by IVF or intracytoplasmic spe

  13. IVF and retinoblastoma revisited

    NARCIS (Netherlands)

    Dommering, Charlotte J.; van der Hout, Annemarie H.; Meijers-Heijboer, Hanne; Marees, Tamara; Moll, Annette C.

    2012-01-01

    Objective: To evaluate the suggested association between IVF, retinoblastoma, and tumor methylation characteristics. Design: Laboratory analysis. Setting: National Retinoblastoma Center in the Netherlands. Patient(s): Retinoblastoma tumors from seven children conceived by IVF or intracytoplasmic spe

  14. Effect of Equilibration Temperature on In vitro Viability and Subsequent Embryo Development of Vitrified-Warmed Immature Bovine Oocytes

    Directory of Open Access Journals (Sweden)

    Hajarian Hadi

    2010-01-01

    Full Text Available Problem statement: Vitrification is replacing conventional slow freezing to cryopreserve gametes and embryos especially for in vitro production of embryo in domestic animal species. However, the results are still not satisfactory. The aim of this experiment was to study the effect of different equilibration temperatures on in vitro viability of immature bovine oocytes after vitrification. Approach: Oocytes were obtained from slaughterhouse ovaries. Only grade one oocytes were used. Oocytes were equilibrated in three different temperatures: 32, 37, or 41°C. Immature oocytes were equilibrated in VS1 (7.5 Ethylene Glycol (EG + 7.5% DMSO for 10-12 min and then exposed to VS2 (15% EG + 15%DMSO + 0.5M sucrose for one min. Thereafter oocytes were loaded on hand-made Cryotop and directly plunged into liquid nitrogen. After warming, oocytes were examined for viability, maturation, cleavage and blastocyst production. Results: Oocytes that were equilibrated at 37°C had significantly higher (pConclusion: In conclusion, these results indicated that immature bovine oocytes can be equilibrated successfully at 37°C while higher or lower temperature can significantly decrease their subsequent viability and development.

  15. Finding biomarkers in non-model species: literature mining of transcription factors involved in bovine embryo development

    Directory of Open Access Journals (Sweden)

    Turenne Nicolas

    2012-08-01

    Full Text Available Abstract Background Since processes in well-known model organisms have specific features different from those in Bos taurus, the organism under study, a good way to describe gene regulation in ruminant embryos would be a species-specific consideration of closely related species to cattle, sheep and pig. However, as highlighted by a recent report, gene dictionaries in pig are smaller than in cattle, bringing a risk to reduce the gene resources to be mined (and so for sheep dictionaries. Bioinformatics approaches that allow an integration of available information on gene function in model organisms, taking into account their specificity, are thus needed. Besides these closely related and biologically relevant species, there is indeed much more knowledge of (i trophoblast proliferation and differentiation or (ii embryogenesis in human and mouse species, which provides opportunities for reconstructing proliferation and/or differentiation processes in other mammalian embryos, including ruminants. The necessary knowledge can be obtained partly from (i stem cell or cancer research to supply useful information on molecular agents or molecular interactions at work in cell proliferation and (ii mouse embryogenesis to supply useful information on embryo differentiation. However, the total number of publications for all these topics and species is great and their manual processing would be tedious and time consuming. This is why we used text mining for automated text analysis and automated knowledge extraction. To evaluate the quality of this “mining”, we took advantage of studies that reported gene expression profiles during the elongation of bovine embryos and defined a list of transcription factors (or TF, n = 64 that we used as biological “gold standard”. When successful, the “mining” approach would identify them all, as well as novel ones. Methods To gain knowledge on molecular-genetic regulations in a non model organism, we offer an

  16. Screening of biotechnical parameters for production of bovine inter-subspecies embryonic chimeras by the aggregation of tetraploid Bos indicus and diploid crossbred Bos taurus embryos.

    Science.gov (United States)

    Razza, Eduardo M; Satrapa, Rafael A; Emanuelli, Isabele P; Barros, Ciro M; Nogueira, Marcelo F G

    2016-03-01

    The aggregation of a tetraploid zebu embryo (Bos indicus, a thermotolerant breed) with a diploid taurine embryo (Bos taurus, a thermosensitive breed) should create a complete taurine fetus, whose extra-embryonic components, e.g., the chorion, is derived mainly from the zebu embryo. These zebu-derived extra-embryonic components may interact positively with the taurine embryo/fetus during pregnancy in a tropical environment. We tested different parameters for the production of tetraploid Nelore (Bos indicus) embryos to be combined via aggregation with crossbred Bos taurus (diploid) embryos in order to produce viable chimeric blastocysts. Bovine (Bos indicus or crossbred Bos taurus) embryos were produced in vitro according to standard procedures. Two-cell Bos indicus embryos were submitted to electrofusion with varying numbers of pulses (1 or 2), voltages (0.4, 0.5, 0.75, 1.0, 1.4 and 5.0 kV/cm) and time (20, 25, 50 and 60 μs) to produce tetraploid embryos. Electrofused embryos were cultured with crossbred non-fused embryos to form chimeras that developed until the blastocyst stage. The best fusion parameter was 0.75 kV/cm for 60 μs. Four chimeric blastocysts (tetraploid Nelore with diploid crossbred Holstein) were formed after 31 attempts in 4 replicates (13%). We established an optimal procedure for the production of tetraploid Bos indicus (4n) embryos and embryonic chimeras by aggregation of crossbred Bos taurus (2n) with Bos indicus (4n) embryos. This technique would be valid in applied research, by producing exclusively taurine calves, but with placental elements from the Bos indicus breed, following transfer of these chimeras into recipient cows.

  17. Effect of cryopreservation and in vitro culture of bovine fibroblasts on histone acetylation levels and in vitro development of hand-made cloned embryos

    Science.gov (United States)

    Chacon, L.; Gomez, M.C.; Jenkins, J.A.; Leibo, S.P.; Wirtu, G.; Dresser, B.L.; Pope, C.E.

    2011-01-01

    In this study, the relative acetylation levels of histone 3 in lysine 9 (H3K9ac) in cultured and cryopreserved bovine fibroblasts was measured and we determined the influence of the epigenetic status of three cultured (C1, C2 and C3) donor cell lines on the in vitro development of reconstructed bovine embryos. Results showed that cryopreservation did not alter the overall acetylation levels of H3K9 in bovine fibroblasts analysed immediately after thawing (frozen/thawed) compared with fibroblasts cultured for a period of time after thawing. However, reduced cleavage rates were noted in embryos reconstructed with fibroblasts used immediately after thawing. Cell passage affects the levels of H3K9ac in bovine fibroblasts, decreasing after P1 and donor cells with lower H3K9ac produced a greater frequency of embryo development to the blastocyst stage. Cryopreservation did not influence the total cell and ICM numbers, or the ICM/TPD ratios of reconstructed embryos. However, the genetic source of donor cells did influence the total number of cells and the trophectoderm cell numbers, and the cell passage influenced the total ICM cell numbers. ?? Copyright Cambridge University Press 2010.

  18. Gene expression, oocyte nuclear maturation and developmental competence of bovine oocytes and embryos produced after in vivo and in vitro heat shock.

    Science.gov (United States)

    Pavani, Krishna C; Baron, Erica; Correia, Pedro; Lourenço, Joana; Bettencourt, Bruno Filipe; Sousa, Madalena; da Silva, Fernando Moreira

    2016-10-01

    Three assays were performed. In assay 1, oocytes harvested during the winter months were subjected to kinetic heat shock by stressing the oocytes at 39.5°C (HS1) or at 40.5°C (HS2) for either 6, 12, 18 or 24 h and then matured at control temperature (38.5°C). The nuclear maturation rates (NMR) of all oocytes were recorded after 24 h. In assay 2, oocytes collected year-round maturated, were implanted via in vitro fertilization (IVF) and developed for 9 days. Gene expression analysis was performed on target genes (Cx43, CDH1, DNMT1, HSPA14) with reference to the two housekeeping genes (GAPDH and SDHA) in embryos. Similarly, in assay 3, genetic analysis was performed on the embryos produced from heat-stressed oocytes (from HS1 and HS2). In assay 1, the duration of heat stress resulted in a significant decline in NMR (P CDH1 genes (P < 0.05). Targeted gene expression was aberrant in embryo development, which can provide evidence on early embryo arrest and slowed embryo development.

  19. cDNA microarray analysis of bovine embryo gene expression profiles during the pre-implantation period

    Directory of Open Access Journals (Sweden)

    Tokunaga Tomoyuki

    2004-11-01

    Full Text Available Abstract Background After fertilization, embryo development involves differentiation, as well as development of the fetal body and extra-embryonic tissues until the moment of implantation. During this period various cellular and molecular changes take place with a genetic origin, e.g. the elongation of embryonic tissues, cell-cell contact between the mother and the embryo and placentation. To identify genetic profiles and search for new candidate molecules involved during this period, embryonic gene expression was analyzed with a custom designed utero-placental complementary DNA (cDNA microarray. Methods Bovine embryos on days 7, 14 and 21, extra-embryonic membranes on day 28 and fetuses on days 28 were collected to represent early embryo, elongating embryo, pre-implantation embryo, post-implantation extra-embryonic membrane and fetus, respectively. Gene expression at these different time points was analyzed using our cDNA microarray. Two clustering algorithms such as k-means and hierarchical clustering methods identified the expression patterns of differentially expressed genes across pre-implantation period. Novel candidate genes were confirmed by real-time RT-PCR. Results In total, 1,773 individual genes were analyzed by complete k-means clustering. Comparison of day 7 and day 14 revealed most genes increased during this period, and a small number of genes exhibiting altered expression decreased as gestation progressed. Clustering analysis demonstrated that trophoblast-cell-specific molecules such as placental lactogens (PLs, prolactin-related proteins (PRPs, interferon-tau, and adhesion molecules apparently all play pivotal roles in the preparation needed for implantation, since their expression was remarkably enhanced during the pre-implantation period. The hierarchical clustering analysis and RT-PCR data revealed new functional roles for certain known genes (dickkopf-1, NPM, etc as well as novel candidate genes (AW464053, AW465434, AW

  20. Combination of IVF and IVM in naturally cycling women

    DEFF Research Database (Denmark)

    Tang-Pedersen, Mikael; Westergaard, Lars Grabow; Erb, Karin;

    2012-01-01

    This study investigated the combination of an unstimulated IVF cycle with in-vitro maturation (IVM) of additional immature cumulus-oocyte-complexes (COC) from the same cycle collected at the same time as the spontaneous preovulatory follicle. This could potentially improve rates of embryo transfer...... and pregnancy/live births compared with conventional unstimulated IVF treatment and at the same time eliminate the risk of ovarian hyperstimulation syndrome. This prospective trial included 77 women with regular menstrual cycles. Age at inclusion was between 20 and 37 years. Results showed a retrieval rate...... between endometrial factors and IVM oocytes together with unknown competence of IVM embryos is suspected. For some time, there has been an increasing interest in mild approaches for fertility treatment, in particular IVF. In-vitro maturation (IVM) of immature eggs outside the ovaries followed by IVF...

  1. Characteristics of pregnancies and offspring following transfer of bovine in vivo embryos assessed by nanorespirometry

    DEFF Research Database (Denmark)

    Lopes, Ana Sofia; Madsen, S E; Greve, Torben;

    2008-01-01

    It has been speculated whether the metabolism of the pre-implantation embryo may be reflected on the pregnancy and characteristics of the newborn animal. The present study investigated whether respiration rates of individual embryos were correlated with gestation length, type of parturition, birth......, III), stage of development, and diameter and were subsequently transferred individually (n = 43) to synchronized recipients. Gestation length of the recipients (n = 22) was calculated and the type of parturition (no assistance, light traction, heavy traction, or caesarean section) recorded. Sex......, weight, and condition of the calves at birth (weak, normal, or very active) were also assessed. Results were evaluated by chi-square analysis and using a linear mixed model. The pregnancy rate was 60% (26/43), and the respiration rates of individual embryos influenced gestation length as well...

  2. Invited review: Genetic contributions underlying the development of preimplantation bovine embryos.

    Science.gov (United States)

    Kropp, J; Peñagaricano, F; Salih, S M; Khatib, H

    2014-03-01

    In recent years, it has become evident that genetic selection to improve milk production has resulted in a decline in dairy cattle fertility. Growing evidence suggests that the greatest loss occurs early in pregnancy around the time of embryo implantation. As a means to make genetic improvements and to assist in reproductive performance, use of artificial reproductive technologies such as artificial insemination and in vitro production of embryos have been widely used. Both of these technologies rely on the competence and quality of gametes for successful development of embryos. Often, selection of animals is based on the genetic merit of the animal, although specific fertility markers are relatively underdeveloped compared with markers for production traits. Similarly, current in vitro fertilization systems could benefit from a uniform method for selection of the best quality embryos to transfer into recipients for successful implantation and delivery of healthy offspring. As genetics underlie biological processes such as fertility, the need exists to further identify and characterize genes that affect fertility and development within both the parental gametes and the embryo. Furthermore, the magnitude of the contribution of each parental genome to the success of embryo development and pregnancy is not clear. As such, the objective of this review is to provide an overview of studies relating to genetic markers at the DNA level, parental and embryonic gene expression, and the effects of epigenetics on embryonic development. Future studies should exploit advances in molecular technologies to identify and classify genes underlying fertility and development to establish biomarkers and predictors for improved genetic selection. PMID:24377798

  3. Production of bovine cloned embryos with donor cells frozen at a slow cooling rate in a conventional freezer (20 C)

    Science.gov (United States)

    Chacon, L.; Gomez, M.C.; Jenkins, J.A.; Leibo, S.P.; Wirtu, G.; Dresser, B.L.; Pope, C.E.

    2009-01-01

    Summary Usually, fibroblasts are frozen in dimethyl sulphoxide (DMSO, 10% v/v) at a cooling rate of 1 C/min in a low-temperature (80 C) freezer (LTF) before storage in liquid nitrogen (LN2); however, a LTF is not always available. The purpose of the present study was to evaluate apoptosis and viability of bovine fibroblasts frozen in a LTF or conventional freezer (CF; 20 C) and their subsequent ability for development to blastocyst stage after fusion with enucleated bovine oocytes. Percentages of live cells frozen in LTF (49.5%) and CF (50.6%) were similar, but significantly less than non-frozen control (88%). In both CF and LTF, percentages of live apoptotic cells exposed to LN2 after freezing were lower (4% and 5%, respectively) as compared with unexposed cells (10% and 18%, respectively). Cells frozen in a CF had fewer cell doublings/24 h (0.45) and required more days (9.1) to reach 100% confluence at the first passage (P) after thawing and plating as compared with cells frozen in a LTF (0.96 and 4.0 days, respectively). Hypoploidy at P12 was higher than at P4 in cells frozen in either a CF (37.5% vs. 19.2%) or in a LTF (30.0% vs. 15.4%). A second-generation cryo-solution reduced the incidence of necrosis (29.4%) at 0 h after thawing as compared with that of a first generation cryo-solution (DMEM + DMSO, 60.2%). The percentage of apoptosis in live cells was affected by cooling rate (CF = 1.9% vs. LFT = 0.7%). Development of bovine cloned embryos to the blastocyst stage was not affected by cooling rate or freezer type. ?? 2009 Cambridge University Press.

  4. Simulated Microgravity Influences Bovine Oocyte In Vitro Fertilization and Preimplantation Embryo Development

    Science.gov (United States)

    The aim of this study was to investigate whether in vitro fertilization and preimplantation embryos exposed to a simulated microgravity environment in vitro would improve, or be deleterious to, their fertilization and embryonic development. A Rotating Cell Culture System™ (RCCS) bioreactor with a Hi...

  5. Influence of recipient cytoplasm cell stage on transcription in bovine nucleus transfer embryos

    DEFF Research Database (Denmark)

    Smith, Steven D.; Soloy, Eva; Kanka, Jiri;

    1996-01-01

    . NTE were produced using either a MII phase (nonactivated) cytoplasts at 32 hr of maturation or S-phase (activated) cytoplasts activated with calcium ionophore A23187 and cycloheximide treatment approximately 8 hr prior to fusion with a blastomere from an in-vitro-produced morula stage embryo at 32 hr...

  6. Expression of intracellular interferon-alpha confers antiviral properties in transfected bovine fetal fibroblasts and does not affect the full development of SCNT embryos.

    Directory of Open Access Journals (Sweden)

    Dawei Yu

    Full Text Available Foot-and-mouth disease, one of the most significant diseases of dairy herds, has substantial effects on farm economics, and currently, disease control measures are limited. In this study, we constructed a vector with a human interferon-α (hIFN-α (without secretory signal sequence gene cassette containing the immediate early promoter of human cytomegalovirus. Stably transfected bovine fetal fibroblasts were obtained by G418 selection, and hIFN-α transgenic embryos were produced by somatic cell nuclear transfer (SCNT. Forty-six transgenic embryos were transplanted into surrogate cows, and five cows (10.9% became pregnant. Two male cloned calves were born. Expression of hIFN-α was detected in transfected bovine fetal fibroblasts, transgenic SCNT embryos, and different tissues from a transgenic SCNT calf at two days old. In transfected bovine fetal fibroblasts, expression of intracellular IFN-α induced resistance to vesicular stomatitis virus infection, increased apoptosis, and induced the expression of double-stranded RNA-activated protein kinase gene (PKR and the 2'-5'-oligoadenylate synthetase gene (2'-5' OAS, which are IFN-inducible genes with antiviral activity. Analysis by qRT-PCR showed that the mRNA expression levels of PKR, 2'-5' OAS, and P53 were significantly increased in wild-type bovine fetal fibroblasts stimulated with extracellular recombinant human IFN-α-2b, showing that intracellular IFN-α induces biological functions similar to extracellular IFN-α. In conclusion, expression of intracellular hIFN-α conferred antiviral properties in transfected bovine fetal fibroblasts and did not significantly affect the full development of SCNT embryos. Thus, IFN-α transgenic technology may provide a revolutionary way to achieve elite breeding of livestock.

  7. Transcriptional reprogramming of gene expression in bovine somatic cell chromatin transfer embryos

    OpenAIRE

    Page Grier P; Kasinathan Poothappillai; Wang Zhongde; Rodriguez-Osorio Nelida; Robl James M; Memili Erdogan

    2009-01-01

    Abstract Background Successful reprogramming of a somatic genome to produce a healthy clone by somatic cells nuclear transfer (SCNT) is a rare event and the mechanisms involved in this process are poorly defined. When serial or successive rounds of cloning are performed, blastocyst and full term development rates decline even further with the increasing rounds of cloning. Identifying the "cumulative errors" could reveal the epigenetic reprogramming blocks in animal cloning. Results Bovine clo...

  8. Transcriptional Reprogramming of Gene Expression in Bovine Somatic Cell Chromatin Transfer Embryos

    OpenAIRE

    Rodriguez-Osorio, N.; Wang, Zhongde; Page, G. P.; Robl, J M; Memili, E.

    2009-01-01

    Background Successful reprogramming of a somatic genome to produce a healthy clone by somatic cells nuclear transfer (SCNT) is a rare event and the mechanisms involved in this process are poorly defined. When serial or successive rounds of cloning are performed, blastocyst and full term development rates decline even further with the increasing rounds of cloning. Identifying the "cumulative errors" could reveal the epigenetic reprogramming blocks in animal cloning. Results Bovine clones from ...

  9. Effects of Antioxidants on Development of In Vitro Fertilized Bovine Embryos

    OpenAIRE

    Anderson, Bret L.

    1995-01-01

    Free radicals are short-lived molecules that can cause decreased embryonic development in vitro. Antioxidants are molecules that block free radical formation or guard against their harmful effects. Many studies have linked exposure of media to light and culturing of embryos in high (20%) oxygen concentrations to free radical production. Some of the antioxidants used in culture media are superoxide dismutase (SOD), catalase, zinc (II), ethylenedinitrilo tetraacetic acid (EDTA), mannitol, vitam...

  10. Selection of reference genes for quantitative real-time PCR in bovine preimplantation embryos

    Directory of Open Access Journals (Sweden)

    Van Zeveren Alex

    2005-12-01

    Full Text Available Abstract Background Real-time quantitative PCR is a sensitive and very efficient technique to examine gene transcription patterns in preimplantation embryos, in order to gain information about embryo development and to optimize assisted reproductive technologies. Critical to the succesful application of real-time PCR is careful assay design, reaction optimization and validation to maximize sensitivity and accuracy. In most of the studies published GAPD, ACTB or 18S rRNA have been used as a single reference gene without prior verification of their expression stability. Normalization of the data using unstable controls can result in erroneous conclusions, especially when only one reference gene is used. Results In this study the transcription levels of 8 commonly used reference genes (ACTB, GAPD, Histone H2A, TBP, HPRT1, SDHA, YWHAZ and 18S rRNA were determined at different preimplantation stages (2-cell, 8-cell, blastocyst and hatched blastocyst in order to select the most stable genes to normalize quantitative data within different preimplantation embryo stages. Conclusion Using the geNorm application YWHAZ, GAPD and SDHA were found to be the most stable genes across the examined embryonic stages, while the commonly used ACTB was shown to be highly regulated. We recommend the use of the geometric mean of those 3 reference genes as an accurate normalization factor, which allows small expression differences to be reliably measured.

  11. Examination of Early Cleavage an its Importance in IVF Treatment

    Directory of Open Access Journals (Sweden)

    Fancsovits P

    2006-01-01

    Full Text Available Since the introduction of assisted reproduction, the number of multiple pregnancies has increased due to the high number of transferred embryos. There is an urgent need for IVF specialists to reduce the number of embryos transferred without the risk of decreasing pregnancy rates. Embryos are selected for transfer on the basis of their developmental stage and morphology. The number of blastomeres of the embryo indicates the speed of early embryo development which correlates to the viability of the embryo. Examination of early embryo development, especially the timing of the first cleavage, can be recommended as a tool for predicting embryo viability. Observation of timing of the first cleavage and its different stages helps us to identify fast- and slow-developing embryos. Early pronuclear breakdown and early cleavage of the zygote are indicators of fast embryo development and good embryo viability. Thereby, they can lead to high implantation and pregnancy rates. The aim of this paper is to provide an overview of the timing of early embryo development and to show its significance in IVF treatment.

  12. In vitro fertilization (IVF) in mammals: epigenetic and developmental alterations. Scientific and bioethical implications for IVF in humans.

    Science.gov (United States)

    Ventura-Juncá, Patricio; Irarrázaval, Isabel; Rolle, Augusto J; Gutiérrez, Juan I; Moreno, Ricardo D; Santos, Manuel J

    2015-12-18

    The advent of in vitro fertilization (IVF) in animals and humans implies an extraordinary change in the environment where the beginning of a new organism takes place. In mammals fertilization occurs in the maternal oviduct, where there are unique conditions for guaranteeing the encounter of the gametes and the first stages of development of the embryo and thus its future. During this period a major epigenetic reprogramming takes place that is crucial for the normal fate of the embryo. This epigenetic reprogramming is very vulnerable to changes in environmental conditions such as the ones implied in IVF, including in vitro culture, nutrition, light, temperature, oxygen tension, embryo-maternal signaling, and the general absence of protection against foreign elements that could affect the stability of this process. The objective of this review is to update the impact of the various conditions inherent in the use of IVF on the epigenetic profile and outcomes of mammalian embryos, including superovulation, IVF technique, embryo culture and manipulation and absence of embryo-maternal signaling. It also covers the possible transgenerational inheritance of the epigenetic alterations associated with assisted reproductive technologies (ART), including its phenotypic consequences as is in the case of the large offspring syndrome (LOS). Finally, the important scientific and bioethical implications of the results found in animals are discussed in terms of the ART in humans.

  13. Intrafallopian transfer of gametes and early stage embryos for in vivo culture in cattle.

    Science.gov (United States)

    Wetscher, F; Havlicek, V; Huber, T; Gilles, M; Tesfaye, D; Griese, J; Wimmers, K; Schellander, K; Müller, M; Brem, G; Besenfelder, U

    2005-07-01

    It may be possible to avoid inadequate in vitro culture conditions by incubating gametes or embryos in the oviducts for a short time. Ideally, an optimized procedure should be devised, combining in vitro and in vivo systems, in order to achieve synchronization in cattle. We transferred gametes as well as embryos in various stages of development and placed them into the oviducts. Embryos were recovered on Day 7 by flushing of oviducts and uterine horns. Blastocyst rates were determined on Day 7 and on Day 8. Experimental designs included transfer of in vitro matured cumulus oocyte complexes into previously inseminated heifers (COCs group), transfer of in vitro matured COCs simultaneously with capacitated spermatozoa (GIFTs group), transfer of four to eight cell stage embryos developed in vitro after IVM/IVF (Cleaved Stages group) and a group of solely in vitro produced embryos (IVP control group). Our results indicate that in vivo culture of IVM/IVF embryos in the homologous bovine oviduct has a positive influence on subsequent pre-implantation development. In addition, we have evidence that in vitro maturation and in vivo fertilization cannot be synchronized. PMID:15935840

  14. Genetic variation in resistance of the preimplantation bovine embryo to heat shock.

    Science.gov (United States)

    Hansen, Peter J

    2014-12-01

    Reproduction is among the physiological functions in mammals most susceptible to disruption by hyperthermia. Many of the effects of heat stress on function of the oocyte and embryo involve direct effects of elevated temperature (i.e. heat shock) on cellular function. Mammals limit the effects of heat shock by tightly regulating body temperature. This ability is genetically controlled: lines of domestic animals have been developed with superior ability to regulate body temperature during heat stress. Through experimentation in cattle, it is also evident that there is genetic variation in the resistance of cells to the deleterious effects of elevated temperature. Several breeds that were developed in hot climates, including Bos indicus (Brahman, Gir, Nelore and Sahiwal) and Bos taurus (Romosinuano and Senepol) are more resistant to the effects of elevated temperature on cellular function than breeds that evolved in cooler climates (Angus, Holstein and Jersey). Genetic differences are expressed in the preimplantation embryo by Day 4-5 of development (after embryonic genome activation). It is not clear whether genetic differences are expressed in cells in which transcription is repressed (oocytes >100 µm in diameter or embryos at stages before embryonic genome activation). The molecular basis for cellular thermotolerance has also not been established, although there is some suggestion for involvement of heat shock protein 90 and the insulin-like growth factor 1 system. Given the availability of genomic tools for genetic selection, identification of genes controlling cellular resistance to elevated temperature could be followed by progress in selection for those genes within the populations in which they exist. It could also be possible to introduce genes from thermotolerant breeds into thermally sensitive breeds. The ability to edit the genome makes it possible to design new genes that confer protection of cells from stresses like heat shock. PMID:25472041

  15. Daily supplementation with ghrelin improves in vitro bovine blastocysts formation rate and alters gene expression related to embryo quality.

    Science.gov (United States)

    Dovolou, Eleni; Periquesta, Eva; Messinis, Ioannis E; Tsiligianni, Theodora; Dafopoulos, Konstantinos; Gutierrez-Adan, Alfonso; Amiridis, Georgios S

    2014-03-01

    Ghrelin is a gastric peptide having regulatory role in the reproductive system functionality, acting mainly at central level. Because the expression of ghrelin system (ghrelin and its receptor) has been detected in the bovine ovary, the objectives of the present study were to investigate whether ghrelin can affect the developmental potential of in vitro-produced embryos, and to test their quality in terms of relative abundance of various genes related to metabolism, apoptosis and oxidation. In the first experiment, in vitro-produced zygotes were cultured in the absence (control [C]) and in the presence of three concentrations of acylated ghrelin (200 pg/mL [Ghr200], 800 pg/mL [Ghr800]; and 2000 pg/mL [Ghr2000]); blastocyst formation rates were examined on Days 7, 8, and 9. In the second experiment, only the 800 pg/mL dose of ghrelin was used. Zygotes were produced as in experiment 1 and 24 hours post insemination they were divided into 4 groups; in two groups (C; without ghrelin; Ghr800 with ghrelin), embryos were cultured without medium replacement; in the remaining two groups (Control N and GhrN), the culture medium was daily renewed. A pool of Day-7 blastocysts were snap frozen for relative mRNA abundance of various genes related to metabolism, oxidation, implantation, and apoptosis. In experiment 3, embryos were produced as in experiment 2, but in the absence of serum (semi-defined culture medium). In experiment 1, no differences were detected between C, Ghr200, and Ghr2000, although fewer blastocysts were produced in group Ghr800 compared with C. In experiment 2, the lowest blastocysts yield was found in Ghr800, whereas daily renewal of ghrelin (Ghr800N) resulted to increased blastocysts formation rate, which on Day 7 was the highest among groups (P quality than controls. Our results imply a specific role of ghrelin in early embryonic development; however, the specific mode of its action needs further investigation. PMID:24332928

  16. Gestaciones producidas con embriones bovinos clonados por transferencia nuclear Pregnancies produced by bovine embryos cloned by nuclear transfer

    Directory of Open Access Journals (Sweden)

    M A Martínez Díaz

    2007-01-01

    Full Text Available El presente estudio comunica la obtención de gestaciones de embriones clonados por transferencia nuclear de células somáticas en bovinos por primera vez en Chile. Ovocitos bovinos obtenidos de ovarios de matadero fueron madurados in vitro y enucleados por micromanipulación. Células donantes de núcleos fueron obtenidas de la oreja de una vaca adulta, cultivadas por 9-14 días y criopreservadas en nitrógeno líquido. Células somáticas confluentes fueron desagregadas e insertadas individualmente en el espacio perivitelino de un ovocito enucleado. Cada par ovocito-célula obtenido fue tratado con dos pulsos eléctricos para inducir su fusión y luego los embriones fueron cultivados por 2 horas y tratados con ionomicina y 6-dimetilaminopurina para su activación. Los embriones fueron cultivados en medio sintético oviductual por 8-9 días hasta el estado de blastocisto. Blastocistos fueron transferidos a vaquillas receptoras 7 a 9 días postcelo. Se realizaron 25 transferencias a vaquillas receptoras y se logró la preñez en 5 (20% de ellas. Dos de éstas abortaron a los 42 días y una tercera a los 120 días. Las dos vaquillas preñadas restantes mantuvieron su gestación (más de 45 días hasta el momento de escribir esta comunicación.With the aim of obtaining pregnancies from nuclear transfer embryos reconstituted with somatic cells and enucleated oocytes, bovine oocytes from slaughterhouse were matured in vitro and enucleated by micromanipulation. Nuclear donor cells were obtained from the ear of an adult cow, cultured for 9 to 14 days and cryopreserved in liquid nitrogen. Confluent cells were inserted individually in the perivitelline space of enucleated oocytes and treated with 2 electric pulses to induce fusion. The reconstituted zygotes were then cultured for 2 hours and treated with ionomycin and 6-dimethylaminopurine for their activation. The embryos were cultured in synthetic oviduct fluid for 8 to 9 days to obtain the

  17. MODELAGEM BIOECONÔMICA DA TRANSFERÊNCIA DE EMBRIÕES EM BOVINOS BIOECONOMIC MODEL IN BOVINE EMBRYO TRANSFER

    Directory of Open Access Journals (Sweden)

    Renato Travassos Beltrame

    2010-04-01

    Full Text Available

    O objetivo deste trabalho foi desenvolver um modelo matemático orientado a eventos de simulação, para auxiliar tomadas de decisão relativas à transferência de embriões em bovinos, considerando-se as dinâmicas de dois componentes da transferência de embriões: receptoras e embriões. Na simulação, não se avaliaram respostas individuais de doadoras a coletas consecutivas e eventos correspondentes na transferência de embriões. Simulou-se o mesmo protocolo para superovulação a todas as doadoras. Receptoras foram sincronizadas simulando-se o uso de prostaglandina. O número de embriões viáveis produzido por doadora e sua variabilidade tiveram como base um processo aleatório de simulação de Monte Carlo, que pressupôs uma distribuição exponencial negativa de densidade de probabilidade. Custos e receitas foram inseridos no modelo por meio de um cenário-base para calcular indicadores econômicos de rentabilidade. A análise sugeriu a impraticabilidade da atividade, se realizada diante do cenário proposto (VPL – R$: 57.596,69. A partir do cenário proposto, o custo médio estimado foi de R$ 1.178,19, e de R$ 980,03, para se obter uma prenhez a partir de uma situação otimizada, sugerida pelo modelo (5/100; 5/190.

    PALAVRAS-CHAVES: Otimização, receptoras, simulação, transferência de embriões, viabilidade econômica.

    A simulation model related to embryo transfer programs in bovine was carried out through a mathematical model directed to events, considering the dynamic of two resources: recipients and embryos. Individual answers of donors to consecutive collections and corresponding events in embryo transfer were not evaluated. The same protocol for superovulation was simulated for all the donor collections, using similar doses of hormones and drugs for all the animals. Recipients were synchronized using prostaglandin. Meantime, the number of viable embryos produced by donor and its variability were based at

  18. Influence of L-arginine during bovine in vitro fertilization.

    Science.gov (United States)

    Silva, Thiago Velasco Guimarães; da Silva, Bruno Baraúna; de Sá, André Luiz Alves; da Costa, Nathalia Nogueira; Sampaio, Rafael Vilar; Cordeiro, Marcela da Silva; Santana, Priscila Di Paula Bessa; Adona, Paulo Roberto; Santos, Simone do Socorro Damasceno; Miranda, Moysés dos Santos; Ohashi, Otávio Mitio

    2014-12-01

    The objective of this work was to evaluate the effect of using L-arginine during in vitro fertilization (IVF) on in vitro embryonic development using Bos taurus and Bos indicus semen. Effect of different concentrations (0, 1, 10 and 50 mM) of L-arginine, added to the IVF medium, was evaluated on the fertilization rate at 18 h post-fertilization (hpf), NO3(-)/NO2(-) production during IVF by the Griess colorimetric method (30 hpf), cleavage and blastocyst rates (on Day 2 and Day 7 of culture, respectively) and total blastocyst cell number (Day 7 of culture). The results reveal that the addition of 50 mM L-arginine to IVF medium, with either Bos taurus or Bos indicus spermatozoa, decreased the cleavage rate and blastocyst rate compared to the control group. Other concentrations did not affect embryo production. However, 1 mM L-arginine with Bos indicus semen increased the proportion of hatched blastocysts. These results indicate that high L-arginine concentrations may exhibit toxic effects on bovine gametes during in vitro fertilization. PMID:25651608

  19. Effect of zona pellucida removal on early development of in vitro produced bovine embryos Efecto de la remoción de la zona pelúcida sobre el desarrollo temprano de embriones bovinos producidos in vitro

    OpenAIRE

    AE Velásquez; JR Manríquez; FO Castro; Ll Rodríguez-Alvarez

    2013-01-01

    During early embryo development the zona pellucida acts as a barrier against polyspermia and guarantees communication between blastomeres before and during compaction. However, the development of new technologies of embryo production such as "Handmade Cloning" demands removal of this membrane. The aim of this study was to determine the effect of zona pellucida removal on in vitro bovine embryo development. First, the consequences of zona pellucida removal was assessed by comparing the percent...

  20. Embryo cloning by nuclear transfer in the bovine species: first results

    OpenAIRE

    Ectors, Francis; Ectors, Fabien; Delval, Alain; Thonon, Fabienne; Beckers, Jean-François

    1993-01-01

    Le clonage par transfert de noyau fut réalisé pour la première fois chez les bovins par Prather et collaborateurs en 1987. Vu l'importance économique de ce mode de multiplication, de nombreuses équipes de recherche affinent la technique et étudient sa mise en application sur le terrain. Alors que l'énucléation de l'ovocyte receveur, l'injection et la fusion du blastomère donneur se réalisent avec succès, la maturation ovocytaire, la culture et la congélation de l'embryon reconstitué posent en...

  1. Embryo splitting: a role in infertility?

    Science.gov (United States)

    Wood, C

    2001-01-01

    Embryo splitting may be used to increase the potential fertility of couples requiring IVF. Using cattle as a model, it is possible to increase pregnancy rates from 70% per transfer of good quality in-vivo-produced embryos, to 110% by transferring the two demi-embryos resulting from the bisection of one embryo. The 30-40% greater chance of conception would reduce costs for the government, health authorities and patients, and reduce stress, time and complications for women having IVF treatment. Embryo splitting may also provide donor embryos for infertile couples that cannot conceive naturally or with IVF. The shortage of children for adoption and donor embryos may be overcome by the production of demi-embryos.

  2. Aromatase Inhibitors for IVF Poor Responders

    Institute of Scientific and Technical Information of China (English)

    R.B. Quintero; L.C Giudice; L.M. Westphal

    2006-01-01

    Objective To evaluate whether letrozole enhanced follicular recruitment, embryo numbers, and pregnancy rates in poor responders undergoing IVF.Methods We reviewed all IVF cycles between January 2002 and September 2003 using letrozole at Stanford University Medical Center. The entry criteria were the requirement of at least 450 IU/d of injectable gonadotropins in a prior failed cycle,which was used as a control.Results A total of 27 charts were reviewed revealing information on 54 cycles. The number of oocytes retrieved, fertilization embryo quality and embryos transferred yielded no statistical significance, although there appeared to be a trend toward higher numbers of each in the letrozole group. The clinical pregnancy rate was 9/27 (33.3%, P<0. 001)with a viable pregnancy rate of 7/27 (25.9%, P=0.002) in the letrozole cycle.Conclusion Our study is one of the first to evaluate letrozole with in vitro fertilization.Although this study showed no difference in number of oocytes or embryos, 25.9% of these "poor responding" patients achieved a pregnancy after a failed cycle at our center.

  3. Regulation of pluripotency of inner cell mass and growth and differentiation of trophectoderm of the bovine embryo by colony stimulating factor 2.

    Science.gov (United States)

    Dobbs, Kyle B; Khan, Firdous A; Sakatani, Miki; Moss, James I; Ozawa, Manabu; Ealy, Alan D; Hansen, Peter J

    2013-12-01

    Colony-stimulating factor 2 (CSF2) enhances competence of the bovine embryo to establish and maintain pregnancy after the embryo is transferred into a recipient. Mechanisms involved could include regulation of lineage commitment, growth, or differentiation of the inner cell mass (ICM) and trophectoderm (TE). Experiments were conducted to evaluate regulation by CSF2 of pluripotency of the ICM and differentiation and growth of the TE. Embryos were cultured with 10 ng/ml recombinant bovine CSF2 or a vehicle control from Days 5 to 7 or 6 to 8 postinsemination. CSF2 increased the number of putative zygotes that developed to blastocysts when the percent of embryos becoming blastocysts in the control group was low but decreased blastocyst yield when blastocyst development in controls was high. ICM isolated from blastocysts by lysing the trophectoderm using antibody and complement via immunosurgery were more likely to survive passage when cultured on mitomycin C-treated fetal fibroblasts if derived from blastocysts treated with CSF2 than if from control blastocysts. There was little effect of CSF2 on characteristics of TE outgrowths from blastocysts. The exception was a decrease in outgrowth size for embryos treated with CSF2 from Days 5 to 7 and an increase in expression of CDX2 when treatment was from Days 6 to 8. Expression of the receptor subunit gene CSF2RA increased from the zygote stage to the 9-16 cell stage before decreasing to the blastocyst stage. In contrast, CSF2RB was undetectable at all stages. In conclusion, CSF2 improves competence of the ICM to survive in a pluripotent state and alters TE outgrowths. Actions of CSF2 occur through a signaling pathway that is likely to be independent of CSF2RB.

  4. The ISMAAR proposal on terminology for ovarian stimulation for IVF

    NARCIS (Netherlands)

    Nargund, G.; Fauser, B. C. J. M.; Macklon, N. S.; Ombelet, W.; Nygren, K.; Frydman, R.

    2007-01-01

    IVF is performed with oocytes collected in natural and stimulated cycles. Different approaches to ovarian stimulation have been employed worldwide. Following the introduction of GnRH antagonists and strategies to reduce multiple births such as single embryo transfer, there is a genuine scientific in

  5. Use of versapoint to refashion the cervical canal to overcome unusually difficult embryo transfers and improve in-vitro fertilization-embryo transfer outcome: A case series

    OpenAIRE

    Nalini Mahajan; Ila Gupta

    2011-01-01

    Background : Smooth atraumatic embryo transfer is paramount for the success of in-vitro fertilization (IVF). In difficult cases, cervical canal manipulation may be required. Aim : To see if surgical correction of the cervical canal or cervical canal refashioning could improve ease of embryo transfer. Setting : Private infertility and IVF hospital. Design : Prospective study. Materials and Methods : Patients: 11 women with failed 1-3 IVF cycles with history of extremely difficult embryo transf...

  6. Time-lapse cinematography-compatible polystyrene-based microwell culture system: a novel tool for tracking the development of individual bovine embryos.

    Science.gov (United States)

    Sugimura, Satoshi; Akai, Tomonori; Somfai, Tamás; Hirayama, Muneyuki; Aikawa, Yoshio; Ohtake, Masaki; Hattori, Hideshi; Kobayashi, Shuji; Hashiyada, Yutaka; Konishi, Kazuyuki; Imai, Kei

    2010-12-01

    We have developed a polystyrene-based well-of-the-well (WOW) system using injection molding to track individual embryos throughout culture using time-lapse cinematography (TLC). WOW culture of bovine embryos following in vitro fertilization was compared with conventional droplet culture (control). No differences between control- and WOW-cultured embryos were observed during development to the blastocyst stage. Morphological quality and inner cell mass (ICM) and trophectoderm (TE) cell numbers were not different between control- and WOW-derived blastocysts; however, apoptosis in both the ICM and TE cells was reduced in WOW culture (P < 0.01). Oxygen consumption in WOW-derived blastocysts was closer to physiological level than that of control-derived blastocysts. Moreover, WOW culture improved embryo viability, as indicated by increased pregnancy rates at Days 30 and 60 after embryo transfer (P < 0.05). TLC monitoring was performed to evaluate the cleavage pattern and the duration of the first cell cycle of embryos from oocytes collected by ovum pickup; correlations with success of pregnancy were determined. Logistic regression analysis indicated that the cleavage pattern correlated with success of pregnancy (P < 0.05), but cell cycle length did not. Higher pregnancy rates (66.7%) were observed for animals in which transferred blastocysts had undergone normal cleavage, identified by the presence of two blastomeres of the same size without fragmentation, than among those with abnormal cleavage (33.3%). These results suggest that our microwell culture system is a powerful tool for producing and selecting healthy embryos and for identifying viability biomarkers. PMID:20739661

  7. Study on Sex Determination of Bovine Pre-implantation Embryos By Bovine Y Chromosome Repeated Sequence%利用牛Y染色体重复序列进行早期胚胎性别鉴定的研究

    Institute of Scientific and Technical Information of China (English)

    王世银; 张伟; 张兆旺; 赵兴绪

    2011-01-01

    本试验利用Y染色体重复序列作为雄性特异性引物,以肿瘤坏死因子(TNF-α) 内标引物建立多重PCR体系,进行牛早期胚胎性别鉴定.共设计四对引物一Y染色体重复序列外引物和内引物,其大小分别为534bp和480bp;肿瘤坏死因子外引物和内引物大小分别为357bp和272bp.试验结果表明,优化后的多重PCR体系的灵敏度分别达到3个胚胎细胞,准确率100%,可以满足早期胚胎性别鉴定的需要.%In this study, we designed four pairs of primers which the amplifiment products length were 534bp, 480bp, 357bp and 272bp respectively according to Y chromosome repeated sequence and tumor necrosis factor alpha(TNF-α) for sex determination of bovine embryo.The result shows that these four pairs of primers all have highly specificity and stability.The Multi-PCR need only 3 cells DNA to determine the sex of embryo, so it is more suitable for sex determination of bovine embryo.

  8. Active caspase-3 and ultrastructural evidence of apoptosis in spontaneous and induced cell death in bovine in vitro produced pre-implantation embryos

    DEFF Research Database (Denmark)

    Gjørret, Jakob O.; Fabian, Dusan; Avery, Birthe;

    2007-01-01

    In this study we investigated chronological onset and involvement of active caspase-3, apoptotic nuclear morphology, and TUNEL-labeling, as well as ultrastructural evidence of apoptosis, in both spontaneous and induced cell death during pre-implantation development of bovine in vitro produced...... staining for detection of apoptotic nuclear morphology, and subjected to fluorescence microscopy. Additionally, treated and untreated blastocysts were fixed and processed for ultrastructural identification of apoptosis. Untreated embryos revealed no apoptotic features at 2- and 4-cell stages. However......, active caspase-3 and apoptotic nuclear morphology were observed in an untreated 8-cell stage, and TUNEL-labeling was observed from the 16-cell stage. Blastomeres concurrently displaying all apoptotic features were present in a few embryos at 16-cell and morula stages and in all blastocysts. All three...

  9. Improved preimplantation development of bovine ICSI embryos generated with spermatozoa pretreated with membrane-destabilizing agents lysolecithin and Triton X-100.

    Science.gov (United States)

    Zambrano, Fabiola; Aguila, Luis; Arias, María E; Sánchez, Raúl; Felmer, Ricardo

    2016-10-01

    In cattle, intracytoplasmic sperm injection (ICSI) has a low efficiency. The acrosome content may be responsible for this effect because of the large amount of hydrolytic enzymes that are released within the oocyte. With the aim of removing the acrosome and destabilize the membranes, cryopreserved bovine spermatozoa were treated with lysolecithin (LL) and Triton X-100 (TX) at different concentrations. We evaluated the membrane integrity, the acrosome integrity, DNA integrity, and the variation of phospholipase C zeta. The rates of development (cleavage and blastocysts) were also evaluated along with pronuclear formation and the embryo quality. Spermatozoa incubated with LL and TX (0.01%, 0.02%, 0.03%, and 0.04%) decreased (P embryonic development, without affecting the quality of the embryos produced by this technique. PMID:27325573

  10. Omics in Reproductive Medicine: Application of Novel Technologies to Improve the IVF Success Rate.

    Science.gov (United States)

    Nerenz, R D

    2016-01-01

    Treatment for many infertile couples often consists of in vitro fertilization (IVF) but an estimated 70% of IVF cycles fail to produce a live birth. In an attempt to improve the live birth rate, the vast majority of IVF cycles performed in the United States involve the transfer of multiple embryos, a practice that increases the risk of multiple gestation pregnancy. This is a concern because multiple gestation pregnancies are associated with an increased incidence of maternal and fetal complications and significant cost associated with the care of preterm infants. As the ideal outcome of each IVF cycle is the birth of a single healthy baby, significant effort has focused on identifying embryos with the greatest developmental potential. To date, selection of euploid embryos using comprehensive chromosome screening (CCS) is the most promising approach while metabolomic and proteomic assessment of spent culture medium have the potential to noninvasively assess embryo viability. Endometrial gene expression profiling may help determine the optimal time to perform embryo transfer. While CCS has been implemented in some clinics, further development and optimization will be required before analysis of spent culture medium and endometrial gene expression profiling make the transition to clinical use. This review will describe efforts to identify embryos with the greatest potential to result in a healthy, live birth, with a particular emphasis on detection of embryo aneuploidy and metabolic profiling of spent embryo culture medium. Assessment of endometrial receptivity to identify the optimal time to perform embryo transfer will also be discussed. PMID:27645816

  11. Is IVF-served two different ways-more cost-effective than IUI with controlled ovarian hyperstimulation?

    NARCIS (Netherlands)

    Tjon-Kon-Fat, R. I.; Bensdorp, A. J.; Bossuyt, P. M M; Koks, C.; Oosterhuis, G. J E; Hoek, A.; Hompes, P.; Broekmans, F. J.; Verhoeve, H. R.; De Bruin, J. P.; Van Golde, R.; Repping, S.; Cohlen, B. J.; Lambers, M. D A; Van Bommel, P. F.; Slappendel, E.; Perquin, D.; Smeenk, J.; Pelinck, M. J.; Gianotten, J.; Hoozemans, D. A.; Maas, J. W M; Groen, H.; Eijkemans, M. J C; Van Der Veen, F.; Mol, B. W J; Van Wely, M.

    2015-01-01

    STUDY QUESTION What is the cost-effectiveness of in vitro fertilization (IVF) with conventional ovarian stimulation, single embryo transfer (SET) and subsequent cryocycles or IVF in a modified natural cycle (MNC) compared with intrauterine insemination with controlled ovarian hyperstimulation (IUI-C

  12. Effects of Two Types of Melatonin-Loaded Nanocapsules with Distinct Supramolecular Structures: Polymeric (NC) and Lipid-Core Nanocapsules (LNC) on Bovine Embryo Culture Model

    Science.gov (United States)

    Komninou, Eliza Rossi; Remião, Mariana Härter; Lucas, Caroline Gomes; Domingues, William Borges; Basso, Andrea Cristina; Jornada, Denise Soledade; Deschamps, João Carlos; Beck, Ruy Carlos Ruver; Pohlmann, Adriana Raffin; Bordignon, Vilceu; Seixas, Fabiana Kömmling; Campos, Vinicius Farias; Guterres, Silvia Stanisçuaski; Collares, Tiago

    2016-01-01

    Melatonin has been used as a supplement in culture medium to improve the efficiency of in vitro produced mammalian embryos. Through its ability to scavenge toxic oxygen derivatives and regulate cellular mRNA levels for antioxidant enzymes, this molecule has been shown to play a protective role against damage by free radicals, to which in vitro cultured embryos are exposed during early development. In vivo and in vitro studies have been performed showing that the use of nanocapsules as active substances carriers increases stability, bioavailability and biodistribution of drugs, such as melatonin, to the cells and tissues, improving their antioxidant properties. These properties can be modulated through the manipulation of formula composition, especially in relation to the supramolecular structures of the nanocapsule core and the surface area that greatly influences drug release mechanisms in biological environments. This study aimed to evaluate the effects of two types of melatonin-loaded nanocapsules with distinct supramolecular structures, polymeric (NC) and lipid-core (LNC) nanocapsules, on in vitro cultured bovine embryos. Embryonic development, apoptosis, reactive oxygen species (ROS) production, and mRNA levels of genes involved in cell apoptosis, ROS and cell pluripotency were evaluated after supplementation of culture medium with non-encapsulated melatonin (Mel), melatonin-loaded polymeric nanocapsules (Mel-NC) and melatonin-loaded lipid-core nanocapsules (Mel-LNC) at 10−6, 10−9, and 10−12 M drug concentrations. The highest hatching rate was observed in embryos treated with 10−9 M Mel-LNC. When compared to Mel and Mel-NC treatments at the same concentration (10−9 M), Mel-LNC increased embryo cell number, decreased cell apoptosis and ROS levels, down-regulated mRNA levels of BAX, CASP3, and SHC1 genes, and up-regulated mRNA levels of CAT and SOD2 genes. These findings indicate that nanoencapsulation with LNC increases the protective effects of

  13. First successful pregnancies following embryo selection using Time-lapse technology in Iran: Case report

    OpenAIRE

    Azita Faramarzi; Mohammad Ali Khalili; Mehrdad Soleimani

    2015-01-01

    Background: Embryo selection is a vital part of in vitro fertilization (IVF) programs, with morphology-based grading systems having been widely used for decades. Time-lapse imaging combined with embryo morph kinetics may proffer a non-invasive means for improving embryo selection. We report the first ongoing and chemical pregnancies using Time-lapse embryo scope to select best embryos for transfer in Iran. Cases: A case with tubal factor infertility was admitted to IVF program with normozo...

  14. Axiomatic Characterizations of IVF Rough Approximation Operators

    Directory of Open Access Journals (Sweden)

    Guangji Yu

    2014-01-01

    Full Text Available This paper is devoted to the study of axiomatic characterizations of IVF rough approximation operators. IVF approximation spaces are investigated. The fact that different IVF operators satisfy some axioms to guarantee the existence of different types of IVF relations which produce the same operators is proved and then IVF rough approximation operators are characterized by axioms.

  15. The near elimination of triplets in IVF.

    Science.gov (United States)

    Gerris, Jan

    2007-01-01

    In Antwerp, single embryo transfers (SET) have increased, and this change in policy has seen the incidence of singletons rise from 70 to 90%, twins drop from 25 to 10%, and triplets drop from 1-2% to none at all. At a national and regional level, changes have been much more dramatic as a result of a change in the law in 2003 mandating SET. Data show a huge increase in the number of treatment cycles and a dramatic rise in SET. Rates of twins and triplets have dropped considerably. European data indicate wide differences in the incidence of triplets between countries, varying between 0% (e.g.Slovenia, Iceland, Lithuania) and 4.4% (Hungary), but almost nothing is known about the true incidence of fetal reduction. US data indicate some decrease in triplets during recent years, but certainly nowhere near elimination. These data suggest that in some countries, IVF triplets have almost been eliminated, but the problem is masked by significant fetal reductions. Where SET is not widely used, triplets are still frequent. A correlation appears to exist between the percentage of SET cycles and the incidence of triplets. It is suggested that the solution to almost totally eliminate triplets after IVF is single embryo transfer.

  16. Patients with polycystic ovary syndrome have successful embryo arrest

    OpenAIRE

    Yin, Baoli; Hao, Haoying; Wei, Duo; Song, Xiaobing; Xie, Juanke; Zhang, Cuilian

    2015-01-01

    In this retrospective study, we investigate the relationship between embryo arrest and polycystic ovary syndrome (PCOS) during in vitro fertilization-embryo transfer (IVF-ET). In this study, 667 subjects were enrolled, including 330 patients with PCOS and 337 subjects without PCOS. The subjects underwent in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET) cycles at the Reproductive Medical Centre of Henan Provincial Hospital from January 2009 to December ...

  17. Post-hatching development of in vitro bovine embryos from day 7 to 14 in vivo versus in vitro.

    Science.gov (United States)

    Machado, G M; Ferreira, A R; Pivato, I; Fidelis, A; Spricigo, J F; Paulini, F; Lucci, C M; Franco, M M; Dode, M A

    2013-11-01

    This study evaluates the post-hatching development of in vitro-produced (IVP) embryos until Day 14. On Day 7, IVP embryos were either transferred to recipient uteruses or placed in a post-hatching development (PHD) system. As a control group, in vivo-produced (IVV), Day-7 embryos were also transferred to recipient uteruses. All groups were collected on Day 14 and were morphologically evaluated. Day-7 and Day-14 IVV and IVP embryos were used for quantification of eight genes (PLAC8, CD9, SLC2A1, SLC2A3, KRT8, SOD2, HSP1A1, and IFNT2) by reverse transcriptase qPCR. Day-14 embryos from the PHD system were smaller (2.92 ± 0.45 mm) and had a lower embryonic disk diameter (0.14 ± 0.00 mm) than those produced by IVV (24.18 ± 3.71; 0.29 ± 0.03 mm, respectively) or IVP (19.06 ± 2.43; 0.28 ± 0.01 mm) culture and transferred to the uterus (P > 0.05). Day-7 IVP embryos had a higher expression of the HSP1A1, SCL2A1, and SCL2A3 genes than IVV embryos. When these embryos were cultured in the uterus, no differences in gene expression were observed on Day 14. Conversely, Day-14 IVP embryos cultured in the PHD system showed a higher expression of PLAC8, SOD2, and SLC2A3 genes. It is concluded that Day-7 IVP embryos are different from IVV embryos in regards to gene expression, although exposure to the uterine environment during the elongation period allowed the IVP embryos to overcome this difference. In contrast, IVP embryos cultured in the PHD system were morphologically and molecularly different, being of poorer quality than those cultured in the uterus.

  18. Lessons from elective in vitro fertilization (IVF in, principally, non-infertile women

    Directory of Open Access Journals (Sweden)

    Gleicher Norbert

    2012-06-01

    Full Text Available Abstract Background We here report the first investigation of exclusively elective in vitro fertilization (IVF cycles in women with no apparent history of infertility. Since IVF outcome in women with infertility are always influenced by underlying causes of infertility, a study on non-infertile women may offer new insights. Methods We investigated 88 females without history of infertility in 109 consecutive elective IVF cycles, almost exclusively performed for purposes of preimplantation genetic screening (PGS; i.e., elective gender selection. The following questions were addressed: (i impact of PGS on IVF pregnancy chances; (ii impact of transfer of 1 vs. ≥2 embryos on IVF pregnancy chances; (iii correlation of anti-Müllerian hormone (AMH levels to embryo ploidy (iv effect of gonadotropin dosage used in stimulation on available embryos for transfer; and (v in form of a 1:1 case control study, compared 33 elective PGS cycles with matched control cycles without PGS, performed in couples with either prior tubal ligations and/or severe male factor infertility as indication of IVF. Results The overall clinical pregnancy rate for the group was 36.7%; pregnancy was associated with number of euploid (P = 0.009 and number of embryos transferred (P = 0.001. Odds of pregnancy were 3.4-times higher if ≥4 euploid embryos were produced in comparison to Conclusions This study suggests that outcomes of elective IVF cycles may significantly deviate from infertility-associated cycles. Affirming proof of concept for PGS, utilizing day-3 embryo biopsy and fluorescence in-situ hybridization (FISH, both widely held responsible for earlier failures to establish such proof, suggests that the principal cause of prior failures were likely not insufficient laboratory techniques but poor patient selection for PGS. Such a conclusion questions the current reintroduction of PGS with improved techniques and technologies in absence of prior determination of suited

  19. Improvements in IVF in women of advanced age.

    Science.gov (United States)

    Gleicher, Norbert; Kushnir, Vitaly A; Albertini, David F; Barad, David H

    2016-07-01

    Women above age 40 years in the US now represent the most rapidly growing age group having children. Patients undergoing in vitro fertilization (IVF) are rapidly aging in parallel. Especially where egg donations are legal, donation cycles, therefore, multiply more rapidly than autologous IVF cycles. The donor oocytes, however, are hardly ever a preferred patient choice. Since with use of own eggs, live birth rates decline with advancing age but remain stable (and higher) with donor eggs, older patients always face the difficult and very personal choice between poorer chances with own and better chances with donor oocytes. Physician contribution to this decision should in our opinion be restricted to accurate outcome information for both options. Achievable pregnancy and live birth rates in older women are, however, frequently underestimated, thereby mistakenly biasing fertility providers, private insurance companies and even regulatory government agencies. Restriction on access to IVF for older women is then often the consequence. In this review, we summarize the limited published data on best treatments of 'older' ovaries, while also addressing treatment approaches that should be avoided in older women. This focused review, therefore, to a degree is subjective. Research addressing aging ovaries in IVF has been disappointingly sparse, and has in our opinion too heavily concentrated on methods of embryo selection (ES), which, especially in older women, not only fail to improve IVF outcomes, but actually, negatively affect live birth chances. We conclude that, aside from breakthroughs in gamete creation, only pharmacological interventions into early (small growing follicle stages) follicle maturation will offer new potential to positively impact oocyte and embryo quality and, therefore, IVF outcomes. Research, therefore, should be accordingly redirected.

  20. 基础卵泡刺激素水平和获卵数对IVF-ET结局的影响%Impact of serum FSH level and the number of retrieved oocytes on clinical outcome of in vitro fertilization and embryo transfer

    Institute of Scientific and Technical Information of China (English)

    赵静; 蔡霞; 胡泊; 史敏; 巴哈古丽·阿不拉; 杜红; 玛依热·吐尔逊

    2006-01-01

    目的:探讨体外受精-胚胎移植(in vitro fertilization and embryo transfer,IVF-ET)过程中基础卵泡刺激素(FSH)水平及获卵数对IVF-ET结局的影响.方法:对180例不孕患者共180个IVF-ET周期的资料进行回顾性分析,比较不同FSH水平和不同获卵数患者年龄、受精率、卵裂率、优质胚胎率和临床妊娠率等指标的差异.结果:不同FSH水平患者年龄、促性腺激素(Gn)剂量、Gn促排时间、移植胚胎数、卵裂率、妊娠率经统计学检验,差异均无统计学意义(P>0.05).FSH>8.78 IU/L者获卵数明显低于FSH<3.85 IU/L和FSH 3.85~8.78 IU/L组(P<0.05),而受精率和优质胚胎率却明显高于其它2组(P<0.05).随年龄增长、FSH水平增加获卵数逐渐减少,差异有统计学意义(P<0.05);不同获卵数者的受精率、卵裂率、优质胚胎率、临床妊娠率及中重度卵巢过度刺激综合征(OHSS)发生率,差异均有统计学意义(P<0.05),获卵数≤5个受精率最高,获卵数>15个受精率最低;获卵数6~15个卵裂率、优质胚胎率最高;随着获卵数的增加,临床妊娠率和中重度OHSS发生率也逐渐增高.获卵数和FSH水平呈显著性负相关(r=-0.246,P <0.01);经多因素Logistic回归分析,年龄、获卵数、移植胚胎数与IVF-ET妊娠结局具有相关性(P<0.05).结论:基础FSH水平不能预测IVF-ET的临床结局,而获卵数可以预测IVF-ET临床结局.

  1. Effect of two activation treatments and age of blastomere karyoplasts on in vitro development of bovine nuclear transfer embryos

    DEFF Research Database (Denmark)

    Booth, P J; Holm, P; Vajta, G;

    2001-01-01

    , or 5 in vitro produced donor embryos, were examined in order to define an optimal nuclear transfer protocol. The two activation protocols comprised calcium ionophore followed by either CHX or DMAP. Parthenogenetic blastocyst yields were greater (P

  2. Imprinting diseases and IVF: Danish National IVF cohort study

    DEFF Research Database (Denmark)

    Lidegaard, Ojvind; Pinborg, Anja; Andersen, Anders Nyboe

    2005-01-01

    The aim of this study was to compare the frequency of imprinting diseases in children born after IVF with the incidence in naturally conceived children.......The aim of this study was to compare the frequency of imprinting diseases in children born after IVF with the incidence in naturally conceived children....

  3. Clinical and logistical aspects of in vitro fertilization treatment: An analysis of a transport IVF programme

    OpenAIRE

    Roest, Jan

    1997-01-01

    textabstractIn vitro fertilization and cmblYo transfer (IVFIET), the culture of aspirated aGcytcs and spermatozoa in the JaboratOly follO\\\\ved by transccrvicai embryo transfer, ,vas originally used as a treatment for infertility resulting from impaired fUllction of the fallopian tubes. A few years after the introduction of IVF/ET the indications for treatment included infertility caused by endometriosis, male factors, ovulation disorders and unexplained infertility (1). Although IVF has now b...

  4. 微卵巢刺激Mini-IVF、玻璃化冷冻及冷冻胚胎移植%Minimal ovarian stimulation(mini-IVF),vitrification and frozen embryo transfer

    Institute of Scientific and Technical Information of China (English)

    张进

    2010-01-01

    @@ 为提供更自然的体外受精(IVF)治疗,Mini-IVF方案不断改进.在这非寻常的方案中,使用克罗米酚(雌激素拮抗剂)提高垂体卵泡刺激素(FSH)的分泌来刺激卵巢.克罗米酚持续使用超过5 d可以抑制黄体生成素(LH)峰[1].

  5. Pertinence of apoptosis markers for the improvement of in vitro fertilization (IVF).

    Science.gov (United States)

    Haouzi, D; Hamamah, S

    2009-01-01

    In assisted reproductive technology (ART), the pregnancy and birth rates following in vitro fertilization (IVF) attempts are still low. Recently, apoptotic markers have been suggested as new criteria for oocyte and embryo quality selection. Many studies have provided evidence that poor oocyte and embryo quality can be associated with apoptosis. The aim of this review is to summarize our current knowledge on the apoptotic process in oocytes and embryos, and focus on the possibility for using apoptotic markers as a reliable and predictive marker to select competent oocytes and embryos during IVF. Moreover, it is currently accepted that IVF failures, linked to poor embryo quality, are, in part, associated with suboptimal in vitro culture conditions. Here, we also review the current state of knowledge concerning how the genetic control of apoptosis during folliculogenesis and pre-implantation embryonic development is affected by in vitro culture conditions during IVF. In the future, identification of apoptotic markers in ART for oocyte and embryo selection should result in the development of new agonistic or antagonistic molecules of apoptosis by medicinal chemistry.

  6. Hepes na produção de embriões bovinos in vitro Hepes on in vitro production of bovine embryos

    Directory of Open Access Journals (Sweden)

    Marcelo Marcos Montagner

    2000-06-01

    of pH changes in maturation and embryo development media, buffered with different HEPES concentrations. Initially, the effect of different concentrations of HEPES (0, 12.5 and 25.0mM on the variation of pH in the maturation (modified TCM-199 and embryonic development (modified KSOM media was evaluated at room temperature (25ºC and in an atmosphere of 5% CO2 in air at 39ºC. In another experiment, the effect of HEPES on in vitro oocyte maturation was determined. Oocytes were maturated in TCM-199 modified either with 25.0mM of HEPES (HEPES group; n = 137 or without HEPES (control group; n = 142, performing 7 replicates and evaluating the rate of blastocyst. In this study, the medium used for fertilization was Fert-TALP while for embryo development was KSOM with 10% of fetal bovine serum with monolayer of oviduct epithelial cells. A third experiment was designed to determine the effect of HEPES on embryo development. The zygotes were divided in two groups and co-incubated with oviduct epithelial cells in modified KSOM with 10% of fetal bovine serum without HEPES (n = 95 or with 25.0mM of HEPES (n = 92. For this experiment, it was used embryos with two or more cells and the embryo development was considered from cleavage to expanded blastocyst (Bx, 7 and 9 days after insemination. The oocytes and embryos were incubated at temperature of 39ºC, an atmosphere containing 5% CO2 in air and saturated humidity. The media with 25.0mM of HEPES were more efficient in minimizing the range of pH than those with 12.5mM or without HEPES. To determine the effect of HEPES during in vitro oocyte maturation, the percentage of Bl considered either the total number of oocytes or the total number of cleavages was higher in the HEPES group (21.9% or 42.9%, respectively than those obtained in the control group (10.56% or 16.67%, respectively. When HEPES was added to embryo culture medium, the percentage of Bx (45.65% was higher than that obtained in medium without HEPES (11.58%; p<0.01. The

  7. Inheritance of resistance of bovine preimplantation embryos to heat shock: relative importance of the maternal versus paternal contribution.

    Science.gov (United States)

    Block, J; Chase, C C; Hansen, P J

    2002-09-01

    Brahman preimplantation embryos are less affected by exposure to heat shock than Holstein embryos. Two experiments were conducted to test whether the ability of Brahman embryos to resist the deleterious effects of heat shock was a result of the genetic and cellular contributions from the oocyte, spermatozoa, or a combination of both. In the first experiment, Brahman and Holstein oocytes were collected from slaughterhouse ovaries and fertilized with spermatozoa from an Angus bull. A different bull was used for each replicate to eliminate bull effects. On day 4 after fertilization, embryos >or= 9 cells were collected and randomly assigned to control (38.5 degrees C) or heat shock (41 degrees C for 6 hr) treatments. The proportion of embryos developing to the blastocyst (BL) and advanced blastocyst (ABL; expanded and hatched) stages was recorded on day 8. Heat shock reduced the number of embryos produced from Holstein oocytes that developed to BL (P Brahman oocytes (BL = 42.1 +/- 4.8% vs. 55.6 +/- 4.8% for 38.5 and 41 degrees C, respectively; ABL = 17.6 +/- 4.2% vs. 32.4 +/- 4.2%). In the second experiment, oocytes from Holstein cows were fertilized with semen from bulls of either Brahman or Angus breeds. Heat shock of embryos >or= 9 cells reduced development to BL (P Brahman (BL = 54.3 +/- 7.7% vs. 23.4 +/- 7.7%; ABL = 43. +/- 7.4% vs. 7.9 +/- 7.4%, for 38.5 and 41 degrees C, respectively) and Angus bulls (BL = 57.9 +/- 7.7% vs. 31.0 +/- 7.7%; ABL = 33.6 +/- 7.4% vs. 18.4 +/- 7.4%, for 38.5 and 41 degrees C, respectively). There were no breed x temperature interactions. Results suggest that the oocyte plays a more significant role in the resistance of Brahman embryos to the deleterious effects of heat shock than the spermatozoa. PMID:12211058

  8. The correlation analysis of AMH and Inhibin B levels in controlled ovarian stimulation with pregnancy outcome in vitro fertilization-embryo transfer%控制性促排卵中AMH和Inhibin B水平与IVF-ET结局的相关性分析

    Institute of Scientific and Technical Information of China (English)

    翟燕红; 李莹; 杨晓葵

    2015-01-01

    目的 探讨控制性促排卵过程中抗苗勒氏管激素(anti-müllerian hormone,AMH)和抑制素B (inhibin B,INHB)水平与体外受精(in vitro fertilization,IVF)/卵母细胞浆内单精子注射(intracytoplasmic sperm injection,ICSI)—胚胎移植(embryo transfer,ET)妊娠结局的相关性.方法 选择2012年1月至201 3年12月因输卵管因素和/或男方因素在我院生殖中心首次接IVF/ICSI-ET助孕患者为研究对象,共92个促排卵周期,检测促排卵周期中血清及取卵日卵泡液AMH和INHB水平,根据妊娠结局分为妊娠组和非妊娠组,比较两组患者的一般特点、内分泌激素水平、AMH和INHB水平、获卵数、受精率等促排卵相关参数,并进行相关性分析.结果 在妊娠组和非妊娠组中,患者的一般情况、激素水平、血清及卵泡液AMH和INHB水平、获卵数、受精率等促排卵相关参数均无统计学差异(P>0.05);血清AMH和INHB水平及卵泡液AMH水平均与窦卵泡数、hCG日雌二醇水平、获卵数存在显著正相关(P<0.001).结论 AMH和INHB水平可以反映促排卵周期中卵巢反应性,与IVF/ICSI-ET妊娠结局无直接相关性.

  9. Use of versapoint to refashion the cervical canal to overcome unusually difficult embryo transfers and improve in-vitro fertilization-embryo transfer outcome: A case series

    Directory of Open Access Journals (Sweden)

    Nalini Mahajan

    2011-01-01

    Full Text Available Background : Smooth atraumatic embryo transfer is paramount for the success of in-vitro fertilization (IVF. In difficult cases, cervical canal manipulation may be required. Aim : To see if surgical correction of the cervical canal or cervical canal refashioning could improve ease of embryo transfer. Setting : Private infertility and IVF hospital. Design : Prospective study. Materials and Methods : Patients: 11 women with failed 1-3 IVF cycles with history of extremely difficult embryo transfers (ETs despite undergoing cervical dilatation in the cycle prior to IVF. Interventions : Operative hysteroscopy using Versapoint for refashioning of the cervical canal. Main Outcome Measures : Ease of ET in the subsequent IVF cycle. Secondary outcome measure was to assess reproductive outcome. Results : Easy and atraumatic ET in the IVF cycle after procedure in 100% patients. PR was 46.5%. Conclusions : Use of Versapoint for refashioning the cervical canal can improve the quality of ET and PR.

  10. The role of RNA polymerase I transcription and embryonic genome activation in nucleolar development in bovine preimplantation embryos

    DEFF Research Database (Denmark)

    Østrup, Olga; Strejcek, F.; Petrovicova, I.;

    2008-01-01

    The aim of the present study was to investigate the role of RNA polymerase I (RPI) transcription in nucleolar development during major transcriptional activation (MTA) in cattle. Late eight-cell embryos were cultured in the absence (control group) or presence of actinomycin D (AD) (RPI inhibition...

  11. Factors affecting survival rates of in vitro produced bovine embryos after vitrification and direct in-straw rehydration

    DEFF Research Database (Denmark)

    Vajta, Gábor; Holm, Peter; Greve, Torben;

    1996-01-01

    The aim of this work was to investigate the possibilities of simplification, and to outline the limits of application, of a vitrification method for cow embryos. Morulae and blastocysts were produced by in vitro fertilization of slaughterhouse-derived, in vitro matured oocytes with frozen-thawed ...

  12. What is the optimal means of preparing the endometrium in frozenthawed embryo transfer cycles? A systematic review and meta-analysis

    NARCIS (Netherlands)

    Groenewoud, Eva R.; Cantineau, Astrid E. P.; Kollen, Boudewijn J.; Macklon, Nick S.; Cohlen, Ben J.

    2013-01-01

    Frozenthawed embryo transfer (FET) enables surplus embryos derived from IVF or IVF-ICSI treatment to be stored and transferred at a later date. In recent years the number of FET cycles performed has increased due to transferring fewer embryos per transfer and improved laboratory techniques. Currentl

  13. Comparison of Clinical Efficacy of in vitro Fertilization-embryo Transfer Versus two Different Protocols after Super-long Down-regulation%IVF-ET中超长方案降调节后两种促排卵方案临床疗效比较

    Institute of Scientific and Technical Information of China (English)

    阳翎; 汤倩倩; 高士友

    2015-01-01

    【目的】探讨在体外受精‐胚胎移植(IVF‐ET )治疗中采用超常方案降调节后分别使用尿源性卵泡刺激素(u‐FSH)和基因重组FSH(r‐FSH)联合尿促性腺激素(HMG)促排后的临床结局。【方法】回顾性分析2013年1~12本院生殖中心559例接受IVF或卵胞浆单精子注射(ICSI)助孕治疗的临床结局。【结果】两组患者促性腺激素(gonadotropin ,Gn)天数、Gn使用剂量,获卵数、受精率、卵裂率、妊娠率、可利用胚胎数、种植率、流产率、异位妊娠率、卵巢过度刺激综合征(OHSS)发生率均无显著差异( P >0.05)。【结论】IVF‐ET中超长方案降调节后u‐FSH联合HMG和r‐FSH联合 HMG促排卵的效果相当。%[Objective] To compare the clinical efficacy of in vitro fertilization‐embryo transfer (IVF‐ET ) versus two different kinds of follicle‐stimulating hormone (FSH) stimulation protocols for controlled hyper‐stimulation after super‐long down‐regulation .[Methods]A total of 559 IVF/intracytoplasmic sperm injection (ICSI)‐ET cycles in in‐fertile patients at our reproductive center from January to November 2013 were analyzed retrospectively .According to the ovarian stimulation protocolr‐FSH+75IUHMG ,they were divided into two groups of gonal‐F ( n =294) and urofollitropin group ( n=265) .The clinical outcomes were compared between two groups .[Results]The total dos‐age of gonadotropin ,duration of gonadotropin stimulation ,estradiol level on HCG day ,number of retrieved oocytes , fertilization rate ,number of tranferable embryos ,clinical pregnancy rate ,implantation rate ,abortion rate ,rate of ec‐topic pregnancy and incidence of ovarian hyperstimulation syndrome were not significantly different between two groups ( P>0 .05) .[Conclusion]A combination of u‐FSH or r‐FSH and HMG have the same effectiveness in ovari‐an stimulation during IVF‐ET cycles after super‐long down‐regulation .

  14. Evaluation of syphilis serostatus on the safety of IVF treatment.

    Science.gov (United States)

    Lin, Shengli; Li, Rong; Huang, Shuo; Zhao, Lianming; Li, Ming; Li, Junsheng; Zhu, Jinliang; Zheng, Xiaoying; Huang, Jin; Liu, Ping; Qiao, Jie

    2014-12-01

    An increasing number of infertile syphilis-infected individuals have turned to assisted reproductive technology; however, the safety of syphilis carrier serostatus on IVF and embryo transfer outcomes has not been evaluated. Data from 482 patients who delivered singletons were analysed. In the retrospective study, the rate of IVF and intracytoplasmic sperm injection fertilization was 79.50% ± 17.57%/78.72% ± 16.66% in the Treponema pallidum particle agglutination assay negative (TPPA-negative) and rapid plasma reagin negative (RPR-negative) group, 76.12% ± 22.99%/74.05% ± 20.31% in the TPPA-positive and RPR-negative group, and 75.66% ± 21.72%/70.90% ± 16.11% in the TPPA-positive and RPR-positive group. The clinical pregnancy rate was 39.79% in the TPPA-negative and RPR-negative group, 46.30% in the TPPA-positive and RPR-negative group, and 36.59% in the TPPA-positive and RPR-positive group. No significant differences were found between the groups. The neonatal gestational age and mean birth weight were not significantly different between the TPPA-negative and TPPA-positive groups. Multiple linear regression analysis also showed no association between TPPA serostatus and newborn birth weight and gestational age. The present retrospective study showed that TPPA and RPR serostatus did not affect the outcomes of IVF and embryo transfer. Syphilis-infected individuals can undergo IVF and embryo transfer cycles after penicillin treatment. PMID:25444510

  15. Evaluation of bovine (Bos indicus) ovarian potential for in vitro embryo production in the Adamawa plateau (Cameroon)

    OpenAIRE

    Kouamo, J.; Dawaye, S.M.; Zoli, A. P.; Bah, G. S.

    2014-01-01

    An abattoir study was conducted to evaluate the ovarian potential of 201 local zebu cattle from Ngaoundere, Adamawa region (Cameroon) for in vitro embryo production (IVEP). The ovaries were excised, submerged in normal saline solution (0.9%) and transported to the laboratory for a detailed evaluation. Follicles on each ovary were counted, their diameters (Φ) measured and were grouped into 3 categories: small (Φ 8 mm). Each ovary was then sliced in...

  16. Producción in vitro de embriones bovinos: suplementación de los medios de cultivo con suero In vitro production of bovine embryos: serum supplementation to the culture media

    Directory of Open Access Journals (Sweden)

    N Mucci

    2006-01-01

    Full Text Available Las técnicas para producir embriones bovinos, en estadios de preimplantación mediante la maduración de ovocitos y su posterior fertilización in vitro, ofrece la posibilidad de obtener embriones a bajo costo para ser utilizados con fines de estudio (desarrollo embrionario temprano, transgénesis, clonación o con propósitos comerciales. Las condiciones de cultivo in vitro pueden influenciar significativamente el desarrollo embrionario, determinando cambios responsables de su menor calidad, comparados con los embriones producidos in vivo . En particular, la adición de suero a los medios de cultivo altera tanto la morfología embrionaria como su calidad, y su eliminación posibilitaría producir embriones de buena calidad para ser criopreservados y transferidos. Los objetivos de esta revisión son describir los aspectos generales del cultivo in vitro de embriones y resumir algunas hipótesis referidas a los mecanismos por los cuales el suero podría alterar el desarrollo y la calidad embrionaria.Techniques for producing bovine preimplantation embryos by in vitro maturation and fertilization, offers the potential for a large number of embryos at low cost to be used for basic scientific research (embryo development, transgenesis, cloning or for commercial purposes. Embryo culture conditions can influence in vitro embryo development significantly, provoking deviations responsible for low quality compared with in vivo counterparts. In particular, serum supplementation alters both embryo morphology and quality and eliminating serum from the media could be beneficial to produce good quality embryos for cryopreservation and embryo transfer. The objectives of this review are to describe the general aspects of in vitro embryo culture and summerize some hypotheses about the way serum could alter embryo development and quality.

  17. Bivariate analysis of basal serum anti-Mullerian hormone measurements and human blastocyst development after IVF

    LENUS (Irish Health Repository)

    Sills, E Scott

    2011-12-02

    Abstract Background To report on relationships among baseline serum anti-Müllerian hormone (AMH) measurements, blastocyst development and other selected embryology parameters observed in non-donor oocyte IVF cycles. Methods Pre-treatment AMH was measured in patients undergoing IVF (n = 79) and retrospectively correlated to in vitro embryo development noted during culture. Results Mean (+\\/- SD) age for study patients in this study group was 36.3 ± 4.0 (range = 28-45) yrs, and mean (+\\/- SD) terminal serum estradiol during IVF was 5929 +\\/- 4056 pmol\\/l. A moderate positive correlation (0.49; 95% CI 0.31 to 0.65) was noted between basal serum AMH and number of MII oocytes retrieved. Similarly, a moderate positive correlation (0.44) was observed between serum AMH and number of early cleavage-stage embryos (95% CI 0.24 to 0.61), suggesting a relationship between serum AMH and embryo development in IVF. Of note, serum AMH levels at baseline were significantly different for patients who did and did not undergo blastocyst transfer (15.6 vs. 10.9 pmol\\/l; p = 0.029). Conclusions While serum AMH has found increasing application as a predictor of ovarian reserve for patients prior to IVF, its roles to estimate in vitro embryo morphology and potential to advance to blastocyst stage have not been extensively investigated. These data suggest that baseline serum AMH determinations can help forecast blastocyst developmental during IVF. Serum AMH measured before treatment may assist patients, clinicians and embryologists as scheduling of embryo transfer is outlined. Additional studies are needed to confirm these correlations and to better define the role of baseline serum AMH level in the prediction of blastocyst formation.

  18. Altered cleavage patterns in human tripronuclear embryos and their association to fertilization method

    DEFF Research Database (Denmark)

    Joergensen, Mette Warming; Agerholm, Inge; Hindkjaer, Johnny;

    2014-01-01

    PURPOSE: To analyze the cleavage patterns in dipronuclear (2PN) and tripronuclear (3PN) embryos in relation to fertilization method. METHOD: Time-lapse analysis. RESULTS: Compared to 2PN, more 3PN IVF embryos displayed early cleavage into 3 cells (p ... stage (p time frame. However, timing of the cell divisions within the cleavage cycles differed between the two groups. In contrast.......001). More 3PN IVF than ICSI embryos displayed early cleavage into 3 cells (p time demonstrates differences in the cleavage pattern between 3PN IVF and ICSI embryos....

  19. Milder ovarian stimulation for in-vitro fertilization reduces aneuploidy in the human preimplantation embryo: A randomized controlled trial

    NARCIS (Netherlands)

    E.B. Baart (Esther); E. Martini (Elena); M.J.C. Eijkemans (René); D. van Opstal (Diane); N.G.M. Beckers (Nicole); A. Verhoeff (Arie); N.S. Macklon (Nick); B.C.J.M. Fauser (Bart)

    2007-01-01

    textabstractBACKGROUND: To test whether ovarian stimulation for in-vitro fertilization (IVF) affects oocyte quality and thus chromosome segregation behaviour during meiosis and early embryo development, preimplantation genetic screening of embryos was employed in a prospective, randomized controlled

  20. 胚胎移植前宫腔内灌注hCG在IVF-ET中的应用及hCG提高种植率和妊娠率的有关机制%Application of the Established Method of Intrauterine Infusion of hCG before Embryo Transfer in IVF-ET and Mechanisms of hCG to Improve the Implantation and Pregnancy Rates

    Institute of Scientific and Technical Information of China (English)

    王会妍; 张帅; 张云山

    2012-01-01

    人绒毛膜促性腺激素(human chorionic gonadotropin,hCG)是由胎盘合体滋养细胞合成的一种糖蛋白激素,具有维持黄体寿命、刺激雌孕激素合成、抑制植物血凝素对淋巴细胞的增殖作用等功能.最近研究表明,胚胎移植前宫腔内灌注hCG可显著提高移植后的种植率和妊娠率,但其具体作用机制尚未完全弄清,可能与宫腔内灌注一定浓度hCG后,其能以旁分泌形式作用于子宫内膜上皮细胞和血管内皮细胞等的表面受体而发挥作用,进而促使母-胎界面血管形成、增强母-胎间的免疫耐受有关;还可能与延长子宫内膜的移植窗,使子宫内膜容受性增强有关.%Human chorionic gonadotropin (hCG), a glycoprotein which is synthesized by the placenta syncytiotrophoblasts has multiple functions, such as maintaining the life of corpus luteum, stimulating the synthesis of estrogen and progesterone, inhibiting the proliferation reaction of lymphocytes responding to phytohemagglutinin and so on. A recent study has found that the application of intrauterine infusion of hCG before embryo transfer can significantly improve the implantation and pregnancy rates in IVF-ET, however, its' exact regulating mechanism is not fully understood by researchers, it may be explained by the previous proved value of hCG, for example, when it was infused into the uterine cavity up to a certain concentration, it can play a paracrine role on promoting immunologic tolerance-and angiogenesis at the maternal-fetal interface via its interaction with endometrial and endothelial hCG receptors; in addition, it may in part be explained by increasing the duration of the implantation window of endometrial as well as enhancing the receptivity of uterine.

  1. What Is the Value of Three-Parent IVF?

    Science.gov (United States)

    Rulli, Tina

    2016-07-01

    In February 2016, the Institute of Medicine released a report, commissioned by the United States Food and Drug Administration, on the ethical and social-policy implications of so-called three-parent in vitro fertilization. The IOM endorses commencement of clinical trials on three-parent IVF, subject to some initial limitations. Also called mitochondrial replacement or transfer, three-parent IVF is an intervention comprising two distinct procedures in which the genetic materials of three people-the DNA of the father and mother and the mitochondrial DNA of an egg donor-can be used to create a child. Three-parent IVF would enable a woman with mitochondrial disease to have a genetically related child without transmitting the disease to the child. The possibility for three-parent children has prompted criticism from many corners. Critics have pointed to ethical issues including safety concerns and risks to children, genetic and germline engineering concerns, the potential exploitation of the third-parent egg donor, donor anonymity and privacy, and objections to creating babies with three parents, which undermines natural and traditional conceptions of procreation. Additionally, developing the technology would involve experimenting on, manipulating, and disposing of embryos. Although the IOM report considers the ethical concerns about the value of the three-parent IVF technology, the IOM failed to give due attention to an important objection to the development of this technology: three-parent IVF lacks the social value necessary to make investment of public resources in it ethical. Unlike the other concerns, this objection is not based on conservativism about new reproductive technologies or default favoritism of the status quo. I argue that the technology does not meet a plausible social value standard to render public research investment into its development ethical. Proponents of three-parent IVF make inaccurate and exaggerated claims that it will eradicate

  2. What Is the Value of Three-Parent IVF?

    Science.gov (United States)

    Rulli, Tina

    2016-07-01

    In February 2016, the Institute of Medicine released a report, commissioned by the United States Food and Drug Administration, on the ethical and social-policy implications of so-called three-parent in vitro fertilization. The IOM endorses commencement of clinical trials on three-parent IVF, subject to some initial limitations. Also called mitochondrial replacement or transfer, three-parent IVF is an intervention comprising two distinct procedures in which the genetic materials of three people-the DNA of the father and mother and the mitochondrial DNA of an egg donor-can be used to create a child. Three-parent IVF would enable a woman with mitochondrial disease to have a genetically related child without transmitting the disease to the child. The possibility for three-parent children has prompted criticism from many corners. Critics have pointed to ethical issues including safety concerns and risks to children, genetic and germline engineering concerns, the potential exploitation of the third-parent egg donor, donor anonymity and privacy, and objections to creating babies with three parents, which undermines natural and traditional conceptions of procreation. Additionally, developing the technology would involve experimenting on, manipulating, and disposing of embryos. Although the IOM report considers the ethical concerns about the value of the three-parent IVF technology, the IOM failed to give due attention to an important objection to the development of this technology: three-parent IVF lacks the social value necessary to make investment of public resources in it ethical. Unlike the other concerns, this objection is not based on conservativism about new reproductive technologies or default favoritism of the status quo. I argue that the technology does not meet a plausible social value standard to render public research investment into its development ethical. Proponents of three-parent IVF make inaccurate and exaggerated claims that it will eradicate

  3. Efecto del ácido linoleico conjugado sobre la proporción de sexos y calidad de embriones bovinos producidos in vitro Effect of conjugated linoleic acid on sex ratio and quality of in vitro produced bovine embryos

    Directory of Open Access Journals (Sweden)

    NA Gómez

    2013-01-01

    Full Text Available El objetivo de este estudio fue evaluar el efecto de la suplementación del medio de cultivo con ácido linoleico conjugado (CLA sobre el clivaje, producción, proporción de sexos y calidad embrionaria en embriones bovinos producidos in vitro al día 7 de cultivo. Se fertilizaron 308 CCO suplementados en cultivo con 100 µM del isómero de CLA Cis-9 Trans-11 y Cis-10-Trans-12 y 257 CCO en el grupo control; la producción de embriones fue 25,32% vs 35,40% respectivamente con diferencia significativa (P The aim of this study was to evaluate the effect of culture medium supplementation with conjugated linoleic acid (CLA on embryo cleavage, embryo production, sex ratio and embry o quality in in vitro produced bovine embryos at day 7 of culture. 308 COCs were used for the group supplemented with 100 µM of the CLA isomer Cis-9 trans-11 and Cis-10-Trans-12 and 257 COCs for the untreated control group; the embryo production was 25.32% vs 35.40%, respectively, with significant difference between them (P < 0.05. The embryos were classified according to the IETS in Mo, Bt, Bl and Bx stages for morphological and molecular analysis. PCR was used for sex determination; embryo quality was assessed as grade 1 (excellent or good and Grade 2 (regular. The results showed no significant difference in the proportion of embryos male:female for any of the stages in the CLA supplemented group achieving the expected natural ratio (50:50, while the control maintained a greater number of males. The CLA improved quality in Bl and Bt stages for both females and males (P < 0.05 having a greater number of grade 1 embryos in supplemented group, while control embryos were more in grade 2. In conclusion, CLA adversely affects the production of bovine embryos in vitro, but the sex ratio equals the natural one in all stages and improves embryo quality in some stages of early development.

  4. Vitrificación como técnica de crioconservación de embriones bovinos Vitrification as a technique of bovine embryo cryopreservation

    Directory of Open Access Journals (Sweden)

    M CELESTINOS

    2002-01-01

    less expensive freezing procedures. One of these is vitrification, a process of solidification that uses a highly concentrated solution which does not crystallize during freezing; its viscosity increases with the descent of the temperature until the formation of an amorphous solid state similar to glass. For this reason, exposure and freezing rates should be quick enough to avoid toxicity and the formation of intracellular ice, which can cause embryonic damage. In order for the embryos to support the osmotic shock, they should be equilibriated with a less concentrated crioprotectant solution before being exposed to the vitrificant solution for their later freezing. Several vitrification procedures have been published about embryo preservation, using different crioprotectants, concentration, volume, addition method, temperatures, exposition time, freezing rate, thawing procedure and dilution, to maintain the function, normal structure and viability of the embryo. These techniques have also been experimented with in vitro and in vivo produced embryos, at different developmental stages. This paper aims to review the present bovine embryo cryopreservation methods, particulary vitrification, as well as to mention the latest procedures and progress so that new researchers may have an updated literature review to start their works.

  5. 不同分型的多囊卵巢综合征患者 IVF-ET 后的妊娠结局%Pregnancy outcomes of different subtypes of polycystic ovary syndrome patients after in vitro fertilization and embryo transfer

    Institute of Scientific and Technical Information of China (English)

    刘大艳; 朱文杰; 付志红; 李雪梅

    2015-01-01

    Objective To investigate the pregnancy outcomes of different subtypes of polycystic ovary syndrome ( PCOS) patients after in vitro fertilization and embryo transfer ( IVF-ET) .Methods Retrospective study was conducted among 160 pregnant women with PCOS and 350 infertile women with tubal factors who undergoing IVF-ET.They were divided into four groups:28 cases in obese-PCOS group ( group A) , 132 cases in non-obese PCOS group ( group B) , 41 cases in obese tubal factor group ( group C) , and 309 cases in non-obese tubal factor group (group D) with obesity defined by body mass index (BMI) over 24kg/m2.Clinical characteristics, IVF parameters, pregnancy and perinatal outcomes were analyzed in each group.Results There were significant differences among four groups in age, BMI, proportion of primary infertility, infertility duration and basal FSH level ( t=12.603, P=0.031; t=21.619, P=0.000;χ2 =78.681, P=0.000;t=7.838, P=0.045;t=16.264, P=0.020).The E2 level in hCG day was highest in group B, and the number of retrieved oocytes was also the most, followed by group A, both of which were significantly more than tubal factor groups ( group C and D) ( t=28.581, P=0.000;t=25.492, P=0.000).The fertilization rate, the number of good embryos and the implantation rate of PCOS groups (group A and B) were lower than those of the tubal groups (group C and D), and the differences were significant (t=21.642, P=0.000;t=9.863, P=0.035;χ2 =27.711, P=0.000).The morbidity of GDM and the incidence of large for gestational age infant in obese groups (group A and C) were obviously higher than those in non-obese groups (group B and D) (χ2 =14.698, P=0.002;χ2 =25.003, P=0.000) .The incidence of small for gestational age infant was higher in the PCOS groups ( group A and B) than in the tubal factor groups (group C and D), and the difference was significant (χ2 =8.086, P=0.044).Conclusion There are some differences in IVF parameters, pregnancy and perinatal outcomes between obese and non

  6. Pregnancy outcomes of different subtypes of polycystic ovary syndrome patients after in vitro fertilization and embryo transfer%不同分型的多囊卵巢综合征患者 IVF-ET 后的妊娠结局

    Institute of Scientific and Technical Information of China (English)

    刘大艳; 朱文杰; 付志红; 李雪梅

    2015-01-01

    Objective To investigate the pregnancy outcomes of different subtypes of polycystic ovary syndrome ( PCOS) patients after in vitro fertilization and embryo transfer ( IVF-ET) .Methods Retrospective study was conducted among 160 pregnant women with PCOS and 350 infertile women with tubal factors who undergoing IVF-ET.They were divided into four groups:28 cases in obese-PCOS group ( group A) , 132 cases in non-obese PCOS group ( group B) , 41 cases in obese tubal factor group ( group C) , and 309 cases in non-obese tubal factor group (group D) with obesity defined by body mass index (BMI) over 24kg/m2.Clinical characteristics, IVF parameters, pregnancy and perinatal outcomes were analyzed in each group.Results There were significant differences among four groups in age, BMI, proportion of primary infertility, infertility duration and basal FSH level ( t=12.603, P=0.031; t=21.619, P=0.000;χ2 =78.681, P=0.000;t=7.838, P=0.045;t=16.264, P=0.020).The E2 level in hCG day was highest in group B, and the number of retrieved oocytes was also the most, followed by group A, both of which were significantly more than tubal factor groups ( group C and D) ( t=28.581, P=0.000;t=25.492, P=0.000).The fertilization rate, the number of good embryos and the implantation rate of PCOS groups (group A and B) were lower than those of the tubal groups (group C and D), and the differences were significant (t=21.642, P=0.000;t=9.863, P=0.035;χ2 =27.711, P=0.000).The morbidity of GDM and the incidence of large for gestational age infant in obese groups (group A and C) were obviously higher than those in non-obese groups (group B and D) (χ2 =14.698, P=0.002;χ2 =25.003, P=0.000) .The incidence of small for gestational age infant was higher in the PCOS groups ( group A and B) than in the tubal factor groups (group C and D), and the difference was significant (χ2 =8.086, P=0.044).Conclusion There are some differences in IVF parameters, pregnancy and perinatal outcomes between obese and non

  7. Elevated progesterone during ovarian stimulation for IVF

    DEFF Research Database (Denmark)

    Al-Azemi, M; Kyrou, D; Kolibianakis, E M;

    2012-01-01

    There is an ongoing debate regarding the impact of premature progesterone rise on the IVF outcome. The objective of this review is to assess evidence of poorer ongoing pregnancy rate in IVF cycles with elevated serum progesterone at the end of follicular phase in ovarian stimulation. It also...... of premature progesterone in stimulated IVF cycles. There is an ongoing debate regarding the impact of premature progesterone rise on the IVF outcome. The objective of this review is to assess evidence of poorer ongoing pregnancy rate in IVF cycles with elevated serum progesterone at the end of follicular...... should document the cause and origin of premature progesterone in stimulated IVF cycles....

  8. IVF for premature ovarian failure: first reported births using oocytes donated from a twin sister

    Directory of Open Access Journals (Sweden)

    Sills Eric

    2010-03-01

    Full Text Available Abstract Background Premature ovarian failure (POF remains a clinically challenging entity because in vitro fertilisation (IVF with donor oocytes is currently the only treatment known to be effective. Methods A 33 year-old nulligravid patient with a normal karyotype was diagnosed with POF; she had a history of failed fertility treatments and had an elevated serum FSH (42 mIU/ml. Oocytes donated by her dizygotic twin sister were used for IVF. The donor had already completed a successful pregnancy herself and subsequently produced a total of 10 oocytes after a combined FSH/LH superovulation regime. These eggs were fertilised with sperm from the recipient's husband via intracytoplasmic injection and two fresh embryos were transferred to the recipient on day three. Results A healthy twin pregnancy resulted from IVF; two boys were delivered by caesarean section at 39 weeks' gestation. Additionally, four embryos were cryopreserved for the recipient's future use. The sister-donor achieved another natural pregnancy six months after oocyte retrieval, resulting in a healthy singleton delivery. Conclusion POF is believed to affect approximately 1% of reproductive age females, and POF patients with a sister who can be an oocyte donor for IVF are rare. Most such IVF patients will conceive from treatment using oocytes from an anonymous oocyte donor. This is the first report of births following sister-donor oocyte IVF in Ireland. Indeed, while sister-donor IVF has been successfully undertaken by IVF units elsewhere, this is the only known case where oocyte donation involved twin sisters. As with all types of donor gamete therapy, pre-treatment counselling is important in the circumstance of sister oocyte donation.

  9. IVF for premature ovarian failure: first reported births using oocytes donated from a twin sister.

    LENUS (Irish Health Repository)

    Sills, Eric Scott

    2010-01-01

    BACKGROUND: Premature ovarian failure (POF) remains a clinically challenging entity because in vitro fertilisation (IVF) with donor oocytes is currently the only treatment known to be effective. METHODS: A 33 year-old nulligravid patient with a normal karyotype was diagnosed with POF; she had a history of failed fertility treatments and had an elevated serum FSH (42 mIU\\/ml). Oocytes donated by her dizygotic twin sister were used for IVF. The donor had already completed a successful pregnancy herself and subsequently produced a total of 10 oocytes after a combined FSH\\/LH superovulation regime. These eggs were fertilised with sperm from the recipient\\'s husband via intracytoplasmic injection and two fresh embryos were transferred to the recipient on day three. RESULTS: A healthy twin pregnancy resulted from IVF; two boys were delivered by caesarean section at 39 weeks\\' gestation. Additionally, four embryos were cryopreserved for the recipient\\'s future use. The sister-donor achieved another natural pregnancy six months after oocyte retrieval, resulting in a healthy singleton delivery. CONCLUSION: POF is believed to affect approximately 1% of reproductive age females, and POF patients with a sister who can be an oocyte donor for IVF are rare. Most such IVF patients will conceive from treatment using oocytes from an anonymous oocyte donor. This is the first report of births following sister-donor oocyte IVF in Ireland. Indeed, while sister-donor IVF has been successfully undertaken by IVF units elsewhere, this is the only known case where oocyte donation involved twin sisters. As with all types of donor gamete therapy, pre-treatment counselling is important in the circumstance of sister oocyte donation.

  10. [Bovine progressive degenerative myeloencephalopathy ("Weaver syndrome") in brown Swiss x Braunvieh cattle: reproductive occurrences, results of embryo transfer].

    Science.gov (United States)

    Tenhumberg, H; Trela, T; Matzke, P; Averdunk, G; Dirksen, G

    1994-12-01

    In the time between 1989 and 1991 seven Brown Swiss heifers, which had clinical signs of the Weaver syndrome were kept at the Bavarian Institute of Animal Breeding in Grub. This was in order to investigate this hereditary trait further. The number of animals carrying this genetic defect was increased by means of embryo transfer. Both cycle observations and ovary controls by means of rectal palpation resulted largely in physiological data and findings. All seven animals responded to superovulation treatment which was induced by sequential doses of p-FSH (32 mg) or of a single dose of 2,000 IU PMSG. The donors were flushed a total of 32 times without problem. On average 5.3 ova were recovered, 2.8 of which were viable and suitable for transfer. These are statistically only 50% of the normal value in a routine ET programme. Following the transfer of fresh and frozen embryos the pregnancy rate was 53%. There was only one abortion observed from 48 pregnancies.

  11. Reduced developmental competence of immature, in-vitro matured and postovulatory aged mouse oocytes following IVF and ICSI

    OpenAIRE

    Trounson Alan; Lacham-Kaplan Orly

    2008-01-01

    Abstract Background The present study highlights basic physiological differences associated with oocyte maturation and ageing. The study explores the fertilizing capacity and resistance to injury of mouse oocytes at different stages of maturation and ageing following IVF and ICSI. Also, the study examines the developmental competence of embryos obtained from these oocytes. The outcome of the study supports views that the mouse can be a model for human IVF suggesting that utilizing in-vitro ma...

  12. SGO1 maintains bovine meiotic and mitotic centromeric cohesions of sister chromatids and directly affects embryo development.

    Directory of Open Access Journals (Sweden)

    Feng-Xia Yin

    Full Text Available Shugoshin (SGO is a critical factor that enforces cohesion from segregation of paired sister chromatids during mitosis and meiosis. It has been studied mainly in invertebrates. Knowledge of SGO(s in a mammalian system has only been reported in the mouse and Hela cells. In this study, the functions of SGO1 in bovine oocytes during meiotic maturation, early embryonic development and somatic cell mitosis were investigated. The results showed that SGO1 was expressed from germinal vesicle (GV to the metaphase II stage. SGO1 accumulated on condensed and scattered chromosomes from pre-metaphase I to metaphase II. The over-expression of SGO1 did not interfere with the process of homologous chromosome separation, although once separated they were unable to move to the opposing spindle poles. This often resulted in the formation of oocytes with 60 replicated chromosomes. Depletion of SGO1 in GV oocytes affected chromosomal separation resulting in abnormal chromosome alignment at a significantly higher proportion than in control oocytes. Knockdown of SGO1 expression significantly decreased the embryonic developmental rate and quality. To further confirm the function(s of SGO1 during mitosis, bovine embryonic fibroblast cells were transfected with SGO1 siRNAs. SGO1 depletion induced the premature dissociation of chromosomal cohesion at the centromere and along the chromosome arm giving rise to abnormal appearing mitotic patterns. The results of this study infer that SGO1 is involved in the centromeric cohesion of sister chromatids and chromosomal movement towards the spindle poles. Depletion of SGO1 causes arrestment of cell division in meiosis and mitosis.

  13. The promising and potential role of IVF in cattle and beef industry

    Directory of Open Access Journals (Sweden)

    Debasmita Debnath

    2013-06-01

    Full Text Available Background and Objective : IVF is one of the Assisted Reproductive technology . The study was conducted to reveal the importance of IVF in cattle and beef industries . It was revealed that IVF has a dynamic advantage. In addition , it is not necessary to superovulate the cattles and on the other hand a cattle can be aspirated every 20 days. As cattle can be harvested at a very young age , it reduces the generation interval with desirable trait. This technology is tremendously used in reproductive management strategies for improving pregnancy with low fertility. Method : In this present work oocytes were collected and were fertilized with sperm under laboratory procedure following the transfer of fertilized egg to the uterus of surrogate mother. Oocytes were collected by punction of 2-4 nm follicles by aspiration method. Stimulated with hormone FSH-P follicular fluid and few cumulous cells. Further , oocytes were washed repeatedly in the washing media and then transferred in fertilization media and inseminated with sperm , which gives rise to the embryo. Result :The work was performed by taking a total of 46 oocytes ; out of which 19 were matured and among them 7 were developed in embryos. The result of the study witnessed that the success rate of such In-vitro- produced embryos is 15%. Conclusion : The In-vitro produced embryos play a major role in genetic selection scheme based on allelic variance of specific gene. The method might suggests that further research is required to improve the aspects of IVF.

  14. Live births achieved via IVF are increased by improvements in air quality and laboratory environment

    Science.gov (United States)

    Heitmann, Ryan J; Hill, Micah J; James, Aidita N; Schimmel, Tim; Segars, James H; Csokmay, John M; Cohen, Jacques; Payson, Mark D

    2016-01-01

    Infertility is a common disease, which causes many couples to seek treatment with assisted reproduction techniques. Many factors contribute to successful assisted reproduction technique outcomes. One important factor is laboratory environment and air quality. Our facility had the unique opportunity to compare consecutively used, but separate assisted reproduction technique laboratories, as a result of a required move. Environmental conditions were improved by strategic engineering designs. All other aspects of the IVF laboratory, including equipment, physicians, embryologists, nursing staff and protocols, were kept constant between facilities. Air quality testing showed improved air quality at the new IVF site. Embryo implantation (32.4% versus 24.3%; P < 0.01) and live birth (39.3% versus 31.8%, P < 0.05) were significantly increased in the new facility compared with the old facility. More patients met clinical criteria and underwent mandatory single embryo transfer on day 5 leading to both a reduction in multiple gestation pregnancies and increased numbers of vitrified embryos per patient with supernumerary embryos available. Improvements in IVF laboratory conditions and air quality had profound positive effects on laboratory measures and patient outcomes. This study further strengthens the importance of the laboratory environment and air quality in the success of an IVF programme. PMID:26194882

  15. Replication of somatic micronuclei in bovine enucleated oocytes

    Directory of Open Access Journals (Sweden)

    Canel Natalia

    2012-11-01

    Full Text Available Abstract Background Microcell-mediated chromosome transfer (MMCT was developed to introduce a low number of chromosomes into a host cell. We have designed a novel technique combining part of MMCT with somatic cell nuclear transfer, which consists of injecting a somatic micronucleus into an enucleated oocyte, and inducing its cellular machinery to replicate such micronucleus. It would allow the isolation and manipulation of a single or a low number of somatic chromosomes. Methods Micronuclei from adult bovine fibroblasts were produced by incubation in 0.05 μg/ml demecolcine for 46 h followed by 2 mg/ml mitomycin for 2 h. Cells were finally treated with 10 μg/ml cytochalasin B for 1 h. In vitro matured bovine oocytes were mechanically enucleated and intracytoplasmatically injected with one somatic micronucleus, which had been previously exposed [Micronucleus- injected (+] or not [Micronucleus- injected (−] to a transgene (50 ng/μl pCX-EGFP during 5 min. Enucleated oocytes [Enucleated (+] and parthenogenetic [Parthenogenetic (+] controls were injected into the cytoplasm with less than 10 pl of PVP containing 50 ng/μl pCX-EGFP. A non-injected parthenogenetic control [Parthenogenetic (−] was also included. Two hours after injection, oocytes and reconstituted embryos were activated by incubation in 5 μM ionomycin for 4 min + 1.9 mM 6-DMAP for 3 h. Cleavage stage and egfp expression were evaluated. DNA replication was confirmed by DAPI staining. On day 2, Micronucleus- injected (−, Parthenogenetic (− and in vitro fertilized (IVF embryos were karyotyped. Differences among treatments were determined by Fisher′s exact test (p≤0.05. Results All the experimental groups underwent the first cell divisions. Interestingly, a low number of Micronucleus-injected embryos showed egfp expression. DAPI staining confirmed replication of micronuclei in most of the evaluated embryos. Karyotype analysis revealed that all Micronucleus-injected embryos had

  16. Can IVF influence human evolution?

    Science.gov (United States)

    Hanevik, Hans Ivar; Hessen, Dag O; Sunde, Arne; Breivik, Jarle

    2016-07-01

    IVF, a procedure in which pharmacological and technological manipulation is used to promote pregnancy, offers help to infertile couples by circumventing selection at the most fundamental level. Fertility is clearly one of the key fitness-promoting drivers in all forms of sexually reproducing life, and fertilization and pregnancy are fundamental evolutionary processes that involve a range of pre- and post-zygotic screening mechanisms. Here, we discuss the various selection and screening factors involved in fertilization and pregnancy and assess IVF practices in light of these factors. We then focus on the possible consequences of these differences in selection pressures, mainly at the individual but also at the population level, to evaluate whether changes in the reproducing genotype can affect human evolution. The aim of the article is not to argue for or against IVF, but to address aspects of assisted reproduction in an evolutionary context. PMID:27094480

  17. Embryo apoptosis identification: Oocyte grade or cleavage stage?

    OpenAIRE

    Bakri, Noraina Mohd; Ibrahim, Siti Fatimah; Osman, Nurul Atikah; Hasan, Nurhaslina; Jaffar, Farah Hanan Fathihah; Rahman, Zulaiha Abdul; Osman, Khairul

    2015-01-01

    Apoptosis is a programed cell death that is vital for tissue homeostasis. However, embryo apoptosis had been known to be related to embryo fragmentation which should be avoided in in vitro fertilization (IVF). The purpose of this study was to evaluate the relationship of embryo apoptosis with the grade of immature oocytes and cleavage stage of in vitro produced (IVP) cattle embryos. This study consisted of 345 oocytes collected through ovary slicing. Immature oocytes were graded as A, B and C...

  18. Comparison of the major malformation rate of children conceived from cryopreserved embryos and fresh embryos

    Institute of Scientific and Technical Information of China (English)

    LI Hong-zhen; QIAO Jie; CHI Hong-bin; CHEN Xin-na; LIU Ping; MA Cai-hong

    2010-01-01

    Background Cryopreserved embryo transfer has become indispensable in reproductive technology. More and more children are conceived from frozen-thawed embryo transfer (FET). The risk of birth defects associated with frozen-thawed embryo transfer has been evaluated and conflict results are obtained. The aim of this study was to compare the rate of major malformations in children conceived from cryopreserved embryos with that of children from fresh embryos. Methods A retrospective analysis was performed on children conceived from frozen-thawed embryos and fresh embryos between January 2005 and December 2008 at the Reproduction Center of the Third Hospital, Peking University.The major malformation rates were compared between two groups for all children, as well as singletons or twins,separately. The frequencies of different subtypes of malformations classified according to different organ system were also compared.Results Thirty-four of 3125 children from cryopreserved embryos had a major malformation. The malformation rate was 1.09%, which was comparable to that for children after fresh embryos transfer (1.53%(55/3604), OR:0.71, 95% CI; 0.46-1.09). The malformation rate was also similar when the analysis was limited to children from cryopreserved embryos resulted from in vitro fertilization (IVF)(1.39%)and fresh IVF(1.3%). However, children from cryopreserved embryos resulted from intracytoplasmic sperm injections (ICSI) had much lower malformation rate than from fresh ICSI(0.63% vs.1.83%, OR: 0.34, 95% CI: 0.16-0.75). No difference was found in the incidence of major malformations in singletons from cryo ICSI (0.73%) and fresh ICS1(1.9%), or from cryo IVF(1.49%) and fresh IVF(1.67%). Similar malformation rate was found in multiples from cryo ICSI(0.52%) and fresh ICSI(1.76%), or cryo IVF(1.30%) and fresh IVF(0.90%). The distribution and risk of the subtype of malformations, such as cardiovascular, gastrointestinal, neural tube, urogenital, rnusculoskeletal and facial

  19. Embryo quality and impact of specific embryo characteristics on ongoing implantation in unselected embryos derived from modified natural cycle in vitro fertilization

    NARCIS (Netherlands)

    Pelinck, Marie-Jose; Hoek, Annemieke; Simons, Arnold H. M.; Heineman, Maas Jan; van Echten-Arends, Janny; Arts, Eus G. J. M.

    2010-01-01

    Objective: To study the implantation potential of unselected embryos derived from modified natural cycle IVF according to their morphological characteristics. Design: Cohort study. Setting: Academic department of reproductive medicine. Patient(S): A series of 449 single embryo transfers derived from

  20. First line fertility treatment strategies regarding IUI and IVF require clinical evidence.

    Science.gov (United States)

    Bahadur, G; Homburg, R; Muneer, A; Racich, P; Alangaden, T; Al-Habib, A; Okolo, S

    2016-06-01

    The advent of intracytoplasmic sperm injection (ICSI) has contributed to a significant growth in the delivery of assisted conception technique, such that IVF/ICSI procedures are now recommended over other interventions. Even the UK National Institute for Health Care Excellence (NICE) guidelines controversially recommends against intrauterine insemination (IUI) procedures in favour of IVF. We reflect on some of the clinical, economic, financial and ethical realities that have been used to selectively promote IVF over IUI, which is less intrusive and more patient friendly, obviates the need for embryo storage and has a global application. The evidence strongly favours IUI over IVF in selected couples and national funding strategies should include IUI treatment options. IUI, practised optimally as a first line treatment in up to six cycles, would also ease the pressures on public funds to allow the provision of up to three IVF cycles for couple who need it. Fertility clinics should also strive towards ISO15189 accreditation standards for basic semen diagnosis for male infertility used to triage ICSI treatment, to reduce the over-diagnosis of severe male factor infertility. Importantly, there is a need to develop global guidelines on inclusion policies for IVF/ICSI procedures. These suggestions are an ethically sound basis for constructing the provision of publicly funded fertility treatments. PMID:27076499

  1. Storage of bovine isolated follicles: a new alternative way to improve the recovery rate of viable embryos from ovarian follicles of slaughtered cows.

    Science.gov (United States)

    Pavlok, A; Cech, S; Kubelka, M; Lopatárová, M; Holý, L; Jindra, M

    2006-11-01

    The vitality of bovine oocytes stored in isolated follicles was examined. The aim of this work was to prolong the time of in vitro manipulation of oocytes before their maturation and develop a new alternative of oocyte "capacitation" to improve the quality of in vitro produced embryos. Follicles were dissected from the ovaries of slaughtered cows; subsequently, follicles were divided according to their diameter into three categories (2-3, 3-4 and 4-6 mm), and stored at 17-18 degrees C for 24 or 48 h in a modified tissue culture medium-199 (TCM-199) with reduced pH. After that time, the cumulus-oocyte complexes (COCs) were isolated, matured, fertilized, and embryos cultured in vitro for a total of 9 days. The percentage of total blastocysts, and hatched blastocysts developed from oocytes, initially kept ("capacitated") for 24h at 17-18 degrees C, within follicles of 3-6mm size categories, were significantly higher than that oocytes of the control [of control oocytes] (44.9 and 30.3% versus 36.2 and 20.4%, respectively). The oocytes of follicles stored for 48 h at 17-18 degrees C already had decreased developmental capacity. Interesting data were obtained when COCs of the 3-4 and 4-6 categories were additionally divided into two subgroups according to their presumed developmental history (originating from the supposed growing "fit" in contrast to the supposed regressing "unfit" follicles). The higher improvement in the rate of hatched blastocysts from 24h stored oocytes was observed only in the subgroup originated from "fit" COCs (15.3 versus 25.0%, and 20.0 versus 34.4%, in the 3-4 and 4-6mm categories, respectively). The transfer of 26 blastocysts (developed of follicles kept for 24h at 17-18 degrees C) to 26 recipient heifers resulted in 18 pregnancies. Storage of follicles at 17-18 degrees C in vitro resulted not only in recovery of higher numbers of blastocysts of better quality but also facilitated the safe transport of follicles for a long distance. The

  2. Evaluation of bovine (Bos indicus) ovarian potential for in vitro embryo production in the Adamawa plateau (Cameroon).

    Science.gov (United States)

    Kouamo, J; Dawaye, S M; Zoli, A P; Bah, G S

    2014-01-01

    An abattoir study was conducted to evaluate the ovarian potential of 201 local zebu cattle from Ngaoundere, Adamawa region (Cameroon) for in vitro embryo production (IVEP). The ovaries were excised, submerged in normal saline solution (0.9%) and transported to the laboratory for a detailed evaluation. Follicles on each ovary were counted, their diameters (Φ) measured and were grouped into 3 categories: small (Φ 8 mm). Each ovary was then sliced into a petri dish; the oocytes were recovered in Dulbecco's phosphate buffered saline, examined under a stereoscope (x10) and graded into four groups based on the morphology of cumulus oophorus cells and cytoplasmic changes of the oocytes. Grade I (GI): oocytes with more than 4 layers of bunch of compact cumulus cells mass with evenly granulated cytoplasm; grade II (GII): oocyte with at least 2-4 layers of compact cumulus cell mass with evenly granulated cytoplasm; grade III (GIII): oocyte with at least one layer of compact cumulus cell mass with evenly granulated cytoplasm; grade IV (GIV): denuded oocyte with no cumulus cells or incomplete layer of cumulus cell or expanded cells and having dark or unevenly granulated cytoplasm. The effects of both ovarian (ovarian localization, corpus luteum, size and weight of ovary) and non-ovarian factors (breed, age, body condition score (BCS) and pregnancy status of cow) on the follicular population and oocyte recovery rate were determined. There were an average of 16.75±0.83 follicles per ovary. The small, medium and large follicles were 8.39±0.60, 8.14±0.43 and 0.21±0.02 respectively. Oocyte recovery was 10.97±0.43 per ovary (65%). Oocytes graded I, II, III and IV were 3.53±0.19 (32.21%), 2.72±0.15 (24.82%), 2.24±0.15 (20.43%) and 2.47±0.20 (22.54%) respectively. The oocyte quality index was 2.26. Younger non pregnant cows having BCS of 3 and large ovaries presented higher number of follicles and oocyte quality (P quality (grade I and II) acceptable for IVEP constituted 57

  3. Evaluation of bovine (Bos indicus) ovarian potential for in vitro embryo production in the Adamawa plateau (Cameroon)

    Science.gov (United States)

    Kouamo, J.; Dawaye, S.M.; Zoli, A.P.; Bah, G.S.

    2014-01-01

    An abattoir study was conducted to evaluate the ovarian potential of 201 local zebu cattle from Ngaoundere, Adamawa region (Cameroon) for in vitro embryo production (IVEP). The ovaries were excised, submerged in normal saline solution (0.9%) and transported to the laboratory for a detailed evaluation. Follicles on each ovary were counted, their diameters (Φ) measured and were grouped into 3 categories: small (Φ 8 mm). Each ovary was then sliced into a petri dish; the oocytes were recovered in Dulbecco’s phosphate buffered saline, examined under a stereoscope (x10) and graded into four groups based on the morphology of cumulus oophorus cells and cytoplasmic changes of the oocytes. Grade I (GI): oocytes with more than 4 layers of bunch of compact cumulus cells mass with evenly granulated cytoplasm; grade II (GII): oocyte with at least 2-4 layers of compact cumulus cell mass with evenly granulated cytoplasm; grade III (GIII): oocyte with at least one layer of compact cumulus cell mass with evenly granulated cytoplasm; grade IV (GIV): denuded oocyte with no cumulus cells or incomplete layer of cumulus cell or expanded cells and having dark or unevenly granulated cytoplasm. The effects of both ovarian (ovarian localization, corpus luteum, size and weight of ovary) and non-ovarian factors (breed, age, body condition score (BCS) and pregnancy status of cow) on the follicular population and oocyte recovery rate were determined. There were an average of 16.75±0.83 follicles per ovary. The small, medium and large follicles were 8.39±0.60, 8.14±0.43 and 0.21±0.02 respectively. Oocyte recovery was 10.97±0.43 per ovary (65%). Oocytes graded I, II, III and IV were 3.53±0.19 (32.21%), 2.72±0.15 (24.82%), 2.24±0.15 (20.43%) and 2.47±0.20 (22.54%) respectively. The oocyte quality index was 2.26. Younger non pregnant cows having BCS of 3 and large ovaries presented higher number of follicles and oocyte quality (P BCS and ovarian size must be taken into account to

  4. Evaluation of bovine (Bos indicus ovarian potential for in vitro embryo production in the Adamawa plateau (Cameroon

    Directory of Open Access Journals (Sweden)

    J. Kouamo

    2014-12-01

    Full Text Available An abattoir study was conducted to evaluate the ovarian potential of 201 local zebu cattle from Ngaoundere, Adamawa region (Cameroon for in vitro embryo production (IVEP. The ovaries were excised, submerged in normal saline solution (0.9% and transported to the laboratory for a detailed evaluation. Follicles on each ovary were counted, their diameters (Φ measured and were grouped into 3 categories: small (Φ 8 mm. Each ovary was then sliced into a petri dish; the oocytes were recovered in Dulbecco’s phosphate buffered saline, examined under a stereoscope (x10 and graded into four groups based on the morphology of cumulus oophorus cells and cytoplasmic changes of the oocytes. Grade I (GI: oocytes with more than 4 layers of bunch of compact cumulus cells mass with evenly granulated cytoplasm; grade II (GII: oocyte with at least 2-4 layers of compact cumulus cell mass with evenly granulated cytoplasm; grade III (GIII: oocyte with at least one layer of compact cumulus cell mass with evenly granulated cytoplasm; grade IV (GIV: denuded oocyte with no cumulus cells or incomplete layer of cumulus cell or expanded cells and having dark or unevenly granulated cytoplasm. The effects of both ovarian (ovarian localization, corpus luteum, size and weight of ovary and non-ovarian factors (breed, age, body condition score (BCS and pregnancy status of cow on the follicular population and oocyte recovery rate were determined. There were an average of 16.75±0.83 follicles per ovary. The small, medium and large follicles were 8.39±0.60, 8.14±0.43 and 0.21±0.02 respectively. Oocyte recovery was 10.97±0.43 per ovary (65%. Oocytes graded I, II, III and IV were 3.53±0.19 (32.21%, 2.72±0.15 (24.82%, 2.24±0.15 (20.43% and 2.47±0.20 (22.54% respectively. The oocyte quality index was 2.26. Younger non pregnant cows having BCS of 3 and large ovaries presented higher number of follicles and oocyte quality (P < 0.05 compared with other animals. Oocytes with

  5. Recipient screening in IVF: First data from women undergoing anonymous oocyte donation in Dublin

    LENUS (Irish Health Repository)

    Walsh, Anthony PH

    2011-04-20

    Abstract Background Guidelines for safe gamete donation have emphasised donor screening, although none exist specifically for testing oocyte recipients. Pre-treatment assessment of anonymous donor oocyte IVF treatment in Ireland must comply with the European Union Tissues and Cells Directive (Directive 2004\\/23\\/EC). To determine the effectiveness of this Directive when applied to anonymous oocyte recipients in IVF, we reviewed data derived from selected screening tests performed in this clinical setting. Methods Data from tests conducted at baseline for all women enrolling as recipients (n = 225) in the anonymous oocyte donor IVF programme at an urban IVF referral centre during a 24-month period were analysed. Patient age at programme entry and clinical pregnancy rate were also tabulated. All recipients had at least one prior negative test for HIV, Hepatitis B\\/C, chlamydia, gonorrhoea and syphilis performed by her GP or other primary care provider before reproductive endocrinology consultation. Results Mean (±SD) age for donor egg IVF recipients was 40.7 ± 4.2 yrs. No baseline positive chlamydia, gonorrhoea or syphilis screening results were identified among recipients for anonymous oocyte donation IVF during the assessment interval. Mean pregnancy rate (per embryo transfer) in this group was 50.5%. Conclusion When tests for HIV, Hepatitis B\\/C, chlamydia, gonorrhoea and syphilis already have been confirmed to be negative before starting the anonymous donor oocyte IVF sequence, additional (repeat) testing on the recipient contributes no new clinical information that would influence treatment in this setting. Patient safety does not appear to be enhanced by application of Directive 2004\\/23\\/EC to recipients of anonymous donor oocyte IVF treatment. Given the absence of evidence to quantify risk, this practice is difficult to justify when applied to this low-risk population.

  6. Preferences of subfertile women regarding elective single embryo transfer : additional in vitro fertilization cycles are acceptable, lower pregnancy rates are not

    NARCIS (Netherlands)

    Twisk, Moniek; van der Veen, Fulco; Repping, Sjoerd; Heineman, Maas-Jan; Korevaar, Johanna C.; Bossuyt, Patrick M. M.

    2007-01-01

    With identical pregnancy rates after elective single embryo transfer (ET) and double ET strategies consisting of three cycles of IVF or intracytoplasmic sperm injection (ICSI) plus transfers of thawed/frozen embryos if available, 46% of the women undergoing IVF/ICSI favor elective single ET. If elec

  7. In-vitro maturation versus IVF with GnRH antagonist for women with polycystic ovary syndrome: treatment outcome and rates of ovarian hyperstimulation syndrome.

    Science.gov (United States)

    Das, Mausumi; Son, Weon-Young; Buckett, William; Tulandi, Togas; Holzer, Hananel

    2014-11-01

    In-vitro maturation (IVM) treatment has gained popularity for decreasing the incidence of ovarian hyperstimulation syndrome (OHSS) by eliminating or minimizing the use of gonadotrophins in women with polycystic ovary syndrome (PCOS). Studies have shown that IVF with GnRH-antagonist protocol is associated with a lower incidence of OHSS. Data comparing the relative success of these two treatments is, however, lacking. Treatment outcome and rates of OHSS were compared in patients with PCOS who underwent assisted conception with either IVM or IVF with GnRH-antagonist protocol between 2006 and 2011. The number of oocytes retrieved was higher in the IVM group, whereas the number of mature oocytes, fertilization rate and number of embryos cleaved were comparable. The implantation rate was higher in the IVF group. The clinical pregnancy rates per embryo transfer were not statistically different (IVF: 45.8% versus IVM: 32.4%). The live-birth rate was higher in the IVF group (IVF: 40.7% versus IVM: 23.5%; P = 0.04). Five women developed moderate or severe OHSS in the IVF group, whereas none did in the IVM group. Both IVM and IVF with GnRH-antagonist protocol seem to be effective treatment regimens in women with PCOS, although IVM is associated with a lower risk of OHSS.

  8. 静脉复合麻醉对IVF-ET妊娠结局的影响%Effect of intravenous mixed anesthesia on pregnancy outcome of IVF-ET

    Institute of Scientific and Technical Information of China (English)

    勾明月; 宋学茹

    2012-01-01

    Objective: To study effect of intravenous anesthesia with propofol and fentanyl on outcome of pregnancy in vitro fertilization and embryo transfer( IVF-ET). Methods: 288 patients in transvaginal oocyte retrieval of IVF/ICSI were retrospectively analyzed and the patients were divided into two groups: 216 cases of intravenous combined anesthesia group and 72 cases of no intravenous combined anesthesia group( control group). The oocyte retrieval rates, normal fertilization rates, cleavage rates,good embryo rates, embryo implantation rates, pregnancy rates, abortion rates,ive birth rates and birth defect rates were compared between the two groups in order to observe the influence of propofol and fentanyl intravenous anesthesia on outcome of pregnancy of IVF-ET. Results: There were no significant difference between the two groups in oocyte retrieval rates, normal fertilization rates,cleavage rates, good embryo rates, embryo implantation rates, pregnancy rates, abortion rates, live birth rates and birth defect rates. Conclusion: Propofol and fentanyl intravenous combined anesthesia has no effect on outcome of pregnancy of IVF-ET.%目的:探讨丙泊酚联合芬太尼静脉复合麻醉对体外受精-胚胎移植(IVF-ET)妊娠结局的影响.方法:回顾性分析静脉复合麻醉和无静脉麻醉(对照组)经阴道取卵术的体外受精/单精子卵泡浆内注射(IVF/ICSI)患者共288例,其中静脉复合麻醉组216例,对照组72例,比较两组患者获卵率、正常受精率、卵裂率、优胚率、胚胎种植率、妊娠率、流产率、活产率及出生缺陷发生率,观察丙泊酚联合芬太尼静脉复合麻醉对妊娠结局是否有影响.结果:两组患者获卵率、正常受精率、卵裂率、优胚率、胚胎种植率、妊娠率、流产率、活产率及出生缺陷发生率,差别无显著性意义.结论:丙泊酚联合芬太尼静脉复合麻醉不影响ⅣF-ET的妊娠结局.

  9. Developmental disparity between in vitro-produced and somatic cell nuclear transfer bovine days 14 and 21 embryos

    DEFF Research Database (Denmark)

    Alexopoulos, Natalie I.; Maddox-Hyttel, Poul; Tveden-Nyborg, Pernille Yde;

    2008-01-01

    , immunohistochemistry, and transmission electron microscopy to establish in vivo developmental milestones. Following morphological examination, samples were characterized for the presence of epiblast (POU5F1), mesoderm (VIM), and neuroectoderm (TUBB3). On D14, only 25, 15, and 7% of IVP, SUZI, and HMC embryos were...... of VIM and TUBB3 was less distinct in SCNT embryos when compared with IVP embryos, indicating slower or compromised development. In conclusion, SCNT and to some degree, IVP embryos displayed a high rate of embryonic mortality before D14 and surviving embryos displayed reduced quality with respect...

  10. Aberrant DNA methylation in 5'regions of DNA methyltransferase genes in aborted bovine clones

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    High rate of abortion and developmental abnormalities is thought to be closely associated with inefficient epigenetic reprogramming of the transplanted nuclei during bovine cloning.It is known that one of the important mechanisms for epigenetic reprogramming is DNA methylation.DNA methylation is established and maintained by DNA methyltransferases(DNMTs),therefore,it is postulated that the inefficient epigenetic reprogramming of transplanted nuclei may be due to abnormal expression of DNMTs.Since DNA methylation can strongly inhibit gene expression,aberrant DNA methylation of DNMT genes may disturb gene expression.But presently,it is not clear whether the methylation abnormality of DNMT genes is related to developmental failure of somatic cell nuclear transfer embryos.In our study,we analyzed methylation patterns of the 5' regions of four DNMT genes including Dnmt3a,Dnmt3b,Dnmtl and Dnmt2 in four aborted bovine clones.Using bisulfite sequencing method,we found that 3 out of 4 aborted bovine clones(AF1,AF2 and AF3)showed either hypermethylation or hypomethylation in the 5' regions of Dnmt3a and Dnmt3b.indicating that Dnmt3a and Dnmt3b genes are not properly reprogrammed.However,the individual AF4 exhibited similar methylation level and pattern to age-matched in vitro fertilized (IVF)fetuses.Besides,we found that tle 5'regions of Dnmtl and Dnmt2 were nearly completely unmethylated in all normal adults.IVF fetuses,sperm and aborted clones.Together,our results suggest that the aberrant methylation of Dnmt3a and Dnmt3b 5' regions is probably associated with the high abortion of bovine clones.

  11. The usefulness of endometrial thickness, morphology and vasculature by 2D Doppler ultrasound in prediction of pregnancy in IVF/ICSI cycles

    Directory of Open Access Journals (Sweden)

    Moustafa Abdel Kader

    2016-03-01

    Conclusion: When the endometrial thickness is <8 mm, and if there are non-triple endometrial line and non-favorable blood flow zone on day of hCG in IVF/ICSI cycles, pregnancy is unlikely and embryo transfer should be canceled with freezing of all embryos for future transfer to increase the success rate.

  12. Bivariate analysis of basal serum anti-Müllerian hormone measurements and human blastocyst development after IVF

    Directory of Open Access Journals (Sweden)

    Sills E Scott

    2011-12-01

    Full Text Available Abstract Background To report on relationships among baseline serum anti-Müllerian hormone (AMH measurements, blastocyst development and other selected embryology parameters observed in non-donor oocyte IVF cycles. Methods Pre-treatment AMH was measured in patients undergoing IVF (n = 79 and retrospectively correlated to in vitro embryo development noted during culture. Results Mean (+/- SD age for study patients in this study group was 36.3 ± 4.0 (range = 28-45 yrs, and mean (+/- SD terminal serum estradiol during IVF was 5929 +/- 4056 pmol/l. A moderate positive correlation (0.49; 95% CI 0.31 to 0.65 was noted between basal serum AMH and number of MII oocytes retrieved. Similarly, a moderate positive correlation (0.44 was observed between serum AMH and number of early cleavage-stage embryos (95% CI 0.24 to 0.61, suggesting a relationship between serum AMH and embryo development in IVF. Of note, serum AMH levels at baseline were significantly different for patients who did and did not undergo blastocyst transfer (15.6 vs. 10.9 pmol/l; p = 0.029. Conclusions While serum AMH has found increasing application as a predictor of ovarian reserve for patients prior to IVF, its roles to estimate in vitro embryo morphology and potential to advance to blastocyst stage have not been extensively investigated. These data suggest that baseline serum AMH determinations can help forecast blastocyst developmental during IVF. Serum AMH measured before treatment may assist patients, clinicians and embryologists as scheduling of embryo transfer is outlined. Additional studies are needed to confirm these correlations and to better define the role of baseline serum AMH level in the prediction of blastocyst formation.

  13. Cryo-thawed embryo transfer : natural versus artificial cycle. A non-inferiority trial. (ANTARCTICA trial)

    NARCIS (Netherlands)

    Groenewoud, Eva R.; Macklon, Nick S.; Cohlen, Ben J.

    2012-01-01

    Background: Frozen thawed embryo transfer (FET) is a cost-effective adjunct to IVF or IVF-ICSI treatment. In order to optimize treatment outcome, FET should be carried out during a period of optimal endometrial receptivity. To optimize implantation several methods for endometrium preparation have be

  14. 尿源性高纯度FSH在控制性促排卵中对IVF-ET结局的影响%Effect of domestic highly purified urinary follicle stimulating hormone on outcomes of in vitro fertilization-embryo transfer in controlled ovarian stimulation

    Institute of Scientific and Technical Information of China (English)

    叶虹; 黄国宁; 曹云霞; 钟影; 黄元华; 朱桂金; 周黎明; 陈子江; 师娟子

    2013-01-01

    Objective To investigate the effect of domestic urine-derived high-purity follicle-stimulating hormone (HP-FSH,Lishenbao) on the outcome of in vitro fertilization(IVF) embryo transfer (ET) in controlled ovarian stimulation (COS).Methods From 1 September 2010 to 31 March 2011,total of 3178 infertility patients from 14 Reproductive Center with IVF or intracytoplasmic sperm injection (ICSI) indications who accepted first IVF or ICSI cycle were studied retrospectively.Their causes of infertility include all infertility factors except ovulatory dysfunction infertility and uterine factor infertility.The only long luteal phase gonadotropin-releasing hormone agonist (GnRH-a) protocol was included.Patients were divided into 2 groups according to the type of follicle-stimulating hormone (FSH) agents used:1932 cases in HP-FSH group and 1246 cases in recombinant FSH (rFSH)group.Patients in both groups were combined with human menopausal gonadotropin (hMG) at doses of 150 U when follicle with diameter reached to 14-16 mm.When 3 dominate follicle with diameter reached 18 mm,hCG at dose of 5000 to 10 000 U orrecombinant hCG at dose of 250 μg was administered by intramuscular injection.After 34 to 36 hours,oocytes were obtained guided by ultrasound,then IVF-ET were underwent in their Reproductive Center.The primary endpoint was comparison of live birth rate between the two groups.The secondary endpoints were comparisons of clinical pregnancy rate,miscarriage rate,and implantation rate,as well as COS and IVF outcome between the two groups.Results (1) There were significantly differences in baseline characteristics of the patients between two groups.The mean age was elder(32 ±4 versus 30 ±4,P <0.01),the infertility duration was longer(5 ±4 versus 5 ± 3,P < 0.01),and antral follicle count (AFC) was less (11 ± 5 versus 13 ± 7,P < 0.01)in patients of HP-FSH group compared with those in patients of rFSH group.(2) As c ompared with rFSH,the total doses of gonadotropin needed

  15. Effect of oxygen concentration on human embryo development evaluated by time-lapse monitoring

    DEFF Research Database (Denmark)

    Ingerslev, Hans Jakob; Hindkjær, Johnny Juhl; Kirkegaard, Kirstine

    2012-01-01

    by using a statistical programme (STATA) to random choose half of the embryos from each patient, generating group 2 consisting of 98 IVF embryos from 21 patients and group 3 consisting of 109 ICSI embryos from 38 patients. Automated image recording was performed every 20 min. Key events such as time...

  16. Epidermal growth factor improves developmental competence and embryonic quality of singly cultured domestic cat embryos

    OpenAIRE

    THONGKITTIDILOK, Chommanart; THARASANIT, Theerawat; Songsasen, Nucharin; SANANMUANG, Thanida; BUARPUNG, Sirirak; TECHAKUMPHU, Mongkol

    2015-01-01

    This study examined the influence of EGF on the expression of EGF receptors (EGFR) and developmental competence of embryos cultured individually versus those cultured in groups. Cat oocytes were in vitro matured and fertilized (IVM/IVF), and cleaved embryos were randomly assigned to one of seven culture conditions: one group each in which embryos were subjected to group culture supplemented with or without 5 ng/ml EGF and five groups in which embryos were subjected to single-embryo culture su...

  17. Factors influencing success rate of IVF/ET:analysis of 770 treatment cycles

    Institute of Scientific and Technical Information of China (English)

    叶碧绿; 林金菊; 周颖; 黄学峰; 赵军招; 郑菊芬; 林文琴

    2002-01-01

    Objective: To evaluate the factors influencing success ra te of in vitro fertilization /embryo transfer(IVF/ET).Methods: A retrospective study of 770 consecutive cycles under gone IVF/ET or intracytoplasmic sperm injection (ICSI) from March 1999 to June 2 001.Estradiol and progesterone concentrations on the day of hCG administration w ere measured. Factors effecting embryo transfer on the effects of clinical pregn ancy rate were evaluated.Results: The overall clinical pregnancy rate in 770 cycles was 40.8%. Take home baby rate was 31%. No significantly differences rate were obse rved between groups with different estradiol levels.If the progesterone concentr ation on the day of hCG administration was >6.36 nmol/L, the implantation and cl inical pregnancy rate was lower(P<0.003). The lowest clinical pregnancy rat e was observed when the duration of infertility was ≥10 years. The average numb er of embryos transferred was 2.23±0.83.Conclusion: This study demonstrated that the most important fac tor on clinical pregnancy rate was low progesterone concentration on the day of hCG administration. The other two important variables influencing success rate o f IVF/ET were the duration of infertility, the number of high-grade embryos tra nsferred.

  18. Gonadotropin stimulation regimens for follicular aspiration and in vitro embryo production from calf oocytes.

    Science.gov (United States)

    Armstrong, D T; Irvine, B J; Earl, C R; McLean, D; Seamark, R F

    1994-01-01

    Crossbred beef x dairy calves were randomly allocated at 3 wk of age to different gonadotropin treatment regimens for stimulation of follicle development and induction of oocyte maturation in vivo. Follicular responses were assessed laparoscopically, and oocytes were aspirated for assessment of maturational state or for in vitro fertilization (IVF) and culture to determine developmental capacity. Follicle-stimulating Hormone (FSH), administered in a single subcutaneous injection together with a low dosage of PMSG, was as effective as the same total dosage of FSH administered in 6 injections over a 3-d period. Without accompanying PMSG, this dose of FSH was ineffective in stimulating follicle development. The mean number of preovulatory follicles (> 5mm, with hyperemic appearance) doubled with each successive stimulation at 3-wk intervals, reaching 35 follicles per calf at 9 wk of age. Oocyte yields ranged from 55 to 81% of follicles aspirated, and did not differ significantly among age, FSH regimen and oocyte maturation stimulus. A combination of LH + FSH was more effective in stimulating cumulus cell expansion than LH by itself (73 vs 22% of recovered oocyte-cumulus cell complex (OCC) respectively; Poocytes (cumulus unexpanded) subjected to in vitro maturation (IVM) and IVF, 30% developed to blastocysts during co-culture with bovine oviduct epithelial cells, which was not significantly different from 25% of 36 oocytes from adult ovaries which reached the blastocyst stage under similar conditions. The results indicate that follicle responses of calf ovaries to FSH stimulation increase progressively from 3 to 9 wk of age, and that oocytes recovered laparoscopically from these follicles produce blastocysts in culture at rates similar to oocytes from adult cattle ovaries collected at slaughter. The approach offers promise for embryo production from donor calves of superior genetic merit for embryo transfer, thereby enhancing the rate of genetic gain above that

  19. Granulocyte colony-stimulating factor in repeated IVF failure, a randomized trial.

    Science.gov (United States)

    Aleyasin, Ashraf; Abediasl, Zhila; Nazari, Atefeh; Sheikh, Mahdi

    2016-06-01

    Recent studies have revealed key roles for granulocyte colony-stimulating factor (GCSF) in embryo implantation process and maintenance of pregnancy, and some studies showed promising results by using local intrauterine infusion of GCSF in patients undergoing in vitro fertilization (IVF). This multicenter, randomized, controlled trial included 112 infertile women with repeated IVF failure to evaluate the efficacy of systemic single-dose subcutaneous GCSF administration on IVF success in these women. In this study, the Long Protocol of ovarian stimulation was used for all participants. Sealed, numbered envelopes assigned 56 patients to receive subcutaneous 300 µg GCSF before implantation and 56 in the control group. The implantation (number of gestational sacs on the total number of transferred embryos), chemical pregnancy (positive serum β-HCG), and clinical pregnancy (gestational sac and fetal heart) rates were compared between the two groups. This trial is registered at www.irct.ir (IRCT201503119568N11). The successful implantation (18% vs 7.2%, P=0.007), chemical pregnancy (44.6% vs 19.6%, P=0.005), and clinical pregnancy (37.5% vs 14.3%, P=0.005) rates were significantly higher in the intervention group than in the control group. After adjustment for participants' age, endometrial thickness, good-quality oocyte counts, number of transferred embryos, and anti-Mullerian hormone levels, GCSF treatment remained significantly associated with successful implantation (OR=2.63, 95% CI=1.09-6.96), having chemical pregnancy (OR= 2.74, 95% CI=1.11-7.38) and clinical pregnancy (OR=2.94, 95% CI=1.23-8.33). In conclusion, administration of single-dose systemic subcutaneous GCSF before implantation significantly increases the IVF success, implantation, and pregnancy rates in infertile women with repeated IVF failure. PMID:26980809

  20. Making Muslim babies: IVF and gamete donation in Sunni versus Shi'a Islam.

    Science.gov (United States)

    Inhorn, Marcia C

    2006-12-01

    Medical anthropological research on science, biotechnology, and religion has focused on the "local moral worlds" of men and women as they make difficult decisions regarding their health and the beginnings and endings of human life. This paper focuses on the local moral worlds of infertile Muslims as they attempt to make, in the religiously correct fashion, Muslim babies at in vitro fertilization (IVF) clinics in Egypt and Lebanon. As early as 1980, authoritative fatwas issued from Egypt's famed Al-Azhar University suggested that IVF and similar technologies are permissible as long as they do not involve any form of third-party donation (of sperm, eggs, embryos, or uteruses). Since the late 1990s, however, divergences in opinion over third-party gamete donation have occurred between Sunni and Shi'ite Muslims, with Iran's leading ayatollah permitting gamete donation under certain conditions. This Iranian fatwa has had profound implications for the country of Lebanon, where a Shi'ite majority also seeks IVF services. Based on three periods of ethnographic research in Egyptian and Lebanese IVF clinics, this paper explores official and unofficial religious discourses surrounding the practice of IVF and third-party donation in the Muslim world, as well as the gender implications of gamete donation for Muslim marriages. PMID:17051430

  1. Effect of antral follicle count (AFC) in beef heifers on in vitro fertilization/production (IVF/IVP)

    Science.gov (United States)

    Our objective has been to compare the in vitro fertilization (IVF) and production (IVP) of embryos from low and high antral follicle count (AFC) heifers. This is the 4th year of the study with years 1 to 3 reported individually. For this report, we add data for the 4th year and present a combined an...

  2. In vitro fertilization (IVF) using semi-defined culture conditions from low or high antral follicle count pubertal beef heifers

    Science.gov (United States)

    To compare the in vitro fertilization (IVF) and production (IVP) of embryos from low and high antral follicle count (AFC) heifers, AFC were determined on 106 heifers using transrectal ultrasonography. Ten heifers with the lowest AFC (avg. 13.2) and 10 heifers with the highest AFC (avg. 27.4) with ev...

  3. 牛与小鼠胚胎在分化抑制培养系统中的行为比较%Comparison of the growth behavior between bovine and murine embryo in a differentiation inhibitory system

    Institute of Scientific and Technical Information of China (English)

    杨奇; 安立龙; 窦忠英; 高志敏; 雷安民; 杨春荣

    2001-01-01

    在自制培养基中,以原代小鼠胎儿成纤维细胞为饲养层,观察比较了牛与小鼠胚胎在体外分化抑制培养体系中的生长行为。结果表明,牛囊胚一般培养36~60 h后孵化脱带,96~120 h后贴壁,120~144 h为ICM传代的最佳时刻。小鼠囊胚一般培养12~24 h孵化脱带,24~48 h贴壁,72~96 h为传代的最佳时刻。牛胚胎贴附于饲养层上生长,极易从饲养层上剥离;小鼠胚胎镶嵌在饲养层中,滋养层细胞与饲养层细胞间连接紧密。牛ICM色深发黑,集落隆起程度较低;小鼠ICM呈暗黄色,有呈柱状增殖的趋势。牛滋养层呈网状,小鼠滋养层为较致密的单层薄膜。%The growth behavior differences between bovine embryos and murine embryos were studied on PMEF feeder layer using DMEM+15% NBS+0.1 mmol/L β-mercaptoetheanol+0.1 μmol/L Na2 SeO3+10 μg/mL LIF+10 μg/mL IGF-1 as the media.The results showed as follows:In the culture system,bovine blastocyst hatched affer being cultured for 36-60 h,and attached on the feeder layer after being cultured for96-120 h.After cultured for 120-144 h,the growing ICM were passaged at first.Morulae attached on PMEF feeder layer 12-24 h later than blastocyst for passing on blastula stage and hatching period.Murine blastocyst hatched after being cultured for 12-24 h,and attached on the feeder layer after cultured for 24-48 h.After cultured for 72-96 h,the growing ICM were passaged.Bovine embryo lived on the feeder layer and easy to be stripped from the feeder layer,while murine embryo implanted in the feeder layer,and murine trophoblastic cells were closely connected with feeder layer cells arrounding them.The color of bovine ICM was more black than murine ICM.However,the colonies of bovine ICM were lower compared with murine ICM colonies,and murine ICM tended to be column-like.Bovine trophoblast was net-like,and murine trophoblast formed a membranous structure.

  4. An endometrial gene expression signature accurately predicts recurrent implantation failure after IVF

    Science.gov (United States)

    Koot, Yvonne E. M.; van Hooff, Sander R.; Boomsma, Carolien M.; van Leenen, Dik; Groot Koerkamp, Marian J. A.; Goddijn, Mariëtte; Eijkemans, Marinus J. C.; Fauser, Bart C. J. M.; Holstege, Frank C. P.; Macklon, Nick S.

    2016-01-01

    The primary limiting factor for effective IVF treatment is successful embryo implantation. Recurrent implantation failure (RIF) is a condition whereby couples fail to achieve pregnancy despite consecutive embryo transfers. Here we describe the collection of gene expression profiles from mid-luteal phase endometrial biopsies (n = 115) from women experiencing RIF and healthy controls. Using a signature discovery set (n = 81) we identify a signature containing 303 genes predictive of RIF. Independent validation in 34 samples shows that the gene signature predicts RIF with 100% positive predictive value (PPV). The strength of the RIF associated expression signature also stratifies RIF patients into distinct groups with different subsequent implantation success rates. Exploration of the expression changes suggests that RIF is primarily associated with reduced cellular proliferation. The gene signature will be of value in counselling and guiding further treatment of women who fail to conceive upon IVF and suggests new avenues for developing intervention. PMID:26797113

  5. The effect of intrauterine HCG injection on IVF outcome: a systematic review and meta-analysis.

    Science.gov (United States)

    Osman, A; Pundir, J; Elsherbini, M; Dave, S; El-Toukhy, T; Khalaf, Y

    2016-09-01

    In this systematic review and meta-analysis, the effect of intrauterine HCG infusion before embryo transfer on IVF outcomes (live birth rate, clinical pregnancy rate and spontaneous aboretion rate) was investigated. Searches were conducted on MEDLINE, EMBASE and The Cochrane Library. Randomized studies in women undergoing IVF and intracytoplasmic sperm injection comparing intrauterine HCG administration at embryo transfer compared with no intrauterine HCG were eligible for inclusion. Eight randomized controlled trials were eligible for inclusion in the meta-analysis. A total of 3087 women undergoing IVF and intracytoplasmic sperm injection cycles were enrolled (intrauterine HCG group: n = 1614; control group: n = 1473). No significant difference was found in the live birth rate (RR 1.13; 95% CI 0.84 to 1.53) and spontaneous abortion rate (RR 1.00, 95% CI 0.74 to 1.34) between women who received intrauterine HCG and those who did not receive HCG. Although this review was extensive and included randomized controlled trials, no significant heterogeneity was found, and the overall included numbers are relatively small. In conclusion the current evidence does not support the use of intrauterine HCG administration before embryo transfer. Well-designed multicentre trials are needed to provide robust evidence.

  6. Cryopreservation of manipulated embryos: tackling the double jeopardy.

    Science.gov (United States)

    Dinnyes, A; Nedambale, T L

    2009-01-01

    The aim of the present review is to provide information to researchers and practitioners concerning the reasons for the altered viability and the medium- and long-term consequences of cryopreservation of manipulated mammalian embryos. Embryo manipulation is defined herein as the act or process of manipulating mammalian embryos, including superovulation, AI, IVM, IVF, in vitro culture, intracytoplasmic sperm injection, embryo biopsy or splitting, somatic cell nuclear transfer cloning, the production of sexed embryos (by sperm sexing), embryo cryopreservation, embryo transfer or the creation of genetically modified (transgenic) embryos. With advances in manipulation technologies, the application of embryo manipulation will become more frequent; the proper prevention and management of the resulting alterations will be crucial in establishing an economically viable animal breeding technology.

  7. Does local endometrial injury in the nontransfer cycle improve the IVF-ET outcome in the subsequent cycle in patients with previous unsuccessful IVF? A randomized controlled pilot study

    Directory of Open Access Journals (Sweden)

    Sachin A Narvekar

    2010-01-01

    Full Text Available Background: Management of repeated implantation failure despite transfer of good-quality embryos still remains a dilemma for ART specialists. Scrapping of endometrium in the nontransfer cycle has been shown to improve the pregnancy rate in the subsequent IVF/ET cycle in recent studies. Aim: The objective of this randomized controlled trial (RCT was to determine whether endometrial injury caused by Pipelle sampling in the nontransfer cycle could improve the probability of pregnancy in the subsequent IVF cycle in patients who had previous failed IVF outcome. Setting: Tertiary assisted conception center. Design: Randomized controlled study. Materials and Methods: 100 eligible patients with previous failed IVF despite transfer of good-quality embryos were randomly allocated to the intervention group and control groups. In the intervention group, Pipelle endometrial sampling was done twice: One in the follicular phase and again in the luteal phase in the cycle preceding the embryo transfer cycle. Outcome Measure: The primary outcome measure was live birth rate. The secondary outcome measures were implantation and clinical pregnancy rates. Results: The live birth rate was significantly higher in the intervention group compared to control group (22.4% and 9.8% P = 0.04. The clinical pregnancy rate in the intervention group was 32.7%, while that in the control group was 13.7%, which was also statistically significant ( P = 0.01. The implantation rate was significantly higher in the intervention group as compared to controls (13.07% vs 7.1% P = 0.04. Conclusions: Endometrial injury in nontransfer cycle improves the live birth rate,clinical pregnancy and implantation rates in the subsequent IVF-ET cycle in patients with previous unsuccessful IVF cycles.

  8. Efficacy of natural cycle IVF : a review of the literature

    NARCIS (Netherlands)

    Pelinck, MJ; Hoek, A; Simons, AHM; Heineman, MJ

    2002-01-01

    Since the introduction of IVF treatments, natural cycle IVF has been largely replaced by IVF with ovarian stimulation. However, natural cycle IVF has several advantages. It is associated with a close to zero multiple pregnancy rate, and a zero risk of ovarian hyperstimulation syndrome. Per cycle, na

  9. Children conceived by in vitro fertilisation after fresh embryo transfer

    OpenAIRE

    D'Souza, S; Rivlin, E; Cadman, J; Richards, B; Buck, P; Lieberman, B.

    1997-01-01

    AIMS—To compare the outcome in in vitro fertilisation (IVF) children (after fresh embryo transfer) from multiple and singleton births with one another, and with normally conceived control children.
METHODS—A cohort of 278 children (150 singletons, 100 twins, 24 triplets and four quadruplets), conceived by IVF after three fresh embryos had been transferred, born between October 1984 and December 1991, and 278 normally conceived control children (all singletons), were followed up for four years...

  10. Embryo disposition and the new death scene

    Directory of Open Access Journals (Sweden)

    Ellison, David

    2011-01-01

    Full Text Available In the IVF clinic - a place designed principally for the production and implantation of embryos - scientists and IVF recipients are faced with decisions regarding the disposition of frozen embryos. At this time there are hundred of thousands of cryopreserved embryos awaiting such determinations. They may be thawed for transfer to the woman herself, they may be donated for research or for use by other infertile couples, they may remain in frozen storage, or they may variously be discarded by being allowed to 'succumb', or 'perish'. Where the choice is discard, some IVF clients have chosen to formalise the process through ceremony. A new language is emerging in response to the desires of the would-be-parents who might wish to characterise the discard experience as a ‘good death’. This article examines the procedure known as ‘compassionate transfer’ where the embryo to be discarded is placed in the woman’s vagina where it is clear that it will not develop further. An alternate method has the embryo transferred in the usual manner but without the benefit of fertility-enhancing hormones at a point in the cycle unreceptive to implantation. The embryo destined for disposal is thus removed from the realm of technological possibility and ‘returned’ to the female body for a homely death. While debates continue about whether or not embryos constitute life, new practices are developing in response to the emotional experience of embryo discard. We argue that compassionate transfer is a death scene taking shape. In this article, we take the measure of this new death scene’s fabrication, and consider the form, significance, and legal complexity of its ceremonies.

  11. The German IVF-Registry – D·I·R

    Directory of Open Access Journals (Sweden)

    Bühler K

    2013-01-01

    Full Text Available Systematic data collection in the field of assisted reproductive techniques is performed in Germany since 30 years. Since 1997, data collection is performed electronically and in a prospective way. In 1998, by a code of the German Medical Association participation at the registry has become mandatory for all IVF centres in Germany. Different software solutions can be used in the centres for data collection. All of them have to cooperate with a special “DIRmod-DLL”, controlling online plausibility and prospectivity of the collected data. More than one million treatment cycles have been reported in the annuals published since 1996 up to 2010. All in all 1.2 millions cycles are included in the registry. Clinical pregnancy rate increased continuously and miscarriage rate decreased. By reduction of the mean number of transferred embryos of about 20% the percentage of born triplets in relationship to all children born after ART decreased of about 80%. It can be shown with the huge number of documented cycles that political decisions exert a dominating influence on treatment and drug choice. The German IVF registry has been established over the last 30 years as an instrument of quality assurance and has experienced highest regard by colleagues and by the international scientific world.

  12. Follicular Proinflammatory Cytokines and Chemokines as Markers of IVF Success

    Directory of Open Access Journals (Sweden)

    Aili Sarapik

    2012-01-01

    Full Text Available Cytokines are key modulators of the immune system and also contribute to regulation of the ovarian cycle. In this study, Bender MedSystems FlowCytomix technology was used to analyze follicular cytokines (proinflammatory: IL-1β, IL-6, IL-18, IFN-γ, IFN-α, TNF-α, IL-12, and IL-23;, and anti-inflammatory: G-CSF, chemokines (MIP-1α, MIP-1β, MCP-1, RANTES, and IL-8, and other biomarkers (sAPO-1/Fas, CD44(v6 in 153 women undergoing in vitro fertilization (IVF. Cytokine origin was studied by mRNA analysis of granulosa cells. Higher follicular MIP-1α and CD44(v6 were found to correlate with polycystic ovary syndrome, IL-23, INF-γ, and TNF-α with endometriosis, higher CD44(v6 but lower IL-β and INF-α correlated with tubal factor infertility, and lower levels of IL-18 and CD44(v6 characterized unexplained infertility. IL-12 positively correlated with oocyte fertilization and embryo development, while increased IL-18, IL-8, and MIP-1β were associated with successful IVF-induced pregnancy.

  13. Obstetric complications in women with IVF conceived pregnancies and polycystic ovarian syndrome

    Directory of Open Access Journals (Sweden)

    Sunita R Tandulwadkar

    2014-01-01

    Full Text Available Polycystic ovarian syndrome (PCOS is often accompanied by infertility that necessitates ovulation induction using clomiphene citrate, gonadotropins or even in vitro fertilization (IVF. These treatment methods are known to increase the incidence of multiple pregnancies as well as some negative consequences, including a rise in the risk for gestational diabetes mellitus, pre-eclampsia, etc., Furthermore, pregnancies established after IVF carry an increased risk for maternal complications. However, the increased risk of developing adverse obstetric complications has been suggested to occur independently of obesity as well as in populations without assisted reproductive techniques. Many studies have been performed to study the effect of PCOS on pregnancy and the effect of pregnancy on PCOS. The hormonal milieu that is exaggerated in PCOS women is quite well understood at the biochemical and genetic levels. The maternal and neonatal outcomes of PCOS women who have undergone in vitro fertilization-embryo transfer (IVF-ET have not been widely studied till date. This review aims to evaluate the current evidence regarding adverse obstetric outcomes of PCOS women undergoing IVF-ET. The rationale of this review is to study whether the adverse obstetric outcomes are increased in PCOS women in general, or particularly in those PCOS women who are undergoing IVF-ET. It is also important to analyze via a literature review whether the increased adverse outcomes are due to infertility in general or PCOS per se. An attempt has been made to give evidence regarding preventive strategies for obstetric complications in PCOS women who have undergone IVF-ET.

  14. Aromatase inhibitors in stimulated IVF cycles

    DEFF Research Database (Denmark)

    Papanikolaou, Evangelos G; Polyzos, Nikolaos P; Al Humaidan, Peter Samir Heskjær;

    2011-01-01

    are available regarding their efficacy in IVF stimulated cycles. Current available evidence support that letrozole may have a promising role in stimulated IVF cycles, either when administered during the follicular phase for ovarian stimulation. Especially for women with poor ovarian response, letrozole appears...... to test in the future how it may perform when used in GnRH agonist cycles. Finally administration of letrozole during luteal phase in IVF cycles offers another treatment modality for patients at high risk for OHSS taking into account that it drastically reduces estradiol levels....

  15. GnRH拮抗剂对体外受精-胚胎移植中多囊卵巢综合征患者有效性的系统评价%Effectiveness of GnRH Antagonist In Vitro Fertilization-Embryo Transfer (IVF-ET) in PCOS Patients: A Systematic Review

    Institute of Scientific and Technical Information of China (English)

    肖劲松; 陈双陨; 张春莲; 常双

    2011-01-01

    目的 系统评价GnRH拮抗剂用于多囊卵巢综合征(PCOS)患者体外受精-胚胎移植的有效性.方法 计算机检索PubMed(1997~2010)、ProQust Medical Library(1997~2010)、外文生物医学期刊文献数据库(2000~2010)、CBM(1979~2010)、CNKI(1994~2010)、VIP(1989~2010)、万方数据库(1994~2010)和读秀学术搜索引擎,并手工检索相关中文杂志9种,查找比较GnRH拮抗剂与GnRH激动剂在体外受精-胚胎移植PCOS患者治疗中有效性的随机对照试验(RCT).由两位研究者按纳入排除标准筛选文献、提取资料和评价质量后,采用RevmMan4.2.7软件进行Meta分析.结果 共纳人6篇RCT,合计699例患者.其纳人研究质量A级3篇,B级3篇.Meta分析结果显示:与GnRH激动剂组比较,GnRH拮抗剂组在刺激天数[WMD=-1.23(-2.76,-0.31)]、Gn量[WMD=-4.87(-14.20,4.46)]、HCG日E2值[WMD=31.37(-263.40,-326)]、获卵数[WMD=1.34(-1.02,4.70)]、临床妊娠率[OR=1.27(0.77,2.10)]和流产率[PetoOR=0.67(0.38,1.18)]方面,两组差异均无统计学意义;仅GnRH拮抗剂组的OHSS率较低,两组差异有统计学意义[PetoOR=0.35(0.24,0.50)].结论 GnRH拮抗剂方案较之GnRH激动剂方案能减低OHSS发生,而在Gn量、获卵数、临床妊娠率方面,两种方案疗效相当.由于GnRH拮抗剂方案可缩短治疗时间,减少治疗费用,具有较好的安全性,对PCOS患者行体外受精-胚胎移植可能是较理想的选择.鉴于纳入研究存在质量和数量的不足以及方法学的差异,上述结论仅供临床参考,需要开展更多后效评价和不断更新.%Objective To evaluate the effectiveness of GnRH antagonist in vitro fertilization-embryo transfer (IVFET) in polycystic ovary syndrome (PCOS) patients. Methods Such databases as PubMed (1997 to 2010), PML (1997 to 2010), FMJS (2000 to 2010), CBMdisc (1979 to 2010), CNKI (1994 to 2010), VIP (1989 to 2010), WanFang (1994 to 2010), and duxiu scholar searcher (www.duxiu.com), and nine relevant

  16. [Use of an in vitro model in bovine to evidence a functional and molecular dialogue between preimplantation embryo and oviduct epithelial cells].

    Science.gov (United States)

    Cordova, A; Perreau, C; Schmaltz-Panneau, B; Locatelli, Y; Ponsart, C; Mermillod, P

    2013-09-01

    Beyond being a pipe between ovary and uterus, the oviduct is an active player in different aspects of early reproductive processes, in particular in the transport of embryos to the site of implantation and the regulation of its early development. Different studies evidenced a communication between oviduct and early embryo at the molecular and functional levels. Since the study of these interactions is difficult in vivo, different in vitro systems have been developed to mimic the maternal milieu during early development. These systems allowed to confirm the action of the cells on the quality of early development (blastocyst rate and viability). In turn, the embryos are producing signals that are able to modify and adapt the activity of maternal cells. PMID:23958329

  17. Cryopreservation of Embryos and Oocytes in Human Assisted Reproduction

    Directory of Open Access Journals (Sweden)

    János Konc

    2014-01-01

    Full Text Available Both sperm and embryo cryopreservation have become routine procedures in human assisted reproduction and oocyte cryopreservation is being introduced into clinical practice and is getting more and more widely used. Embryo cryopreservation has decreased the number of fresh embryo transfers and maximized the effectiveness of the IVF cycle. The data shows that women who had transfers of fresh and frozen embryos obtained 8% additional births by using their cryopreserved embryos. Oocyte cryopreservation offers more advantages compared to embryo freezing, such as fertility preservation in women at risk of losing fertility due to oncological treatment or chronic disease, egg donation, and postponing childbirth, and eliminates religious and/or other ethical, legal, and moral concerns of embryo freezing. In this review, the basic principles, methodology, and practical experiences as well as safety and other aspects concerning slow cooling and ultrarapid cooling (vitrification of human embryos and oocytes are summarized.

  18. Efeito do ibuprofeno administrado uma hora antes da inovulação de embriões bovinos Effect of ibuprofen administered one hour before the bovine embryo transfer

    Directory of Open Access Journals (Sweden)

    H.J. Narváez

    2010-06-01

    Full Text Available Avaliou-se o efeito do ibuprofeno administrado uma hora antes da inovulação de embriões bovinos, com o objetivo de melhorar a taxa de prenhez. Após a avaliação da resposta ao protocolo de sincronização do estro, 76 fêmeas selecionadas como receptoras de embriões foram distribuídas em três grupos (G experimentais: G1 (n=25 receptoras usadas como controle, G2 (n=30 receptoras que receberam ibuprofeno 5mg/kg, I.M, uma hora antes da inovulação dos embriões, e G3 (n=21 receptoras que receberam uma matriz polimérica de liberação controlada de ibuprofeno administrado por via subcutânea. As taxas de prenhez foram de 16% (4/25, 43,3% (13/30 e 14,2% (3/21, para G1, G2 e G3, respectivamente. Observou-se diferença (PThe effect of the administered ibuprofen was evaluated one hour before the embryo transfer of bovine embryos in order to improve pregnancy rates. After evaluating the response to protocol synchronization of estrus, 76 Females selected as the recipients of embryos were distributed into three experimental groups: G1 (n = 25 surrogate cows used as control, G2 (n = 30 surrogate cows that received 5mg/kg ibuprofen, IM, one hour before the embryo transfer, and G3 (n = 20 surrogate cows that received an array polymeric release of controlled ibuprofen subcutaneously administered. The pregnancy rates were 16% (4/25, 43.3% (13/30, and 14.2% (3/21 for G1, G2, and G3, respectively. There was statistical difference (P<0.024 on pregnancy rate of G2, in comparison with those of G1 and G3. The administration of ibuprofen intramuscularly one hour before the embryo transfer resulted in better pregnancy rate in Nellore surrogate cows.

  19. Fertilization of IVF/ICSI Using Sibling Oocytes from Couples with Subfertile Male or Unexplained Infertility

    Institute of Scientific and Technical Information of China (English)

    李志凌; 林虹; 肖婉芬; 王玉莲

    2004-01-01

    The significance of the performance of conventional in vitro fertilization and intracytoplasmic sperm injection (IVF/ICSI) using sibling oocytes from couples with subfertile male or unexplained infertility was evaluated. A total of 410 sibling oocyte cumulus-corona complexes (OCCC)from 21 couples with subfertile male (group A) and 11 unexplained infertile couples (group B) were randomly divided, in order of retrieval, into two groups inseminated either by conventional IVF or by ICSI. The treatment outcomes and the influence of infertility factors on fertilization in each group were compared. The results showed that although the two pronuclear (2PN) fertilization rate per injected sibling oocytes was significantly higher after ICSI (group A: 68.2 % ±28.8 %; group B: 66.2 %±24.9 %) than after conventional IVF (group A: 41.8 %±32.7 %; group B: 40. 1 %±22.1 %), the other variables studied included: the fertilization rates of per allocated sibling oocytes IVF/ICSI, the fertilization rates of sibling oocytes IVF/ICSI after excluding failed IVF fertilization cycles, as well as the cleavage rates of normal fertilization were not statistically significant (P>0.05). Similarly, though the total fertilization failure rate in the IVF group (group A: 42.9 %;group B: 36.4 %) was significantly higher than in the ICSI group (group A: 4.8 %; group B: 0),we did not cancel cycles due to the normal fertilization of sibling oocytes. Embryo transfer was possible in all 32 couples. There were 10 clinical pregnancies in the two groups. We also discovered a possible association between some semen parameters and sperm functions of group A, and women age and duration of infertility of group B and fertilization. It is suggested that adoption of the split IVF/ICSI technology in the above cases may help eliminate fertilization failures. This is also a useful method to investigate the effect of single factor on the employment of assisted reproductive tech nology.

  20. Sourcing human embryos for embryonic stem cell lines: Problems & perspectives

    Directory of Open Access Journals (Sweden)

    Rajvi H Mehta

    2014-01-01

    Full Text Available The ability to successfully derive human embryonic stem cells (hESC lines from human embryos following in vitro fertilization (IVF opened up a plethora of potential applications of this technique. These cell lines could have been successfully used to increase our understanding of human developmental biology, transplantation medicine and the emerging science of regenerative medicine. The main source for human embryos has been ′discarded′ or ′spare′ fresh or frozen human embryos following IVF. It is a common practice to stimulate the ovaries of women undergoing any of the assisted reproductive technologies (ART and retrieve multiple oocytes which subsequently lead to multiple embryos. Of these, only two or maximum of three embryos are transferred while the rest are cryopreserved as per the decision of the couple. In case a couple does not desire to ′cryopreserve′ their embryos then all the embryos remaining following embryo transfer can be considered ′spare′ or if a couple is no longer in need of the ′cryopreserved′ embryos then these also can be considered as ′spare′. But, the question raised by the ethicists is, "what about ′slightly′ over-stimulating a woman to get a few extra eggs and embryos? The decision becomes more difficult when it comes to ′discarded′ embryos. As of today, the quality of the embryos is primarily assessed based on morphology and the rate of development mainly judged by single point assessment. Despite many criteria described in the literature, the quality assessment is purely subjective. The question that arises is on the decision of ′discarding′ embryos. What would be the criteria for discarding embryos and the potential ′use′ of ESC derived from the ′abnormal appearing′ embryos? This paper discusses some of the newer methods to procure embryos for the derivation of embryonic stem cell lines which will respect the ethical concerns but still provide the source material.

  1. Cryopreservation of human embryos and its contribution to in vitro fertilization success rates.

    Science.gov (United States)

    Wong, Kai Mee; Mastenbroek, Sebastiaan; Repping, Sjoerd

    2014-07-01

    Cryopreservation of human embryos is now a routine procedure in assisted reproductive technologies laboratories. There is no consensus on the superiority of any protocol, and substantial differences exist among centers in day of embryo cryopreservation, freezing method, selection criteria for which embryos to freeze, method of embryo thawing, and endometrial preparation for transfer of frozen-thawed embryos. In the past decade, the number of frozen-thawed embryo transfer cycles per started in vitro fertilization (IVF) cycle increased steadily, and at the same time the percentage of frozen-thawed embryo transfers that resulted in live births increased. Currently, cryopreservation of human embryos is more important than ever for the cumulative pregnancy rate after IVF. Interestingly, success rates after frozen-thawed embryo transfer are now nearing the success rates of fresh embryo transfer. This supports the hypothesis of so called freeze-all strategies in IVF, in which all embryos are frozen and no fresh transfer is conducted, to optimize success rates. High-quality randomized controlled trials should be pursued to find out which cryopreservation protocol is best and whether the time has come to completely abandon fresh transfers.

  2. Reduced developmental competence of immature, in-vitro matured and postovulatory aged mouse oocytes following IVF and ICSI

    Directory of Open Access Journals (Sweden)

    Trounson Alan

    2008-12-01

    Full Text Available Abstract Background The present study highlights basic physiological differences associated with oocyte maturation and ageing. The study explores the fertilizing capacity and resistance to injury of mouse oocytes at different stages of maturation and ageing following IVF and ICSI. Also, the study examines the developmental competence of embryos obtained from these oocytes. The outcome of the study supports views that the mouse can be a model for human IVF suggesting that utilizing in-vitro matured and failed fertilized oocytes to produce embryos mainly when limited number of oocytes is retrieved in a specific cycle, should be carefully considered. Methods Hybrid strain mouse oocytes were inseminated by in-vitro fertilization (IVF or intracytoplasmic sperm injection (ICSI. Oocytes groups that were used were germinal vesicle (GV in-vitro matured metaphase II (IVM-MII, freshly ovulated MII (OV-MII, 13 hrs in-vitro aged MII (13 hrs-MII and 24 hrs in-vitro aged MII (24 hrs-MII. Fertilization and embryo development to the blastocyst stage were monitored up to 5 days in culture for IVF and ICSI zygotes. Sperm head decondensation and pronuclear formation were examined up to 9 hrs in oocytes following ICSI. Apoptotic events in blocked embryos were examined using the TUNNEL assay. Differences between females for the number and quality of GV and OV-MII oocytes were examined by ANOVA analyses. Differences in survival after ICSI, fertilization by IVF and ICSI and embryo development were analysed by Chi-square test with Yates correction. Results No differences in number and quality of oocytes were identified between females. The findings suggest that inability of GV oocytes to participate in fertilization and embryo development initiates primarily from their inability to support initial post fertilization events such as sperm decondensation and pronuclei formation. These events occur in all MII oocytes in similar rates (87–98% for IVF and ICSI. Following

  3. A role for Aurora C in the chromosomal passenger complex during human preimplantation embryo development

    NARCIS (Netherlands)

    Santos, Margarida Avo; van de Werken, Christine; de Vries, Marieke; Jahr, Holger; Vromans, Martijn J. M.; Laven, Joop S. E.; Fauser, Bart C.; Kops, Geert J.; Lens, Susanne M.; Baart, Esther B.

    2011-01-01

    BACKGROUND: Human embryos generated by IVF demonstrate a high incidence of chromosomal segregation errors during the cleavage divisions. To analyse underlying molecular mechanisms, we investigated the behaviour of the chromosomal passenger complex (CPC) in human oocytes and embryos. This important m

  4. Nucleolar re-activation is delayed in mouse embryos cloned from two different cell lines

    DEFF Research Database (Denmark)

    Svarcova, Olga; Dinnyes, A.; Polgar, Z.;

    2009-01-01

    Aim of this study was to evaluate and compare embryonic genome activation (EGA) in mouse embryos of different origin using nucleolus as a marker. Early and late 2-cell and late 4-cell stage embryos, prepared by in vitro fertilization (IVF), parthenogenetic activation (PG), and nuclear transfer...

  5. Early aberrations in chromatin dynamics in embryos produced under In vitro conditions

    DEFF Research Database (Denmark)

    Deshmukh, Rahul Shahaji; Østrup, Olga; Strejcek, Frantisek;

    2012-01-01

    In vitro production of porcine embryos by means of in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT) is limited by great inefficienciy. The present study investigated chromatin and nucleolar dynamics in porcine embryos developed in vivo (IV) and compared this physiological...

  6. Vitrificación de ovocitos bovinos y su uso en el desarrollo partenogenético de embriones Vitrification of bovine oocytes and its use in the parthenogenetic development of embryos

    Directory of Open Access Journals (Sweden)

    J Ruiz

    2010-01-01

    Full Text Available El objetivo de este trabajo fue evaluar el efecto de la vitrificación en la viabilidad de ovocitos activados químicamente para la producción de embriones partenogenéticos bovinos. Ovocitos bovinos aspirados de ovarios obtenidos en el matadero fueron madurados in vitro por 20-22 horas y se distribuyeron en los siguientes grupos. I (n=76: ovocitos vitrificados/descongelados, II (n=119: ovocitos expuestos a crioprotectantes sin vitrificación y III (n=142: ovocitos control. Los ovocitos fueron vitrificados en microgotas sobre un papel de aluminio preenfriado flotando en nitrógeno líquido, utilizando una solución de equilibrio con 4% de etilenglicol y una solución de vitrificación con 35% de etilenglicol 5% de polivinilpirrolidona y 0,4 M de trehalosa. Las microgotas vitrificadas fueron almacenadas en nitrógeno líquido y fueron descongeladas 1-3 días después del almacenamiento. Los tres grupos de ovocitos se activaron partenogenéticamente por exposición de 4 minutos a 5 µM de ionomicina de Ca a temperatura ambiente seguido de una incubación por 5 horas en 6-dimetilaminopurina a 38,5 ºC en una atmósfera húmeda con 5% CO2. Los embriones se cultivaron en medio mSOF durante 8-9 días. Las tasas de ovocitos sobrevivientes fueron 55,1% y 93,7% para ovocitos vitrificados/descongelados (I y expuestos (II respectivamente. Las tasas de segmentación de 55,3%, 72,3% y 74,6%, y de desarrollo hasta blastocistos fueron 7,1%, 17,4% y 21,7% en los grupos I, II y III respectivamente. Estos resultados demuestran que la técnica de vitrificación ha quedado establecida y permite la producción de embriones partenogenéticos bovinos.The aim of this study was to evaluate vitrification effects on the viability of chemically activated oocytes in order to produce parthenogenetic bovine embryos. Bovine oocytes retrieved from ovaries obtained in a slaughterhouse were matured in vitro for 20-22 hours and then assigned to the following groups: I (n=76

  7. Exploratory study of the association of steroid profiles in stimulated ovarian follicular fluid with outcomes of IVF treatment.

    Science.gov (United States)

    Kushnir, Mark M; Naessén, Tord; Wanggren, Kjell; Hreinsson, Julius; Rockwood, Alan L; Meikle, A Wayne; Bergquist, Jonas

    2016-09-01

    Steroid concentrations in stimulated follicular fluid (sFF) samples have been linked to the quality of oocytes used in IVF treatments. Most of the published studies focused on evaluating the association of the IVF outcomes with only a few of the steroids, measured by immunoassays (IA). We performed a treatment outcome, prospective cohort study using stimulated FF sampled from 14 infertile women undergoing IVF treatment; single oocyte was used per IVF cycle. Fourteen endogenous steroids were analyzed in 22 ovarian follicle aspirations, which corresponded to the embryos used in the IVF. Ten oocytes were associated with live birth (LB) and 12 with no pregnancy (NP). Steroids were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. Differences in distribution of concentrations in association with the pregnancy outcome (LB or NP), and receiver operating characteristic (ROC) curves analysis were performed for the entire cohort and for within-women data. The predominant androgen and estrogen in stimulated sFF were androstenedione (A4) and estradiol (E2), respectively. Lower concentrations of pregnenolone (Pr), lower ratios of A4/ dehydroepiandrosterone (DHEA), testosterone (Te)/DHEA, and greater ratios of E2/Te, and estrone/A4 were observed in sFF samples associated with LB. Among the oocytes associated with NP, in four out of 12 samples total concentration of androgens was above the distribution of the concentrations in the oocytes corresponding to the LB group. Observations of the study indicated increased consumption of precursors and increased biosynthesis of estrogens in the follicles associated with LB. Our data suggest that potentially steroid profiles in sFF obtained during oocyte retrieval may serve as biomarkers for selection of the best embryo to transfer after IVF. PMID:26388251

  8. Exploratory study of the association of steroid profiles in stimulated ovarian follicular fluid with outcomes of IVF treatment.

    Science.gov (United States)

    Kushnir, Mark M; Naessén, Tord; Wanggren, Kjell; Hreinsson, Julius; Rockwood, Alan L; Meikle, A Wayne; Bergquist, Jonas

    2016-09-01

    Steroid concentrations in stimulated follicular fluid (sFF) samples have been linked to the quality of oocytes used in IVF treatments. Most of the published studies focused on evaluating the association of the IVF outcomes with only a few of the steroids, measured by immunoassays (IA). We performed a treatment outcome, prospective cohort study using stimulated FF sampled from 14 infertile women undergoing IVF treatment; single oocyte was used per IVF cycle. Fourteen endogenous steroids were analyzed in 22 ovarian follicle aspirations, which corresponded to the embryos used in the IVF. Ten oocytes were associated with live birth (LB) and 12 with no pregnancy (NP). Steroids were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. Differences in distribution of concentrations in association with the pregnancy outcome (LB or NP), and receiver operating characteristic (ROC) curves analysis were performed for the entire cohort and for within-women data. The predominant androgen and estrogen in stimulated sFF were androstenedione (A4) and estradiol (E2), respectively. Lower concentrations of pregnenolone (Pr), lower ratios of A4/ dehydroepiandrosterone (DHEA), testosterone (Te)/DHEA, and greater ratios of E2/Te, and estrone/A4 were observed in sFF samples associated with LB. Among the oocytes associated with NP, in four out of 12 samples total concentration of androgens was above the distribution of the concentrations in the oocytes corresponding to the LB group. Observations of the study indicated increased consumption of precursors and increased biosynthesis of estrogens in the follicles associated with LB. Our data suggest that potentially steroid profiles in sFF obtained during oocyte retrieval may serve as biomarkers for selection of the best embryo to transfer after IVF.

  9. Role of Embryo Glue as a transfer medium in the outcome of fresh non-donor in-vitro fertilization cycles

    Directory of Open Access Journals (Sweden)

    Neeta Singh

    2015-01-01

    Full Text Available Background: EmbryoGlue is a hyaluronan-enriched embryo transfer (ET medium which aids in implantation of embryos, hence, improves pregnancy rates in in-vitro fertilization-ET cycles (IVF-ET. Aim: To evaluate the role of EmbryoGlue in improving implantation and pregnancy rates. Design and Setting: A prospective case-control study conducted at assisted reproductive center of a tertiary care hospital. Method: In 42 women undergoing IVF, embryos were transferred into 50 μL of EmbryoGlue for 10 min prior to transfer inside uterine cavity. In the control group (n = 42, embryos were transferred to conventional blastocyst culture medium. Statistical analysis was performed using SPSS IBM version 19.0. Results: Clinical pregnancy rate in the study group was 7% higher than the control group. The difference, however, was not statistically significant. In addition, no improvement in implantation rates was observed in the study group. However, significant difference (P = 0.04 in clinical pregnancy rate was observed with the EmbryoGlue in patients with previous IVF failure. In the study group, 50% patients (6/12 with previous IVF failure had successful implantation, but in the control group none of the patients (0/11 with previous implantation failure could achieve pregnancy. Conclusion: It is difficult to conclude a favourable role of EmbryoGlue in IVF-ET cycles with a good prognosis. However, in patients with recurrent implantation failure, it may be considered as a useful transfer medium.

  10. Pilot study on molecular quantitation and sequencing of endometrial cytokines gene expression and their effect on the outcome of in vitro fertilization (IVF cycle

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    D. Sabry

    2014-09-01

    Full Text Available Human trophoblast invasion and differentiation are essential for successful pregnancy outcome. The molecular mechanisms, however, are poorly understood. Interleukin (IL-11, a cytokine, regulates endometrial epithelial cell adhesion. Leukemia inhibitory factor (LIF is one of the key cytokines in the embryo implantation regulation. The present study aimed to assess the levels of LIF, IL-11, and IL-11 α receptor gene expression in the endometrium of women undergoing IVF and correlate their levels with the IVF pregnancy outcome. Also, the study aimed to detect any mutation in these three genes among IVF pregnant and non-pregnant women versus control menstrual blood of fertile women. Endometrial tissue biopsies were taken from 15 women undergoing IVF on the day of oocyte retrieval. The quantitative expression of IL-11, IL-11Rα, and LIF genes was assessed by real-time PCR and PCR products were sequenced. Menstrual blood from 10 fertile women was used as control to compare the DNA sequence versus DNA sequence of the studied genes in endometrial biopsies. LH, FSH, and E2 were assessed for enrolled patients by ELISA. Endometrial thickness was also assessed by pelvic ultrasonography. No significant difference was detected between quantitative expression of the three studied genes and pregnancy IVF outcome. Although DNA sequence changes were found in IL-11 and LIF genes of women with negative pregnancy IVF outcome compared to women with positive pregnancy IVF outcome, no DNA sequence changes were detected for IL-11Rα. Other studied parameters (e.g., age, LH, FSH, E2, and endometrial thickness showed no significant differences or correlation of quantitative expression of the three studied involved genes. Data suggested that there were no significant differences between quantitative expression of IL-11, IL-11Rα, and LIF genes and the IVF pregnancy outcome. The present study may reveal that changes in IL-11 and LIF genes sequence may contribute in

  11. Paid to share: IVF patients, eggs and stem cell research.

    Science.gov (United States)

    Roberts, Celia; Throsby, Karen

    2008-01-01

    Following a recent decision by the human fertilisation and embryology authority (HFEA), British women undergoing in vitro fertilisation (IVF) treatment can be 'paid to share' their eggs with stem cell researchers. The HFEA and the clinic proposing the scheme present this as a 'win-win' arrangement benefiting both infertile women and couples and British science. It is also represented as concurrently both 'business as usual' and an exceptional case. Constituting a significant departure from the previous policy and practice of altruistic donation, the scheme has raised significant concerns among clinicians and activists. Here, we ask what questions feminists can bring to these debates without resorting to a position of either refusal or affirmation. Drawing on diverse materials from public debates, as well as social scientific literature on gamete and embryo donation, we undertake a close analysis of the discursive framing and justification of the proposal. We argue that these discourses are characterised by three linked areas of elision and distinction: treatment and research; eggs and embryos; and donation and selling. Our analysis highlights the need for innovative social, ethical and political consideration of egg sharing for stem cell research. PMID:17919794

  12. Patients' Attitudes towards the Surplus Frozen Embryos in China

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    Xuan Jin

    2013-01-01

    Full Text Available Background. Assisted reproductive techniques have been used in China for more than 20 years. This study investigates the attitudes of surplus embryo holders towards embryos storage and donation for medical research. Methods. A total of 363 couples who had completed in vitro fertilization (IVF treatment and had already had biological children but who still had frozen embryos in storage were invited to participate. Interviews were conducted by clinics in a narrative style. Results. Family size was the major reason for participants’ (discontinuation of embryo storage; moreover, the moral status of embryos was an important factor for couples choosing embryo storage, while the storage fee was an important factor for couples choosing embryo disposal. Most couples discontinued the storage of their embryos once their children were older than 3 years. In our study, 58.8% of the couples preferred to dispose of surplus embryos rather than donate them to research, citing a lack of information and distrust in science as significant reasons for their decision. Conclusions. Interviews regarding frozen embryos, including patients’ expectations for embryo storage and information to assist them with decisions regarding embryo disposal, are beneficial for policies addressing embryo disposition and embryo donation in China.

  13. Derivation of Two New Human Embryonic Stem Cell Lines from Nonviable Human Embryos

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    Svetlana Gavrilov

    2011-01-01

    Full Text Available We report the derivation and characterization of two new human embryonic stem cells (hESC lines (CU1 and CU2 from embryos with an irreversible loss of integrated organismic function. In addition, we analyzed retrospective data of morphological progression from embryonic day (ED 5 to ED6 for 2480 embryos not suitable for clinical use to assess grading criteria indicative of loss of viability on ED5. Our analysis indicated that a large proportion of in vitro fertilization (IVF embryos not suitable for clinical use could be used for hESC derivation. Based on these combined findings, we propose that criteria commonly used in IVF clinics to determine optimal embryos for uterine transfer can be employed to predict the potential for hESC derivation from poor quality embryos without the destruction of vital human embryos.

  14. Efeito de diferentes meios de cultivo no desenvolvimento e proporção do sexo de embriões bovinos produzidos in vitro Effect of different culture media on development and sex ratio of bovine embryos fertilized in vitro

    Directory of Open Access Journals (Sweden)

    S.G.T. Gilardi

    2004-10-01

    Full Text Available Avaliou-se o efeito da suplementação de meios de cultivo sobre o desenvolvimento e proporção do sexo de embriões bovinos fertilizados in vitro. Complexos cumulus-oócitos obtidos de ovários de matadouro foram maturados e fertilizados in vitro. Os zigotos (n= 484 foram distribuídos aleatoriamente em meio CR2aa, contendo soro fetal bovino (SFB (T1, albumina sérica bovina (BSA (T2 ou BSA mais insulina:transferrina:selênio e vitaminas (BSA+ (T3, no cultivo embrionário in vitro, a uma atmosfera de 5% CO2 a 38,8ºC em ar. A taxa de clivagem foi observada 72-76 horas pós-fertilização (PF e a taxa de blastocistos com sete e oito dias PF. Os blastocistos (n= 63 foram sexados pela técnica de reação em cadeia de polimerase. A taxa de clivagem em T2 foi maior (P0,05 entre T2 e T3, porém menor (P0,05 entre os tratamentos. O T1 influenciou o desenvolvimento de blastocistos, mas não teve efeito sobre a proporção do sexo.The effect of culture media on the development and on the sex ratio of bovine embryos fertilized in vitro was studied. Cumulus oocyte-complexes from slaughterhouse ovaries were matured and fertilized in vitro. Zygotes (n= 484 were randomly allotted to different culture media and cultured with their cumulus cells in CR2aa medium and an atmosphere of 5% CO2 in air at 38.8ºC. The fetal calf serum (FCS, bovine seric albumin (BSA or BSA plus insulin:transferrin:selenium and vitamins (BSA+ supplementation effect on embryo culture was evaluated. Cleavage rate was assessed at 72-76h post-fertilization (PF and blastocyst rate on days 7 and 8 PF. The blastocysts (n= 63 were also sexed using polymerase chain reaction. Cleavage rate for BSA medium supplemented was higher (P0.05, but lower (P<0.01 than FCS. Culture medium FCS supplemented affected blastocyst development but not the sex ratio.

  15. 性控精液在牛体外性控胚胎生产中的应用%Application of Sexed Semen on the in vitro Bovine Sexed Embryos

    Institute of Scientific and Technical Information of China (English)

    孙凤俊; 张佳谊; 韩广文

    2013-01-01

    Use of sexed semen in conjunction with in vitro embryo production is a potentially efficient means of obtaining offspring of predetermined sex. The ability to sort individual sperm cells into viable X- and Y-chromosome-bearing fractions made producers' sex selection dreams reality in the 1990s and now semen can be sexed with greater than 90% accuracy with use of a flow cytometric cell sorter. However, there are some drawbacks to implementing the extensive use of sexed semen technology include the apparent lower fertility of sorted sperm, the lower survival of sorted sperm after cryopreservation and the reduced number of sperm that could be separated in a specified time period. The issues of In vitro production of bovine embryos using sexed semen and its application prospect are discussed in this review.%利用性控精液结合体外胚胎生产技术是获得预知性别后代的一种有潜在效率的方法.自20世纪90年代,利用流式细胞仪将X精子和Y精子分离开来,使畜牧生产性别选择的梦想变为现实,性控精液得到了极大的推广应用.本文重点阐述利用性控精液进行胚胎体外生产的研究和生产状况.

  16. Emotions and Ethical Considerations of Women Undergoing IVF-Treatments

    NARCIS (Netherlands)

    Kaliarnta, S.; Nihlén-Fahlquist, J.; Roeser, S.

    2011-01-01

    Women who suffer from fertility issues often use in vitro fertilization (IVF) to realize their wish to have children. However, IVF has its own set of strict administration rules that leave the women physically and emotionally exhausted. Feeling alienated and frustrated, many IVF users turn to intern

  17. In vitro fertilization in Japan — Early days of in vitro fertilization and embryo transfer and future prospects for assisted reproductive technology —

    OpenAIRE

    SUZUKI, Masakuni

    2014-01-01

    Assisted reproductive technology (ART) such as in vitro fertilization (IVF) and embryo transfer (ET) has been essential in the treatment of infertility. The world’s first IVF-ET baby was born in 1978 based on the technique developed by Dr. Robert Edwards and Dr. Patrick Steptoe. 1) In Japan, the first IVF-ET birth was reported in 1983 by Prof. Masakuni Suzuki at Tohoku University School of Medicine. 2,3) IVF-ET is a procedure used to achieve pregnancy that consists of extracting oocytes from ...

  18. Regimen of ovarian stimulation affects oocyte and therefore embryo quality.

    Science.gov (United States)

    Bosch, Ernesto; Labarta, Elena; Kolibianakis, Efstratios; Rosen, Mitchell; Meldrum, David

    2016-03-01

    Without any doubt the regimen used to mature multiple capable oocytes for IVF impacts IVF outcomes. Studies have indicated that the inclusion of LH activity, adjuvant agents such as growth hormone (GH), and regimens providing for simultaneous action of both LH and FSH during final oocyte maturation may have beneficial effects on IVF outcomes. Because of the difficulty in improving IVF outcomes in poor responders, the studies on GH are of particular interest. As pointed out in this review, the apparent beneficial effects of GH on oocyte competence may also apply to older women or to normal responders with reduced embryo quality. A much more difficult question is whether and how much ovarian stimulation impacts on oocyte competence. Paradoxically it seems that there are not demonstrated differences between the stimulated and the natural unstimulated cycle, whereas studies in laboratory animals and IVF patients have shown deleterious effects of higher compared with lower doses of gonadotropins. Recent studies suggest that the use of high doses of gonadotropins as an independent factor correlates negatively with the probability of live birth, whereas a high ovarian response per se is associated with better cumulative pregnancy rates, owing to the availability of more euploid and good-quality embryos. Although adjunctive use of androgens has not been discussed here, it is briefly covered in the first review of this series. PMID:26826273

  19. Analysis of ovarian reserve markers (AMH, FSH, AFC) in different age strata in IVF/ICSI patients

    Science.gov (United States)

    Shahrokh Tehraninezhad, Ensieh; Mehrabi, Fatemeh; Taati, Raheleh; Kalantar, Vahid; Aziminekoo, Elham; Tarafdari, Azam

    2016-01-01

    Background: The predictive roles of follicle stimulating hormone (FSH), anti-mullerian hormone (AMH) and antral follicle count (AFC) as ovarian reserve markers in women with different age groups are not established well. Objective: This study compares the value of FSH, AMH and AFC at the time of in vitro fertilization (IVF) treatment in different age groups. Materials and Methods: In this cross-sectional study, 103 women aged 20-43 years candidates for IVF/ICSI cycle were recruited. FSH, AMH and AFC on day 3 of menstrual cycle were measured. The relationship of these measured markers with outcome variables (oocytes number, number of frozen/fresh embryo and chemical and clinical pregnancy) was assessed in different age groups (i.e. 20-32, 33-37 and 38-43 years). Results: our results show that age was correlated with clinical pregnancy, oocyte count and fresh and frozen embryo (p<0.001). AMH, AFC and FSH were not correlated with clinical or chemical pregnancy at total population or age subgroups except the significant correlation of AFC with clinical pregnancy at 33-37 years old group. AFC was correlated with oocyte count and the number of fresh and frozen embryos in the ages group 20-32 years. In this age group, AMH was correlated with fresh and frozen embryos. AMH, AFC and FSH were correlated with oocyte count and the number of fresh embryos in age group 33-37 years. AMH was correlated with oocyte count and the number of fresh embryos in 38-43 years old group. Conclusion: We concluded that the age is the superior predictor of IVF outcome and AMH and AFC are variable predicting markers of ovarian reserve in different age groups. PMID:27679824

  20. 体外受精-胚胎移植技术的建立及其衍生技术的发展%The establishment of in vitro fertilization-embryo transfer (IVF-ET) and the development of its de-rivative techniques

    Institute of Scientific and Technical Information of China (English)

    尚欣妹; 李志平

    2009-01-01

    1978年7月25日首例运用体外受精-胚胎移植技术(IVF-ET)而分娩的试管婴儿诞生,它为人类助孕技术的发展翻开新的篇章.此后,相继出现如:经腹腔精子与卵子移植技术(POST)、配子输卵管内移植技术(GIFT)、单精子卵细胞胞浆内注射(ICSI)、着床前遗传诊断技术(PDG)等衍生技术.这些技术的出现对体外受精-胚胎移植技术进行全面补充,使助孕技术更趋于完善.通过对此项技术及其衍生技术的回顾,会使人们更全面的了解此类技术并为其更健康地发展提供经验和启迪.%On 25th July,1978, a test-tube baby from the first use of IVF-ET was born, which opened a new chapter for the development of ART for human beings. Thereafter, some derivative techniques such as POST, GIFT, ICSI and PDG, etc., emerged in succession, which were a comprehensive supple-ment to IVF-ET, and further perfected ART. With the review of this technique and its derivative tech-niques, people can more comprehensively understand the techniques and provide experience and enlighten-ment for its more healthy development.

  1. Cumulative pregnancy rates after sequential treatment with modified natural cycle IVF followed by IVF with controlled ovarian stimulation

    NARCIS (Netherlands)

    Pelinck, M. J.; Knol, H. M.; Vogel, N. E. A.; Arts, E. G. J. M.; Simons, A. H. M.; Heineman, M. J.; Hoek, A.

    2008-01-01

    BACKGROUND: In modified natural cycle IVF (MNC-IVF), treatment is aimed at using the one follicle that spontaneously develops to dominance, using a GnRH-antagonist together with gonadotrophins in the late follicular phase only. The MNC-IVF is of interest because of its low-risk and patient-friendly

  2. Is the birthweight of singletons born after IVF reduced by ovarian stimulation or by IVF laboratory procedures?

    NARCIS (Netherlands)

    Pelinck, M. J.; Hadders-Algra, M.; Haadsma, M. L.; Nijhuis, W. L.; Kiewiet, S. M.; Hoek, A.; Heineman, M. J.; Middleburg, K. J.

    2010-01-01

    Singletons born after IVF are at risk of adverse pregnancy outcome, the cause of which is unknown. The present study investigated the influence of ovarian stimulation and IVF laboratory procedure on birthweight. Birthweight of singleton pregnancies resulting from IVF treatment with (n = 161) and wit

  3. Effect of granulocyte colony stimulating factor (G-CSF) on IVF outcomes in infertile women: An RCT

    Science.gov (United States)

    Eftekhar, Maryam; Hosseinisadat, Robabe; Baradaran, Ramesh; Naghshineh, Elham

    2016-01-01

    Background: Despite major advances in assisted reproductive techniques, the implantation rates remain relatively low. Some studies have demonstrated that intrauterine infusion of granulocyte colony stimulating factor (G-CSF) improves implantation in infertile women. Objective: To assess the G-CSF effects on IVF outcomes in women with normal endometrial thickness. Materials and methods: In this randomized controlled clinical trial, 100 infertile women with normal endometrial thickness who were candidate for IVF were evaluated in two groups. Exclusion criteria were positive history of repeated implantation failure (RIF), endocrine disorders, severe endometriosis, congenital or acquired uterine anomaly and contraindication for G-CSF (renal disease, sickle cell disease, or malignancy). In G-CSF group (n=50), 300 µg trans cervical intrauterine of G-CSF was administered at the oocyte retrieval day. Controls (n=50) were treated with standard protocol. Chemical, clinical and ongoing pregnancy rates, implantation rate, and miscarriage rate were compared between groups. Results: Number of total and mature oocytes (MII), two pronuclei (2PN), total embryos, transferred embryos, quality of transferred embryos, and fertilization rate did not differ significantly between two groups. So there were no significant differences between groups in chemical, clinical and ongoing pregnancy rate, implantation rate, and miscarriage rate Conclusion: our result showed in normal IVF patients with normal endometrial thickness, the intrauterine infusion of G-CSF did not improve pregnancy outcomes. PMID:27326420

  4. Does the use of gonadotropin-releasing hormone antagonists in natural IVF cycles for poor responder patients cause more harm than benefit?

    Science.gov (United States)

    Aksoy, Senai; Yakin, Kayhan; Seyhan, Ayse; Oktem, Ozgur; Alatas, Cengiz; Ata, Baris; Urman, Bulent

    2016-06-01

    Poor ovarian response to controlled ovarian stimulation (COS) is one of the most critical factors that substantially limits the success of assisted reproduction techniques (ARTs). Natural and modified natural cycle IVF are two options that could be considered as a last resort. Blocking gonadotropin-releasing hormone (GnRH) actions in the endometrium via GnRH receptor antagonism may have a negative impact on endometrial receptivity. We analysed IVF outcomes in 142 natural (n = 30) or modified natural (n = 112) IVF cycles performed in 82 women retrospectively. A significantly lower proportion of natural cycles reached follicular aspiration compared to modified natural cycles (56.7% vs. 85.7%, p numbers of IVF cycles ending in embryo transfer (26.7% vs. 44.6%) was not statistically significant between natural cycle and modified natural IVF cycles. Clinical pregnancy (6.7% vs. 7.1%) and live birth rates per initiated cycle (6.7% vs. 5.4%) were similar between the two groups. Notably, the implantation rate was slightly lower in modified natural cycles (16% vs. 25%, p > 0.05). There was a trend towards higher clinical pregnancy (25% vs. 16%) and live birth (25% vs. 12%) rates per embryo transfer in natural cycles compared to modified natural cycles, but the differences did not reach statistical significance.

  5. Embryo apoptosis identification: Oocyte grade or cleavage stage?

    Science.gov (United States)

    Bakri, Noraina Mohd; Ibrahim, Siti Fatimah; Osman, Nurul Atikah; Hasan, Nurhaslina; Jaffar, Farah Hanan Fathihah; Rahman, Zulaiha Abdul; Osman, Khairul

    2016-01-01

    Apoptosis is a programed cell death that is vital for tissue homeostasis. However, embryo apoptosis had been known to be related to embryo fragmentation which should be avoided in in vitro fertilization (IVF). The purpose of this study was to evaluate the relationship of embryo apoptosis with the grade of immature oocytes and cleavage stage of in vitro produced (IVP) cattle embryos. This study consisted of 345 oocytes collected through ovary slicing. Immature oocytes were graded as A, B and C. This grading was based on cumulus cell thickness and compactness. All oocytes then underwent an in vitro maturation (IVM) procedure. An IVF was done 24 h after IVM culture. Prior to staining, stage of cleaved embryos was determined and classified as either 2, 4, 8 or >8-cell embryo stage. Apoptosis status of cleaved IVP embryos was determined by using annexin V-FITC staining technique at 48 and 72 h post insemination (hpi). Apoptosis status for each embryo was classified as either early or late. The result showed that there was no significant difference (p > 0.05) of apoptosis status among grade A, B and C embryos. All grades of oocytes showed embryo apoptosis where 1.5% late apoptosis for grade A, 4.5% and 10.4% of early and late apoptosis for grade B and grade C. Early apoptosis was not seen in grade A embryo. We also noted no significant difference (p > 0.05) of apoptosis status between 2, 4, 8 and >8-cell embryo stage. Early apoptosis was also not seen in >8-cell stage. Even though there were no differences in apoptosis expression between the three classes, the cleavage rate of grade A oocytes was significantly higher (p quality and the cleavage stage of the embryo produced. PMID:26858565

  6. Pronuclear morphology evaluation in in vitro fertilization (IVF / intracytoplasmic sperm injection (ICSI cycles: a retrospective clinical review

    Directory of Open Access Journals (Sweden)

    Nicoli Alessia

    2013-01-01

    Full Text Available Abstract Background The assessment of the embryo quality is crucial to maintain an high pregnancy rate and to reduce the risk of multiple pregnancy. The evaluation of the pronuclear and nucleolar characteristics of human zygote have been proposed as an indicator of embryo development and chromosomal complement. The aim of the current study was to assess the role of pronuclear morphology evaluation in vitro fertilization (IVF / intracytoplasmic sperm injection (ICSI cycles. Methods Retrospective clinical analysis on 755 non-elective transfers of only one embryo (ET. Embryo assessment was performed in days 1 and 2. Clinical and biological data were recorded and analyzed according to embryo and/or pronuclear morphology. Results Both pronuclear and embryo morphology were significantly related to clinical pregnancy and live-birth rates. No significant difference in clinical pregnancy and live-birth rates was detected when the pronuclear and embryo morphology assessments were combined. Embryo morphology and maternal age were the only independent predictors of favorable outcome by logistic regression analysis. Conclusions Pronuclear evaluation is effective to select the best zygotes if ET is performed at day 1, whereas it did not improve the clinical outcomes when combined with embryo morphology evaluation in day 2.

  7. Assisted conception- IVF and other procedures

    OpenAIRE

    Brincat, Mark; Duca, Edward

    2013-01-01

    Assisted conception procedures arose as a type of treatment for infertility. They opened a whole range of possibilities for couples that were unable to have children due to a variety of problems. http://www.um.edu.mt/think/assisted-conception-ivf-and-other-procedures/

  8. The Psychological Impact of IVF Treatment

    NARCIS (Netherlands)

    C. de Klerk (Cora)

    2008-01-01

    textabstractIVF treatment requires the woman to undergo several invasive procedures, which are repeated in subsequent treatment cycles. In addition to the physical burden, the threat of treatment failure confronts the couple with the possibility that they have to give up hope to have a child of thei

  9. Milder ovarian stimulation for in-vitro fertilization reduces aneuploidy in the human preimplantation embryo : a randomized controlled trial

    NARCIS (Netherlands)

    Baart, Esther B.; Martini, Elena; Eijkemans, Marinus J.; Van Opstal, Diane; Beckers, Nicole G. M.; Verhoeff, Arie; Macklon, Nicolas S.; Fauser, Bart C. J. M.

    2007-01-01

    To test whether ovarian stimulation for in-vitro fertilization (IVF) affects oocyte quality and thus chromosome segregation behaviour during meiosis and early embryo development, preimplantation genetic screening of embryos was employed in a prospective, randomized controlled trial, comparing two ov

  10. Derivation and characterisation of hESC lines from supernumerary embryos, experience from Odense, Denmark

    DEFF Research Database (Denmark)

    Harkness, Linda; Rasmussen, Iben Anne; Erb, Karin;

    2010-01-01

    The derivation and characterisation of human embryonic stem cells provides a source of pluripotent stem cells with potential for clinical applications. Utilising locally sourced embryos from two IVF clinics, we derived and characterised five new cell lines for use in a non-clinical setting....... Analysis of clinical data showed that the majority of embryos (94.5%) failed to reach the blastocyst stage of development and of all embryos, regardless of developmental status, 248 embryos were needed to create one stem cell line. From the number of embryos (69) which developed to the blastocyst stage 8...... available through the UK Stem Cell Bank ( http://www.ukstemcellbank.org.uk/ )....

  11. Steroid hormones in bovine oviductal fluid during the estrous cycle.

    Science.gov (United States)

    Lamy, Julie; Liere, Philippe; Pianos, Antoine; Aprahamian, Fanny; Mermillod, Pascal; Saint-Dizier, Marie

    2016-10-01

    Ovarian steroid hormones are major regulators of the physiology of the oviduct and reproductive events occurring within the oviduct. To establish a whole steroid profiling of the bovine oviductal fluid (OF) during the estrous cycle, contralateral and ipsilateral (to the corpus luteum or preovulatory follicle) oviducts were classified into four stages of the estrous cycle (n = 18-27 cows per stage): postovulatory (Post-ov), mid-luteal (Mid-lut), late luteal (Late-lut), and preovulatory on the basis of the ovarian morphology and intrafollicular steroid concentrations. Steroids were extracted from pools of 150 to 200 μL OF (three to 10 cows per pool; three to four pools per "stage × side" group), purified, fractioned by high-performance liquid chromatography, and analyzed by gas chromatography coupled with tandem mass spectrometry. The concentrations of progesterone (P4) in ipsilateral OF increased from Post-ov (56.9 ± 13.4 ng/mL) to Mid-lut (120.3 ± 34.3 ng/mL), then decreased from Late-lut (76.7 ± 1.8 ng/mL) to Pre-ov (6.3 ± 1.7 ng/mL), and were four to 16 times higher than in contralateral OF. Most P4 metabolites followed similar patterns of variation. Concentrations of 17beta-estradiol (E2) were significantly higher at Pre-ov (290.5 ± 63.2 pg/mL) compared with all other stages (environment was evidenced in the bovine OF. These results could be useful to improve media for IVF, embryo development, and culture of oviductal cells. PMID:27262884

  12. Resistin levels of serum and follicular fluid in non-obese patients with polycystic ovary syndrome during IVF cycles

    Institute of Scientific and Technical Information of China (English)

    LU Xiu-e; HUANG He-feng; LI Mei-gen; ZHU Yi-min; QIANG Yu-li; DONG Min-yue

    2005-01-01

    Objectives: To measure serum and follicular resistin, steroids hormone levels in women with PCOS (polycystic ovary syndrome) (BMI (body mass index)<25 kg/m2), to assess possible correlations of resistin to hormonal and metabolic parameters and to analyze the clinical outcomes of in vitro fertilization-embryo transfer (IVF-ET) in women with PCOS and tubal infertility. Study design: We analyzed the clinical outcomes of IVF-ET in women with PCOS (BMI<25 kg/m2) and tubal infertility during the years 2002 to 2004 and compared the serum and follicular fluid resistin levels, estradiol (E2), progesterone (P), testosterone (T)levels in 20 PCOS and 20 healthy, age-matched women without PCOS during IVF-stimulated cycles. The correlations between the resistin levels and the outcomes of IVF-ET were evaluated. Results: No significant differences in resistin levels of either serum or follicular fluid between PCOS and control group were found. However, resistin levels in serum were higher than that in follicular fluid in both groups. Multiple regression analysis showed that resistin levels in serum did not correlate with BMI, estradiol, LH (luteinizing hormone) and insulin level in fasting blood. No significant correlations were found between follicular fluid reisistin levels and fertilization rate, implantation rate, clinical pregnancy rate or early miscarriage rate in both PCOS and control groups.Conclusion: Our results show that resistin does not have correlation with the hormonal and metabolic parameters as well as the outcomes of IVF. These data suggest that resistin is unlikely to be a local determinant factor in steroidogenesis and growth and maturation of oocytes during IVF-ET in lean women with PCOS.

  13. Couples with unexplained subfertility and unfavorable prognosis: a randomized pilot trial comparing the effectiveness of in vitro fertilization with elective single embryo transfer versus intrauterine insemination with controlled ovarian stimulation

    NARCIS (Netherlands)

    I.M. Custers; T.E. König; F.J. Broekmans; P.G.A. Hompes; E. Kaaijk; J. Oosterhuis; M.H. Mochtar; S. Repping; M. van Wely; P. Steures; F. van der Veen; B.W.J. Mol

    2011-01-01

    To evaluate the effectiveness of IVF with elective single embryo transfer (IVF-eSET) vs. IUI with controlled ovarian stimulation (IUI-COS) as an alternative treatment to reduce the risk for a multiple pregnancy. Randomized pilot trial. Three academic and six teaching hospitals in the Netherlands. Co

  14. Couples with unexplained subfertility and unfavorable prognosis : a randomized pilot trial comparing the effectiveness of in vitro fertilization with elective single embryo transfer versus intrauterine insemination with controlled ovarian stimulation

    NARCIS (Netherlands)

    Custers, Inge M.; Konig, Tamar E.; Broekmans, Frank J.; Hompes, Peter G. A.; Kaaijk, Eugenie; Oosterhuis, Jur; Mochtar, Monique H.; Repping, Sjoerd; van Wely, Madelon; Steures, Pieternel; van der Veen, Fulco; Mol, Ben W. J.

    2011-01-01

    Objective: To evaluate the effectiveness of IVF with elective single embryo transfer (IVF-eSET) vs. IUI with controlled ovarian stimulation (IUI-COS) as an alternative treatment to reduce the risk for a multiple pregnancy. Design: Randomized pilot trial. Setting: Three academic and six teaching hosp

  15. EFFECT OF PELVIC ENDOMETRIOSIS, ENDOMETRIOMAS AND RECURRENT ENDOMETRIOMAS ON IVF-ET/ICSI OUTCOMES

    Science.gov (United States)

    Xing, Weijie; Lin, Haiyan; Wu, Zexuan; Li, Yu; Zhang, Qingxue

    2016-01-01

    Introduction: Endometriosis, the most common gynecological disorder, is a challenging disease observed in 20% - 40% of subfertile women. Material and Methods: 380 women were divided into four groups. Group A consisted of 176 women with pelvic endometriosis. Group B consisted of 125 women who had previously undergone a laparoscopic endometrioma cystectomy. Group C consisted of 38 women with recurrent endometriomas without aspiration before IVF-ET/ICSI. Group D consisted of 41 women with recurrent endometriomas undergone aspiration before IVF-ET/ICSI. Results: Baseline FSH level (8.61 ± 3.42 mIU/mL) and total dose of Gn (2337.15 ± 853.00 IU) in Group A were the lowest (p < 0.05). The number of retrieved oocytes in Group B (7.98 ± 5.05) was significantly fewer than those in Group A and D (p < 0.05). The numbers of MII oocytes in Groups A, C and D were significantly larger than that in Group B. The number of retrieved oocytes, high-quality embryos, implantation and pregnancy rates were similar in Groups C and D. Conclusions: Pelvic endometriosis had a less adverse effect on ovarian reserve than endometrioma. No advantage was found in transvaginal aspiration for recurrent endometriomas before IVF-ET/ICSI. PMID:27147911

  16. 体外受精-胚胎移植中冷冻胚胎处置的伦理思考(附34例分析)%Ethical consideration of the disposal of frozen embryos in IVF.(analysis of 34 cases)

    Institute of Scientific and Technical Information of China (English)

    张静雯; 马文敏; 阳星娥; 黄绮云; 黄建洲; 池霖生

    2013-01-01

    目的 探讨辅助生殖技术施行过程中产生的冷冻胚胎处置的伦理问题.方法 回顾分析2010年4月至2013年1月期间进行冷冻胚胎销毁的34个实例,项目包括胚胎销毁的数量、进行胚胎销毁的原因、知情谈话的结果等.结果 共有41对夫妇要求进行胚胎销毁处理,经过知情谈话,其中7对夫妇暂时取消胚胎销毁决定,34对夫妇坚持进行胚胎销毁处理.其中共销毁胚胎262个,胚胎销毁原因中,离婚占11.8%,已生育健康小孩占70.6%,转诊治疗占2.9%,对治疗绝望要求放弃治疗5.9%,胚胎解冻后夫妇意见不统一5.9%,不明原因2.9%.结论 患者要求对冷冻胚胎进行销毁的权利不容忽视,生殖中心无权违背患者夫妇双方的意愿进行胚胎保存,冷冻胚胎处置已经成为生殖领域亟须解决的问题,应进一步研究冷冻胚胎处置的伦理问题,完善冷冻胚胎处置的操作流程.%Objective:To explore the ethical issues of assisted reproductive technology implementation process of disposal of frozen embryos.Methods:A retrospective analysis of frozen embryos destroyed during the 34 instances from April 2010 to January 2013,including the number of embryo destruction,the destruction of embryos reason,informed of the results:of the conversation.Results:A total of 41 couples require embryo destruction,after informed conversation,of which seven couples decided to suspended embryos destroyed,34 couples adhere to the embryo destroyed.Which have destroyed 262 embryos,embryos Destruction reason,divorce accounted for 11.8%,70.6%,has given birth to healthy children,accounted for 2.9% of referral for treatment,the treatment of despair asked to give up treatment 5.9%,thawed embryos couples disagreement 5.9% 2.9% of the unexplained.Conclusion:Patients the right to require the destruction of frozen embryos can not be ignored,the Center for Reproductive no right to go against the wishes of the patients with couples

  17. High Serum FSH is Associated with Brown Oocyte Formation and a Lower Pregnacy Rate in Human IVF Parctice

    Directory of Open Access Journals (Sweden)

    Hongyi Xu

    2016-07-01

    Full Text Available Background/Aims: To investigate whether brown zona pellucida (ZP of oocytes affects the outcome of fertilization, embryo quality and pregnancy rate in in vitro fertilization-embryo transfer (IVF-ET. Methods: Based on the ZP color of their oocytes, a total number of 703 patients dated from 2012 to 2014 were divided into a normal egg group (group A and a brown oocyte group (group B, with 629 and 74 cases, respectively. Clinical characteristics, gonadotropin (Gn days, Gn dosage, serum hormone levels on the day of human chorionic gonadotropin (HCG injection, ZP thickness (ZPT of the eggs, fertilization rate, rescue intracytoplasmic sperm injection (rICSI rate, good-quality embryo rate and pregnancy rate were compared between the two groups. Results: No significant differences were found in the duration and the causes of infertility, and their basal level of endocrine hormone before IVF-ET between normal egg group and brown egg group. The level of serum hormone including estradiol, progesterone and luteinizing hormone on the day of HCG injection were again similar. Moreover, there were no differences in number of mature oocytes, oocyte fertilization rates and rICSI rates after IVF between the two groups. However, we observed that the ZPT of brown oocytes (group B was higher than that of normal oocytes (group A. Moreover, the Gn dosage and FSH levels on the day of HCG injection were significantly higher in group B than in group A and the good-quality embryo rate and pregnancy rate in group B were lower than those in group A. Conclusion: Compared with normal eggs, oocytes with a brown ZP were found to have a higher ZPT, lower embryo quality and lower pregnancy rate, which might be due to a high Gn dosage injection and high serum FSH levels during IVT-ET cycles.

  18. The INeS study: prevention of multiple pregnancies: a randomised controlled trial comparing IUI COH versus IVF e SET versus MNC IVF in couples with unexplained or mild male subfertility

    Directory of Open Access Journals (Sweden)

    Beckers Nicole

    2009-12-01

    Full Text Available Abstract Background Multiple pregnancies are high risk pregnancies with higher chances of maternal and neonatal mortality and morbidity. In the past decades the number of multiple pregnancies has increased. This trend is partly due to the fact that women start family planning at an increased age, but also due to the increased use of ART. Couples with unexplained or mild male subfertility generally receive intrauterine insemination IUI with controlled hormonal stimulation (IUI COH. The cumulative pregnancy rate is 40%, with a 10% multiple pregnancy rate. This study aims to reveal whether alternative treatments such as IVF elective Single Embryo Transfer (IVF e SET or Modified Natural Cycle IVF (MNC IVF can reduce the number of multiple pregnancy rates, but uphold similar pregnancy rates as IUI COH in couples with mild male or unexplained subfertility. Secondly, the aim is to perform a cost effective analyses and assess treatment preference of these couples. Methods/Design We plan a multicentre randomised controlled clinical trial in the Netherlands comparing six cycles of intra-uterine insemination with controlled ovarian hyperstimulation or six cycles of Modified Natural Cycle (MNC IVF or three cycles with IVF-elective Single Embryo Transfer (eSET plus cryo-cycles within a time frame of 12 months. Couples with unexplained subfertility or mild male subfertility and a poor prognosis for treatment independent pregnancy will be included. Women with anovulatory cycles, severe endometriosis, double sided tubal pathology or serious endocrine illness will be excluded. Our primary outcome is the birth of a healthy singleton. Secondary outcomes are multiple pregnancy, treatment costs, and patient experiences in each treatment arm. The analysis will be performed according tot the intention to treat principle. We will test for non-inferiority of the three arms with respect to live birth. As we accept a 12.5% loss in pregnancy rate in one of the two IVF arms

  19. Study on HBV Vertical Transmission via the in vitro Fertilization(IVF)Technique

    Institute of Scientific and Technical Information of China (English)

    Jing-ning YANG; Qing-bin LUO; Chang-jun ZHANG; Hua WANG

    2009-01-01

    Objective To study the hepatitis B virus(HB V)vertical transmission via infected spermatozoa.Methods Eighteen male patients with HBV infection who underwent in vitro fertilization (IVF)were studied,5 HBV negative patients were selected as the control.Fluorescence in situ hybridization(FISH)analysis using the partial-length HBV DNA as the hybridization probe was performed to explore the existence of HBV DNA in the sperm and in the host embryonic genome.Results FISH showed that 5 of 18 patients' sperm presented positive signals and 2 of 18 embryos presented positive signals,while no positive signals were found in control group.Conclusion The HBV DNA was found in human sperm and embryos of HBV patients.These results provide direct evidence that HBV DNA could transmit to foetus via human infected spermatozoa.

  20. Effect of Embryo Transfer on the Bovine Estrus Synchronization and Conception%胚胎移植受体牛同期发情及受胎效果的研究

    Institute of Scientific and Technical Information of China (English)

    李刚

    2012-01-01

    The method of CIDR+E2 and twice use of uterine PGF2α infusion method were used in this study, embryo transfer for cattle estrus synchronization and fresh embryo transfer pregnancy was observed. The results showed that receptor bovine estrous rate at 24--48h was 85.5% and 51.6%, there was significantly difference between them (P〈0.05). Cattle before transplantation luteal receptor inspection pass rate was 79.3%, 77.8%, there was no significant difference (P〉0.05). Conception rate was 45.8% and 42.4%, the difference was not significant (P〉0.05). Estrus observed after 24h, 6.5d for direct examination Cattle with luteinizing for A, B grade can be used for transplantation.%本试验分别采用CIDR+E2法和两次PGF2α子宫输注法,对胚胎移植受体牛做同期发情处理,鲜胚移植观察其妊娠情况。试验结果表明两种处理方法的受体牛在24-48h发情率分别为85.5%和51.6%,两者之间差异显著(P〈0.05)。移植前对受体牛进行黄体检查,黄体合格率分另11为79.3%和77.8%,差异不显著(P〉0.05);受胎率分别为45.8%和42.4%,差异不显著(P〉0.05)。24h后观察发情,6.5d直检,黄体A、B级者用于移植。

  1. Embryo aggregation does not improve the development of interspecies somatic cell nuclear transfer embryos in the horse.

    Science.gov (United States)

    Gambini, Andrés; De Stéfano, Adrián; Jarazo, Javier; Buemo, Carla; Karlanian, Florencia; Salamone, Daniel Felipe

    2016-09-01

    The low efficiency of interspecies somatic cell nuclear transfer (iSCNT) makes it necessary to investigate new strategies to improve embryonic developmental competence. Embryo aggregation has been successfully applied to improve cloning efficiency in mammals, but it remains unclear whether it could also be beneficial for iSCNT. In this study, we first compared the effect of embryo aggregation over in vitro development and blastocyst quality of porcine, bovine, and feline zona-free (ZF) parthenogenetic (PA) embryos to test the effects of embryo aggregation on species that were later used as enucleated oocytes donors in our iSCNT study. We then assessed whether embryo aggregation could improve the in vitro development of ZF equine iSCNT embryos after reconstruction with porcine, bovine, and feline ooplasm. Bovine- and porcine-aggregated PA blastocysts had significantly larger diameters compared with nonaggregated embryos. On the other hand, feline- and bovine-aggregated PA embryos had higher blastocyst cell number. Embryo aggregation of equine-equine SCNT was found to be beneficial for embryo development as we have previously reported, but the aggregation of three ZF reconstructed embryos did not improve embryo developmental rates on iSCNT. In vitro embryo development of nonaggregated iSCNT was predominantly arrested around the stage when transcriptional activation of the embryonic genome is reported to start on the embryo of the donor species. Nevertheless, independent of embryo aggregation, equine blastocyst-like structures could be obtained in our study using domestic feline-enucleated oocytes. Taken together, these results reported that embryo aggregation enhance in vitro PA embryo development and embryo quality but effects vary depending on the species. Embryo aggregation also improves, as expected, the in vitro embryo development of equine-equine SCNT embryos; however, we did not observe positive effects on equine iSCNT embryo development. Among oocytes

  2. Digital Microfluidic Dynamic Culture of Mammalian Embryos on an Electrowetting on Dielectric (EWOD) Chip.

    Science.gov (United States)

    Huang, Hong-Yuan; Shen, Hsien-Hua; Tien, Chang-Hung; Li, Chin-Jung; Fan, Shih-Kang; Liu, Cheng-Hsien; Hsu, Wen-Syang; Yao, Da-Jeng

    2015-01-01

    Current human fertilization in vitro (IVF) bypasses the female oviduct and manually inseminates, fertilizes and cultivates embryos in a static microdrop containing appropriate chemical compounds. A microfluidic microchannel system for IVF is considered to provide an improved in-vivo-mimicking environment to enhance the development in a culture system for an embryo before implantation. We demonstrate a novel digitalized microfluidic device powered with electrowetting on a dielectric (EWOD) to culture an embryo in vitro in a single droplet in a microfluidic environment to mimic the environment in vivo for development of the embryo and to culture the embryos with good development and live births. Our results show that the dynamic culture powered with EWOD can manipulate a single droplet containing one mouse embryo and culture to the blastocyst stage. The rate of embryo cleavage to a hatching blastocyst with a dynamic culture is significantly greater than that with a traditional static culture (pculture of mouse embryos in a dynamic environment. To test the reproductive outcome of the embryos collected from an EWOD chip as a culture system, we transferred embryos to pseudo-pregnant female mice and produced live births. These results demonstrate that an EWOD-based microfluidic device is capable of culturing mammalian embryos in a microfluidic biological manner, presaging future clinical application. PMID:25933003

  3. Robert Edwards: the path to IVF

    OpenAIRE

    Johnson, Martin H

    2011-01-01

    The early influences on Robert Edwards’ approach to the scientific research that led to human IVF are described. His interest as a graduate student in the genetics of early mammalian development stimulated him later to investigate whether the origins of human genetic diseases such as Down, Klinefelter and Turner syndromes might be explained by events during egg maturation. This clinical problem provided the most powerful stimulus to achieve both oocyte maturation and fertilization in vitro in...

  4. Effect of paternal overweight or obesity on IVF treatment outcomes and the possible mechanisms involved

    Science.gov (United States)

    Yang, Qingling; Zhao, Feifei; Hu, Linli; Bai, Rui; Zhang, Nan; Yao, Guidong; Sun, Yingpu

    2016-01-01

    Leukocyte telomere lengths (LTLs) are shorter in obese compared with normal weight people. However, it is not known whether sperm telomere length (STL) is related to obesity. The aim of the study was to evaluate the impact of men’s body mass index (BMI) on STL, embryo quality, and clinical outcomes in couples undergoing IVF. In total, 651 couples were recruited, including 345 men with a normal BMI and 306 men with an overweight BMI (normal BMI group: 20–25 kg/m2; overweight BMI group: >28 kg/m2). We found that couples with male’s BMI over 28 kg/m2 exhibited a significantly lower fertilization rate, good-quality embryo rate and clinical pregnancy rate compared to their normal BMI counterparts. The mean STL in the overweight BMI group was also significantly shorter than that of the normal BMI group. The results also showed that individuals with higher BMI had higher ROS (Reactive oxygen species) content and sperm DNA fragmentation rate when compared with normal BMI individuals. Mitochondrial activity was also lower in the overweight BMI group than in the normal BMI group. This is the first report to find that STL is shorter in overweight/obese men, which may account for their poorer treatment outcomes in IVF cycles. PMID:27412918

  5. Aromatase inhibitors in stimulated IVF cycles

    Directory of Open Access Journals (Sweden)

    Tournaye Herman

    2011-06-01

    Full Text Available Abstract Aromatase inhibitors have been introduced as a new treatment modality that could challenge clomiphene citrate as an ovulation induction regiment in patients with PCOS. Although several randomized trials have been conducted regarding their use as ovulation induction agents, only few trials are available regarding their efficacy in IVF stimulated cycles. Current available evidence support that letrozole may have a promising role in stimulated IVF cycles, either when administered during the follicular phase for ovarian stimulation. Especially for women with poor ovarian response, letrozole appears to have the potential to increase clinical pregnancy rates when combined with gonadotropins, whereas at the same time reduces the total gonadotropin dose required for ovarian stimulation. However, given that in all of the trials letrozole has been administered in GnRH antagonist cycles, it is intriguing to test in the future how it may perform when used in GnRH agonist cycles. Finally administration of letrozole during luteal phase in IVF cycles offers another treatment modality for patients at high risk for OHSS taking into account that it drastically reduces estradiol levels

  6. Rol de la mitocondria y el estrés oxidativo en el bloqueo del desarrollo de embriones bovinos producidos in vitro Mitochondrial rol and oxidative stress in the developmental blockade of in vitro produced bovine embryos

    Directory of Open Access Journals (Sweden)

    AM Tarazona

    2010-01-01

    mediator of physiological and pathological states. Over the past years it has been shown that hydrogen peroxide (H2O2 is a pivoting molecule able to trigger cell death by different mechanisms that may or may not involve the transcription factors such as NFκB-p53, and is executed by effector caspases. It is believed that mitochondria may play an important role as a producer or as a target of H2O2, and as a mediator in apoptotic death of embryos. The purpose of this review is to present the state of the art about apoptosis triggered by oxidative stress and mediated by mitochondria in in vitro produced bovine embryos, as part of the explanation for the low efficiency in this process.

  7. The Birth and Routinization of IVF in China

    DEFF Research Database (Denmark)

    Wahlberg, Ayo

    2016-01-01

    How can it be that China today is home to some of the world’s largest IVF clinics, carrying out as many as 30,000 cycles annually? In this article, I address how IVF was developed in China during the early 1980s only to be routinized during the exact same period that one of the world’s most...... comprehensive family planning programmes aimed at preventing birth was being rolled out. IVF was not merely imported into China, rather it was experimentally developed within China into a form suitable for its restrictive family planning regulations. As a result, IVF and other assisted reproductive technologies...

  8. Avoiding multiple pregnancies in ART: a plea for single embryo transfer.

    Science.gov (United States)

    Gerris, J; Van Royen, E

    2000-09-01

    It has been generally accepted that triplets after IVF/intracytoplasmic sperm injection (ICSI) can and should be avoided by adopting a standard strategy of replacing no more than two embryos. However, there is an increasing awareness of the risks and costs and of the epidemic size of twin pregnancies after IVF/ICSI. This has resulted in efforts to replace no more than one embryo. However, this approach has been hampered by our relative inability to identify embryos with a very high implantation potential. To identify such embryos, a number of strategies are being considered, both at the two pronuclear (2PN), early cleavage and the blastocyst stages. At the 2PN stage, the polarity characteristics of the nucleoli have been shown to be correlated with a high implantation rate. Similarly, the morphological characteristics at day 2 and 3 have been used to describe top quality embryos in approximately 75% of all IVF/ICSI cycles. Blastocyst culture has resulted in very high implantation rates in the hands of some authors. No approach has shown its superiority at present, but initial experience with single embryo transfer (SET) at the early cleavage stage by Scandinavian and Belgian groups shows that an ongoing pregnancy rate of 35% and more can be achieved. Proper identification of patients at risk of a twin pregnancy after double embryo transfer is equally important. It is clear that mainly young patients (aged 80% of all twins) and are the main target group for twin prevention by SET of a top quality embryo at whatever stage. Therefore, in our opinion, although a further fine-tuning of both embryo and patient characteristics relating to a high risk for (twin) pregnancy is desirable, SET should be introduced carefully and progressively in each IVF/ICSI programme from now on. Correct counselling is very important and both public and private insurers will have to join in the discussion.

  9. 精子DNA碎片对IVF-ET结局的影响%Effects of sperm DNA fragmentation on outcomes of IVF-ET

    Institute of Scientific and Technical Information of China (English)

    张帅; 孟啸寅; 卓胜楠; 崔志英; 崔险峰; 张云山

    2013-01-01

    目的:探讨精子DNA碎片化率(DFI)对体外受精结局的影响.方法:采用改良精子染色质扩散实验(SCD)对139例接受体外受精(IVF)的男方进行精子DFI检测,并根据精子DFI值将上述IVF患者分为:A组(DFI<17.6%)、B组(17.6%≤DFI≤30%)、C组(DFI>30%),根据单因素方差分析统计方法分析各组IVF受精率、卵裂率、优质胚胎率和妊娠结局的差异.结果:与A组比较,B组和C组正常形态精子率、精子前向活动力[(a+b)%]、卵裂率、优质胚胎率均降低(P> 0.05 vs B组,P< 0.05 vs C组);与B组比较,C组正常形态精子率、精子前向活动力[(a+b)%]、卵裂率、优质胚胎率均降低(P<0.05);与A组比较,B组和C组受精率均降低(P< 0.05 vs B组,P< 0.05 vs C组);与B组比较,C组受精率降低(P<0.05);A、B、C3组间生化妊娠率、临床妊娠率差异无统计学意义(P> 0.05).结论:精子DNA碎片率升高不仅会导致精子前向活动力下降,还可导致IVF受精率、卵裂率、优质胚胎率下降,但对临床妊娠结局无明显影响.%Objective: To investigate the effects of sperm DNA fragmentation index on outcomes of IVF-ET. Methods: Sperm DNA fragmentation index of 139 infertile couples participating in IVF program were analyzed by sperm chromatin dispersion (SCD) test. They were divided into three groups upon the size of sperm DNA fragmentation index: Group A (DFI 30%). IVF outcomes were analyzed by one-way ANOVA, including of fertilization rate, cleavage rate, high quality embryos rate and pregnancy outcomes. Results: Compared with group A, normal morphology sperm rate and sperm forward motility [(a+b) %] as well as cleavage rate and high quality embryos rate were reduced in group B and group C (P>0.05 vs group B, P0.05 ). Conclusion: The increase in sperm DFI can not only result in an decrease in the sperm forward motility or normal morphology rate, but also lead to a decline of IVF fertilization rate, cleavage

  10. Comparing thaw survival, implantation and live birth rates from cryopreserved zygotes, embryos and blastocysts

    Directory of Open Access Journals (Sweden)

    Mary Ellen Pavone

    2011-01-01

    Full Text Available Context : Most in vitro fertilization (IVF programs employ embryo cryopreservation to enhance pregnancies from a single ovarian stimulation. More embryos are created, some of which are not transferred to the uterus immediately, generating a need for improved cryopreservation protocols. One protocol may involve growing embryos to a further stage of development, allowing only embryos with proven developmental capabilities to be cryopreserved. Here we examined thaw survival, implantation and live birth rates of embryos cryopreserved at different stages. Aims : We examined thaw survival, implantation and live birth rates of embryos cryopreserved at the zygote, day 3 (D3 embryos or blastocyst stage. Settings and Design : This is a retrospective study from a single academic IVF program. Patients and Methods :0 A retrospective study of all patients who had frozen embryos transferred to their uteri from year 2002 to 2008 at a single academic IVF program was conducted. Statistical Analysis Used : Analysis of variance followed by Fisher′s Exact Test was performed to compare the survival after thaw, implantation and live birth rates between the three groups. Results : One thousand nine hundred and ninety-one zygotes, 2880 D3 embryos and 503 blastocysts were frozen using a slow freeze technique, thawed and transferred. Significantly more D3 embryos and blastocysts survived the thawing process compared to zygotes and significantly higher implantation rate per number of thawed blastocysts was achieved than that for zygotes. Live birth rates were similar between the three groups. Conclusions : Growing embryos to blastocyst stage prior to cryopreservation is associated with fewer frozen embryos but does not appear compromise patients′ chance of achieving pregnancy

  11. Assessment of Uterine Receptivity by Endometrial and Subendometrial Blood Flows Measured by Vaginal Color Doppler Ultrasound in Women Undergoing IVF Treatment

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Obiective To evaluate endometrial and Subendometrial blood flows measured by vaginal color Doppler ultrasound as a predicator of endometrial receptivity in women undergoing IVF treatment.Methods A total of 119 infertile patients undergoing the first IVF/ICSI-ET cycle were recruited.Three groups were divided according to a color Doppler ultrasound examination performed on the day of hCG injection.Group A,endometrial and subendometrial blood flows were 2 branches and below,group B,endometrial and subendometrial blood flows were between 3 and 4 branches;group C, endometrial and subendometrial blood flows were 5 branches and above.Patients were transferred 1-3 embryos each.Demographic data,ovarian responses.endometrial thickness,PI,RI,development of embryo and IVF result among groups were compared.Results Demographic data,ovarian responses,endometrial thickness,PI,RI and development of embryo among groups have no significant difference.The pregnancy rate of group A was significantly lower than that of group B(P<0.05) and group C(P<0.01).The implantation rate of group A was significantly lower than than of group C (P<0.01).There was no significant difference of the rate of pregnancy and implantation between group B and group C (P>0.05).Conclusion Endometrial and subendometrial blood flows measured by vaginal color Doppler ultrasound is a good predicator of pregnancy during IVF treatment.A good endometrial and subendometrial blood flows is benefit for the result of IVF.

  12. Single embryo transfer - state of the art.

    Science.gov (United States)

    De Neubourg, Diane; Gerris, Jan

    2003-12-01

    Every practitioner active in the field of assisted reproduction treatment is aware of the risks and complications related to twin and higher-order multiple pregnancies. Introduction of single embryo transfer (SET) into IVF/intracytoplasmic sperm injection (ICSI) is one of the possible ways of reducing the rate of twin pregnancy. Careful selection of patients, in combination with elective SET, has been shown to decrease the twin pregnancy rate while maintaining a stable ongoing pregnancy rate. The combination of a woman younger than 38 years of age, in her first or second IVF/ICSI cycle and with an embryo with a high implantation potential is the key to successful SET. This article will discuss embryo selection and patient selection and review the data published on SET. In the Centre for Reproductive Medicine at Middelheim Hospital, 39% of all transfers in 2002 were SET; the ongoing pregnancy rate remained stable at 30.6%. The twin (multiple) pregnancy rate declined to 11.7%. Particular attention should be drawn to the augmenting effect of the pregnancy rate of frozen-thawed cycles. Health economic data available so far subscribe the plea for SET.

  13. Effect of insulin-like growth factor-1 during in vitro oocyte maturation and in vitro culture of bovine embryos Efeito do fator de crescimento semelhante à insulina-1 durante a maturação in vitro dos oócitos e cultivo in vitro de embriões bovinos

    OpenAIRE

    M.D. Quetglas; L.A Coelho; Garcia, J M; E.B. Oliveira Filho; C.R. Esper

    2001-01-01

    The effects of insulin-like growth factor-I (IGF-I) on in vitro maturation (IVM) (experiment I) and on in vitro embryo development (experiment II) of bovine oocytes fertilized in vitro, were evaluated in terms of cleavage (CR), blastocyst (BR) and hatching (HR) rates. For IVM, immature cumulus-oocyte complexes were cultured in TCM-199 medium supplemented with Hepes, sodium bicarbonate, sodium pyruvate, additives, fetal calf serum (B-199 medium) and gonadotropins (14 U/ml PMSG and 7 U/ml hCG)....

  14. Blastocyst development potential of D3 low quality embryos

    Institute of Scientific and Technical Information of China (English)

    Zhen Jing-ran; Sun Zhen-yi; Yu Qi; Deng Cheng-yan; Zhou Yuan-zheng; He Fang-fang

    2011-01-01

    Objective:To investigate the potential of blastocyst development of D3 low quality embryos for blastocyst culturing and freezing.Methods:Two thousand one hundred and eighty embryos of 398 IVF/ICSI patients' were observed after D3 embryo transfer.The low-quality embryos were cultured by sequential micro-drops and continuously observed blastocyst formation from D3 to D6.Results:(1) A total of 1,546 low-quality embryos were collected,and 426 blastocysts formed in DS-D6.Total blastocyst formation rate (BFR) was 27.56%,of which embryos with grade 6 Ⅲ/Ⅳ were 40.87 % (318/778),5 Ⅰ /Ⅱ 28.85% (30/104),4Ⅰ/Ⅱ 8.88% (16/180),4Ⅲ/IV 19.72% (56/284),2-3 Ⅰ/Ⅱ 3% (6/200).(2) Embryos with more blastomeres had a higher blastocyst formation rate.The lower qualities of embryos resulted in lower blastocyst formation rate.Conclusions:Low-quality embryos still have the potential to develop into blastocysts.Therefore,they should not be discarded on D3.They should be continually cultured to D6 in order to reduce embryo wastage and get good clinical results.

  15. The associated factors to endometrial cavity fluid and the relevant impact on the IVF-ET outcome

    Directory of Open Access Journals (Sweden)

    Li Ya-Qiong

    2010-05-01

    Full Text Available Abstract Background Endometrial cavity fluid (ECF is a fluid accumulation within the endometrial cavity. The significance of ECF remains unclear during the program of in vitro fertilization-embryo transfer (IVF-ET. The aim of the present study was to investigate the associated factors to ECF, visualized through ultrasound at the day of oocyte retrieval, and the relevant impact on the outcome of IVF-ET. Methods From the clinical data of 1557 infertility patients for IVF-ET program, 46 ECF patients were retrospectively selected as the ECF group; and another 134 patients with a bilateral salpingectomy and without ECF, selected as the control group. The demographics and the outcome of IVF-ET were compared between the two groups. Results The incidence of ECF was 2.95% (46/1557. Over half (28/46, 60.87% of ECF patients had tubal infertility. Only 12 Of 46 ECF patients (26.09% had visible hydrosalpinx on ultrasonography before ovarian stimulation. The cycle cancellation rate (4/46, 8.69% of ECF group was not significantly higher than that of the control group (6/134, 4.48%; P > 0.05. Reasons for cycle cancellation in both groups were all the high risk of ovarian hyperstimulation syndrome (OHSS. No significant difference was found in clinical pregnancy rate between the patients with their ECF 0.05. No clinical pregnancy was found among those patients with their ECF equal or higher 3.5 mm in APD. Conclusions It was tubal infertility, not hydrosalpinx, which was related to the development of ECF. Excessive ECF (equal or higher 3.5 mm in APD at the day of oocyte retrieval would have a negative impact on the outcome of IVF-ET.

  16. Association between serum estradiol level on the hCG administration day and IVF-ICSI outcome

    Directory of Open Access Journals (Sweden)

    Tayfun Kutlu

    2012-01-01

    Full Text Available Background: Estradiol (E2 is required for follicular development and play an important role in embryo implantation. Objective: The aim of this study was to assess the impact of serum E2 levels on the day of hCG administration in IVF-ICSI patients who are performed controlled ovarian hyperstimulation (COH.Materials and Methods: A total of 203 women who were undergone one time IVF cyclus were evaluated in this cross sectional study. All the patients were treated either with long protocol or with microdose flare protocol. The patients were categorized into five groups according to the serum E2 levels on the day of hCG administration.Results: The mean number of the retrieved oocytes was (NRO 10.6±6.7, mean fertilization rate was 55.7±24.8, and implantation rate was 9.0±19.2. Of 203 patients, 43 (21% patients were pregnant. When the overall results are examined, the number of the retrieved oocytes and the number of transferred embryos were better in patients with serum E2 levels >4000 pg/ml and these values were statistically significant. There were no statistical difference in patients 37 years or older. In women ≤36 years old, the IVF-ICSI outcomes were better in patients with serum E2 levels >4000 pg/ml.Conclusion: In spite of the lack of high quality evidence to support a positive association between serum E2 levels and IVF-ICSI outcomes, this study shows that high E2 levels during COH might be associated with an increased potential of pregnancy depending on better ovarian response. When the overall results are examined, the best scores were in patients with serum E2 levels >4000 pg/ml.

  17. Perinatal outcome in singletons after modified natural cycle IVF and standard IVF with ovarian stimulation

    NARCIS (Netherlands)

    M.J. Pelinck; M.H. Keizer; A. Hoek; A.H.M. Simons; K. Schelling; K. Middelburg; M.J. Heineman

    2010-01-01

    Objective: Singletons born after IVF treatment are at risk for adverse pregnancy outcome, the cause of which is unknown. The aim of the present study was to investigate the influence of ovarian stimulation on perinatal outcome. Study design: In this single-centre retrospective study, perinatal outco

  18. Use of sexed sorted semen for fixed-time artificial insemination or fixed-time embryo transfer of in vitro-produced embryos in cattle.

    Science.gov (United States)

    Pellegrino, C A G; Morotti, F; Untura, R M; Pontes, J H F; Pellegrino, M F O; Campolina, J P; Seneda, M M; Barbosa, F A; Henry, M

    2016-08-01

    Artificial insemination and in vitro embryo production are powerful tools for disseminating superior genetic qualities and improving the reproductive performance of dairy and beef cattle. In conjunction with these biotechnologies, sexed-sorted semen has been used to obtain offspring of a predetermined sex. This study compared the pregnancy rates obtained using in vitro fertilization/timed embryo transfer (IVF/TET) and timed artificial insemination (TAI), both performed using sexed-sorted (Y-chromosome-bearing) semen obtained from the same bull. For the in vitro embryo production, the ovaries of 250 Nelore cows with known histories were collected in the slaughterhouse and used for IVF. After evaluation of the recipients (IVF/TET group; n = 974), the resultant embryos were transferred to the females with corpus luteum (n = 822). The pregnancy-related data for this group were compared with those for the TAI group (n = 974). Ultrasonography was performed at 60 days to determine the pregnancy status and confirm the sex of the fetus. A total of 2008 oocytes produced 1050 embryos, with 52% of them reaching the blastocyst stage. The pregnancy rate and the accuracy in determining the fetal sex were 35.4% (345/974) and 95.07% (328/345), respectively, for the IVF/TET group and 30% (293/974; P < 0.05) and 94.88% (278/293), respectively, for the TAI group. In the present study, we concluded that male calves could be better obtained using IVF/TET rather than TAI; therefore, this strategy can be considered to increase the pregnancy rate of beef cattle. PMID:27068357

  19. Impact of GnRH analogues on oocyte/embryo quality and embryo development in in vitro fertilization/intracytoplasmic sperm injection cycles: a case control study

    Directory of Open Access Journals (Sweden)

    Rigó János

    2009-09-01

    Full Text Available Abstract Background Despite the clinical outcomes of ovarian stimulation with either GnRH-agonist or GnRH-antagonist analogues for in vitro fertilization (IVF being well analysed, the effect of analogues on oocyte/embryo quality and embryo development is still not known in detail. The aim of this case-control study was to compare the efficacy of a multiple-dose GnRH antagonist protocol with that of the GnRH agonist long protocol with a view to oocyte and embryo quality, embryo development and IVF treatment outcome. Methods Between October 2001 and December 2008, 100 patients were stimulated with human menopausal gonadotrophin (HMG and GnRH antagonist in their first treatment cycle for IVF or intracytoplasmic sperm injection (ICSI. One hundred combined GnRH agonist + HMG (long protocol cycles were matched to the GnRH antagonist + HMG cycles by age, BMI, baseline FSH levels and by cause of infertility. We determined the number and quality of retrieved oocytes, the rate of early-cleavage embryos, the morphology and development of embryos, as well as clinical pregnancy rates. Statistical analysis was performed using Wilcoxon's matched pairs rank sum test and McNemar's chi-square test. P Results The rate of cytoplasmic abnormalities in retrieved oocytes was significantly higher with the use of GnRH antagonist than in GnRH agonist cycles (62.1% vs. 49.9%; P Conclusion Antagonist seemed to influence favourably some parameters of early embryo development dynamics, while other morphological parameters seemed not to be altered according to GnRH analogue used for ovarian stimulation in IVF cycles.

  20. Non-invasive assessment of in-vitro embryo quality to improve transfer success

    OpenAIRE

    Højbøge, Tina Rødgaard; Peter M H Heegaard; Callesen, Henrik

    2015-01-01

    Although IVF has been performed routinely for many years to help couples with fertility problems and in relation to modern breeding of farm animals, pregnancy rates after transfer to a recipient have not improved during the last decade. Early prediction of the viability of in-vitro developed embryos before the transfer to a recipient still remains challenging. Presently, the predominant non-invasive technique for selecting viable embryos is based on morphology, where parameters such as rates ...

  1. Consequences of vanishing twins in IVF/ICSI pregnancies

    DEFF Research Database (Denmark)

    Pinborg, Anja Bisgaard; Lidegaard, Ojvind; la Cour Freiesleben, Nina;

    2005-01-01

    Spontaneous reductions are a possible cause of the increased morbidity in IVF singletons. The aim of this study was to assess incidence rates of spontaneous reductions in IVF/ICSI twin pregnancies and to compare short- and long-term morbidity in survivors of a vanishing co-twin with singletons...

  2. Optimizing the culture environment and embryo manipulation to help maintain embryo developmental potential.

    Science.gov (United States)

    Swain, Jason E; Carrell, Doug; Cobo, Ana; Meseguer, Marcos; Rubio, Carmen; Smith, Gary D

    2016-03-01

    With increased use of comprehensive chromosome screening (CCS), the question remains as to why some practices do not experience the same high levels of clinical success after implementation of the approach. Indeed, the debate surrounding the efficacy and usefulness of blastocyst biopsy and CCS continues. Importantly, several variables impact the success of an assisted reproductive technology cycle. Transfer of a euploid embryo is but one factor in an intricate system that requires numerous steps to occur successfully. Certainly, the culture environment and the manipulations of the embryo during its time in the laboratory can impact its reproductive potential. Environmental stressors ranging from culture media to culture conditions and even culture platform can impact biochemical, metabolic, and epigenetic patterns that can affect the developing cell independent of chromosome number. Furthermore, accompanying procedures, such as biopsy and vitrification, are complex and, when performed improperly, can negatively impact embryo quality. These are areas that likely still carry room for improvement within the IVF laboratory.

  3. Anti-Mullerian hormone : a marker for oocyte quantity, oocyte quality and embryo quality?

    NARCIS (Netherlands)

    Fong, S. Lie; Baart, E. B.; Martini, E.; Schipper, I.; Visser, J. A.; Themmen, A. P. N.; de Jong, F. H.; Fauser, B. J. C. M.; Laven, J. S. E.

    2008-01-01

    Serum anti-Mullerian hormone (AMH) concentrations decline with increasing age and constitute a Sensitive marker for ovarian ageing. In addition, basal serum AMH concentrations predict ovarian response during IVF cycles. Concomitantly, oocyte quantity and embryo quality decrease with advancing age. H

  4. What about the remaining twins since single-embryo transfer? How far can (should) we go?

    Science.gov (United States)

    De Neubourg, D; Gerris, J

    2006-04-01

    Single-embryo transfer (SET) and more specifically elective SET (eSET) have taken their place in good clinical IVF/ICSI practice. After the initial cautious search for the characteristics of the twin-prone patient and of the selection of the embryo with the highest implantation potential many centres have embarked on the (progressive) implementation of SET, either by conviction or forced by legislation or both. It was only because the ongoing pregnancy rates remained largely unaffected that SET was accepted. Generally speaking, it can be said that the twinning rate after IVF/ICSI has dropped by at least 50% simply by transferring only one good-quality embryo in the first and second fresh IVF/ICSI cycles in young women, without decrease in the overall pregnancy rate. Preventing 'the second half' of IVF/ICSI twins constitutes another and probably tougher challenge because the target group is a heterogeneous mix consisting of patients in very different clinical situations. Can we expand our experience for further twin prevention to women of older age and to cycles of higher rank without a significant drop in pregnancy rates? Can we extend it to more cryopreservation cycles? To have an idea of future target groups for increased application of SET, we analysed the remaining twins after double-embryo transfer (DET), and from these data we suggest expanding the eSET policy to women <38 years of age until the third cycle and to cryopreservation cycles.

  5. First successful pregnancies following embryo selection using Time-lapse technology in Iran: Case report

    Directory of Open Access Journals (Sweden)

    Azita Faramarzi

    2015-03-01

    Full Text Available Background: Embryo selection is a vital part of in vitro fertilization (IVF programs, with morphology-based grading systems having been widely used for decades. Time-lapse imaging combined with embryo morph kinetics may proffer a non-invasive means for improving embryo selection. We report the first ongoing and chemical pregnancies using Time-lapse embryo scope to select best embryos for transfer in Iran. Cases: A case with tubal factor infertility was admitted to IVF program with normozoospermia. After ovarian hyper stimulation, 6 COCs were retrieved and inseminated with 25,000 progressive sperms/ oocyte. Five zygotes were placed individually into the micro wells of equilibrated embryo scope dish for Time-lapse observation, and incubated at 37°C, 5% CO2. On day 3, single embryo transfer (SET took place based on kinetic parameters of the embryos. Clinical pregnancy was confirmed 7 weeks after SET. The second case with history of previous ICSI failure was admitted with azoospermia. Nine MII oocytes underwent ICSI, and incubated in Time-lapse facilities. The rest of procedures were followed as described for case 1. Chemical pregnancy was confirmed 15 days after SET. Conclusion: This approach opens a way to select best embryo non-invasively for SET; thus, increasing implantation, while reducing multiple pregnancy complications.

  6. Comparison of ultrasound-guided endometrial polypectomy carried out on the oocyte retrieval day and the first day of ovarian stimulation in IVF-ICSI cycles.

    Science.gov (United States)

    Moon, Jei-Won; Kim, Chung-Hoon; Park, So-Yun; Kim, Sung-Hoon; Chae, Hee-Dong; Kang, Byung-Moon

    2016-09-01

    In this retrospective cohort study, the effect of endometrial polypectomy carried out on the day of oocyte retrieval and on the first day of ovarian stimulation in patients with a large (≥10 mm) endometrial polyp undergoing IVF and intractyoplasmic sperm injection (ICSI) was investigated and compared. A total of 74 eligible IVF-ICSI cycles in 74 women who underwent endometrial polypectomy on either the day of oocyte pick-up (late polypectomy group, 39 cycles) or the first day of ovarian stimulation (early polypectomy group, 35 cycles) between January 2007 and July 2012 were included in this study. Patient characteristics between early and late polypectomy groups were similar. Total dose and days of recombinant human FSH administered, numbers of retrieved oocytes, mature oocytes, fertilized oocytes, grade 1 or 2 embryos and embryos transferred between the two groups were also similar, as was clinical pregnancy rate per cycle, embryo implantation rate and spontaneous abortion rate between the two groups. Therefore, endometrial polypectomy on the day of oocyte retrieval could be a more patient-friendly option for patients with a large endometrial polyp undergoing IVF-ICSI.

  7. Comparison of the Effects of Nest PCR System Established by Sry Gene and Y Chromosome Repeated Sequence for Sex Determination of Bovine Embryo%Sry基因和Y染色体重复序列在牛早期胚胎性别鉴定中应用效果的比较

    Institute of Scientific and Technical Information of China (English)

    张伟; 王世银; 张兆旺; 赵兴绪

    2011-01-01

    Sty gene and Y chromosome repeated sequence were selected as male-specific gene for sex determination of bovine embryo in this study. Eight pairs of primers were designed and a nest PCR system was established in order to compare the amplification effect. The results showed that the system established by Y chromosome repeated sequence was more sensitive and stable than the one established by Sty gene when the PCR template was the DNA from one embryonic ceil, so it was more suitable for sex determination of bovine embryo.%本试验以牛Sty基因和Y染色体重复序列作为雄性特异性基因,分别设计引物,建立多重巢式PCR体系,比较二者在牛早期胚胎性别鉴定中的应用效果。试验结果表明,当扩增体系中的模板量为一个胚胎细胞的DNA量时,以Y染色体重复序列构建的扩增体系比Sry基因具有更高的灵敏度和稳定性,更适合用于牛早期胚胎性别鉴定。

  8. Can Comprehensive Chromosome Screening Technology Improve IVF/ICSI Outcomes? A Meta-Analysis.

    Directory of Open Access Journals (Sweden)

    Minghao Chen

    Full Text Available To examine whether comprehensive chromosome screening (CCS for preimplantation genetic screening (PGS has an effect on improving in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI outcomes compared to traditional morphological methods.A literature search was conducted in PubMed, EMBASE, CNKI and ClinicalTrials.gov up to May 2015. Two reviewers independently evaluated titles and abstracts, extracted data and assessed quality. We included studies that compared the IVF/ICSI outcomes of CCS-based embryo selection with those of the traditional morphological method. Relative risk (RR values with corresponding 95% confidence intervals (CIs were calculated in RevMan 5.3, and subgroup analysis and Begg's test were used to assess heterogeneity and potential publication bias, respectively.Four RCTs and seven cohort studies were included. A meta-analysis of the outcomes showed that compared to morphological criteria, euploid embryos identified by CCS were more likely to be successfully implanted (RCT RR 1.32, 95% CI 1.18-1.47; cohort study RR 1.74, 95% CI 1.35-2.24. CCS-based PGS was also related to an increased clinical pregnancy rate (RCT RR 1.26, 95% CI 0.83-1.93; cohort study RR 1.48, 95% CI 1.20-1.83, an increased ongoing pregnancy rate (RCT RR 1.31, 95% CI 0.64-2.66; cohort study RR 1.61, 95% CI 1.30-2.00, and an increased live birth rate (RCT RR 1.26, 95% CI 1.05-1.50; cohort study RR 1.35, 95% CI 0.85-2.13 as well as a decreased miscarriage rate (RCT RR 0.53, 95% CI 0.24-1.15; cohort study RR 0.31, 95% CI 0.21-0.46 and a decreased multiple pregnancy rate (RCT RR 0.02, 95% CI 0.00-0.26; cohort study RR 0.19, 95% CI 0.07-0.51. The results of the subgroup analysis also showed a significantly increased implantation rate in the CCS group.The effectiveness of CCS-based PGS is comparable to that of traditional morphological methods, with better outcomes for women receiving IVF/ICSI technology. The transfer of both trophectoderm-biopsied and

  9. Nucleolar remodeling in nuclear transfer embryos

    DEFF Research Database (Denmark)

    Laurincik, Jozef; Maddox-Hyttel, Poul

    2007-01-01

    to develop fully functional nucleoli. In bovine in vivo developed embryos, a range of important nucleolar proteins (e.g., topoisomerase I, upstream binding factor and RNA polymerase I, fibrillarin, nucleophosmin and nucleolin) become localized to the nucleolar anlage over several cell cycles. This relocation...

  10. Maternal death related to IVF in the Netherlands 1984-2008.

    NARCIS (Netherlands)

    Braat, D.D.M.; Schutte, J.M.; Bernardus, R.E.; Mooij, T.M.; Leeuwen, F.E. van

    2010-01-01

    BACKGROUND: We assessed all deaths in the Netherlands that might have been related to IVF or to an IVF pregnancy in order to investigate this most serious complication. METHODS: All deaths related to IVF, within 1 year after IVF, from 1984 to 2008 were collected by sending a letter to all gynaecolog

  11. Effects of Bushen Tiaozhou on IVF-ET Intervention and Results%补肾调周法对IVF-ET的干预及其结局的影响

    Institute of Scientific and Technical Information of China (English)

    刘建国; 金保方; 杨晓玉; 张新东

    2012-01-01

    OBJECTIVE To study Bushen Tiaozhou method and its intervetion in the feasibility and effectiveness of IVF-ET. To explore its value of improving IVF-ET success rate. METHODS 160 IVF-ET patients were randomly divided into observation group (80 cases) and control group (80 cases). Patients in control group were given regular treatment from Clinical Reproductive Medical Center of Jiangsu Province People's Hospital and patients in observation groups were given extra Bushen Tiaozhou Method, The endometrial thickness, number of oocytes, fertilization rate, good embryo rate, biochemical pregnancy rate and clinical pregnancy rate were observed two weeks later. RESULTS After Bushen Tiaozhou method, the endometrial thickness, number of oocytes, fertilization rate, good embryo rate, biochemical pregnancy rate and clinical pregnancy rate were significantly changed, especially the endometrial thickness (P<0.01) and fertilization rate (P<0.01). It also significantly improved number of oocytes, good embryo rate, final clinical pregnancy rate, compared to the control group (P<0.05). CONCLUSION Bushen Tiaozhou method can promote ovulation number, increase endometrial thickness, improve fertilization rates and quality embryo rate, assist embryo implantation tocolysis and improve the clinical pregnancy rate.%目的 研究补肾调周法干预体外受精-胚胎移植技术(IVF-ET)可行性和有效性.探讨补肾调周法提高IVF-ET成功率的价值及意义.方法 将160例接受IVF-ET的患者随机分为观察组(80例)和对照组(80例),对照组采用江苏省人民医院生殖中心常规方案,观察组在对照组治疗的基础上运用补肾调周法进行中药干预,观察2组患者子宫内膜厚度、获卵数、受精率、优质胚胎率、生化妊娠率和临床妊娠率.结果 经补肾调周法干预后,观察组可以极显著提高患者子宫内膜的厚度(P<0.01)及受精率(P<0.01);而且在提高获卵数、优质胚胎率及最终提高临床妊娠

  12. Cell viability of bovine spermatozoa subjected to DNA electroporation and DNAse I treatment.

    Science.gov (United States)

    Cavalcanti, Paulo Varoni; Milazzotto, Marcella Pecora; Simões, Renata; Nichi, Marcilio; de Oliveira Barros, Flavia Regina; Visintin, Jose Antonio; Assumpção, Mayra Elena Ortiz D'Avila

    2016-04-15

    Many mechanisms involved in sperm-mediated gene transfer (SMGT) are still unknown. It is still a matter of debate whether exogenous DNA fragments incorporated by the embryo are originated from those bound to the sperm membrane or by those that penetrated the intracellular compartment. In an attempt to elucidate the transmission mechanism of exogenous DNA molecules by sperm, some authors suggested a treatment with DNAse I to remove DNA molecules outside the sperm. But little is known regarding the effects of DNAse I treatment on sperm viability and its impact on sperm organelles. An important aspect of the SMGT technique is the amount of exogenous DNA incubated with sperm, which may influence the internalization rate. Due to the inconsistencies found in literature, this work aimed to contribute to bovine sperm physiology knowledge evaluating the effects of different DNA concentrations, electroporation, and DNAse I treatments on sperm viability characteristics, DNA uptake, and IVF. For that, the effects of different concentrations of exogenous DNA (250, 500 and 1000 ng/10(6) cells) and incubation or electroporation were tested on sperm functional characteristics and in vitro embryo production. No effect of DNA concentration was observed on uptake, plasma membrane integrity, and mitochondrial membrane potential. The addition of exogenous DNA induced a decrease on acrosomal lesion in the 500-ng group when compared to the control. Cells incubated with DNA, electroporated, and treated with DNAse I presented a deleterious influence on mitochondrial membrane potential. In vitro fertilization was made with 1000 ng of DNA, sperm cells incubated or electroporated followed by DNAse I treatment. No significant difference was found in cleavage rate. Blastocyst rates were 24.36% for the control; 19.65% for incubated; 3.5% for electroporated control; and 17.40% for electroporated. There is a significant difference in blastocyst rate between the control and electroporated

  13. The mental state of women with an IVF pregnancy.

    Directory of Open Access Journals (Sweden)

    Zakharova E.I.

    2015-03-01

    Full Text Available An in vitro fertilization (IVF pregnancy is stressful both financially and emotionally. Patients undergoing an IVF procedure often have already had infertility and reproductive losses. Pregnancy through IVF involves the increased risk of various medical complications. Experts around the world are actively engaged in studying the specifics of the mental state of participants in IVF programs during pregnancy. Of critical importance is the issue of providing psychological support for couples who are preparing for and who have received an IVF pregnancy. The aim of our research was to investigate the mental state of women participating in an IVF program. The study involved 224 pregnant women in the second and third trimesters: 62 women with an IVF pregnancy and 162 women who conceived naturally. The study took place at the Kulakov Scientific Center for Obstetrics, Gynecology, and Perinatology in Moscow, Russia. All the study participants had encountered medical complications during their pregnancy. No significant differences were identified in mental well-being in the two groups; this finding suggests that somatic complications during pregnancy are a general source of anxiety regardless of the reason for their occurrence. The second and third trimesters of pregnancy register increased anxiety levels associated with experiences of reproductive loss and the presence of physical problems. The main resources of a woman’s personality that contribute to her self-confidence and mental stability are her professional employment and flexible behavior.

  14. In vitro fertilization in Japan — Early days of in vitro fertilization and embryo transfer and future prospects for assisted reproductive technology —

    Science.gov (United States)

    SUZUKI, Masakuni

    2014-01-01

    Assisted reproductive technology (ART) such as in vitro fertilization (IVF) and embryo transfer (ET) has been essential in the treatment of infertility. The world’s first IVF-ET baby was born in 1978 based on the technique developed by Dr. Robert Edwards and Dr. Patrick Steptoe.1) In Japan, the first IVF-ET birth was reported in 1983 by Prof. Masakuni Suzuki at Tohoku University School of Medicine.2,3) IVF-ET is a procedure used to achieve pregnancy that consists of extracting oocytes from an infertile woman, fertilizing them in vitro, and transferring fertilized eggs into the patient’s uterine cavity (Fig. 1). Since the first report of successful IVF-ET, numerous techniques related to ART, such as cryopreservation of oocytes and embryos, gamete intrafallopian transfer (GIFT), and microinsemination, have been developed and refined (Table 1). Herein we describe the history of basic research in IVF-ET that led to human applications, how the birth of the first IVF-ET baby was achieved in Japan, the current status of ART in Japan, issues related to ART, and future prospects for ART. PMID:24814992

  15. In vitro fertilization in Japan - early days of in vitro fertilization and embryo transfer and future prospects for assisted reproductive technology.

    Science.gov (United States)

    Suzuki, Masakuni

    2014-01-01

    Assisted reproductive technology (ART) such as in vitro fertilization (IVF) and embryo transfer (ET) has been essential in the treatment of infertility. The world's first IVF-ET baby was born in 1978 based on the technique developed by Dr. Robert Edwards and Dr. Patrick Steptoe. In Japan, the first IVF-ET birth was reported in 1983 by Prof. Masakuni Suzuki at Tohoku University School of Medicine. IVF-ET is a procedure used to achieve pregnancy that consists of extracting oocytes from an infertile woman, fertilizing them in vitro, and transferring fertilized eggs into the patient's uterine cavity (Fig. 1). Since the first report of successful IVF-ET, numerous techniques related to ART, such as cryopreservation of oocytes and embryos, gamete intrafallopian transfer (GIFT), and microinsemination, have been developed and refined (Table 1). Herein we describe the history of basic research in IVF-ET that led to human applications, how the birth of the first IVF-ET baby was achieved in Japan, the current status of ART in Japan, issues related to ART, and future prospects for ART. PMID:24814992

  16. 激光辅助孵化在前次IVF-ET失败患者中的应用%Application of LAH in patient with previous IVF-ET failure cycle

    Institute of Scientific and Technical Information of China (English)

    罗亚宁; 肖西峰; 王晓红; 李丽; 刘赛; 周婷

    2008-01-01

    [目的]探讨激光辅助孵化(1aser-assisted hatching,LAH)对前次体外受精.胚胎移植(ill vitro fertili-zation-embryo transfer,IVF-ET)失败患者的临床妊娠率和胚胎种植率的影响.[方法]对前次IVF-ET失败并排除子宫内膜、胚胎质量等因素的90例患者,根据是否采用激光辅助孵化分为LAH组和对照组,对LAH组患者的胚胎采用部分透明带薄化后进行移植.比较两组的临床妊娠率和胚胎种植率.[结果]LAH组和对照组的临床妊娠率分别为48.8%(20,41)和34.7%(17/49)(P>0.05);胚胎种植率分别为25.8%(30/116)和15.7%(22/140)(P<0.05);两组比较胚胎种植率有差异.[结论]激光辅助孵化可以提高前次IVF-ET失败患者的胚胎种植率.

  17. Embryo apoptosis identification: Oocyte grade or cleavage stage?

    Science.gov (United States)

    Bakri, Noraina Mohd; Ibrahim, Siti Fatimah; Osman, Nurul Atikah; Hasan, Nurhaslina; Jaffar, Farah Hanan Fathihah; Rahman, Zulaiha Abdul; Osman, Khairul

    2015-01-01

    Apoptosis is a programed cell death that is vital for tissue homeostasis. However, embryo apoptosis had been known to be related to embryo fragmentation which should be avoided in in vitro fertilization (IVF). The purpose of this study was to evaluate the relationship of embryo apoptosis with the grade of immature oocytes and cleavage stage of in vitro produced (IVP) cattle embryos. This study consisted of 345 oocytes collected through ovary slicing. Immature oocytes were graded as A, B and C. This grading was based on cumulus cell thickness and compactness. All oocytes then underwent an in vitro maturation (IVM) procedure. An IVF was done 24 h after IVM culture. Prior to staining, stage of cleaved embryos was determined and classified as either 2, 4, 8 or >8-cell embryo stage. Apoptosis status of cleaved IVP embryos was determined by using annexin V-FITC staining technique at 48 and 72 h post insemination (hpi). Apoptosis status for each embryo was classified as either early or late. The result showed that there was no significant difference (p > 0.05) of apoptosis status among grade A, B and C embryos. All grades of oocytes showed embryo apoptosis where 1.5% late apoptosis for grade A, 4.5% and 10.4% of early and late apoptosis for grade B and grade C. Early apoptosis was not seen in grade A embryo. We also noted no significant difference (p > 0.05) of apoptosis status between 2, 4, 8 and >8-cell embryo stage. Early apoptosis was also not seen in >8-cell stage. Even though there were no differences in apoptosis expression between the three classes, the cleavage rate of grade A oocytes was significantly higher (p < 0.01) than grade B and grade C. In conclusion, the apoptosis expression in the embryo can occur regardless of the oocyte quality and the cleavage stage of the embryo produced. PMID:26858565

  18. Efeito do citrato e taurina em meio CR2aa no desenvolvimento de embriões bovinos fecundados in vitro Effect of citrate and taurine added to CR2aa medium on the development of in vitro-fertilized bovine embryos

    Directory of Open Access Journals (Sweden)

    L.S.A. Camargo

    2009-02-01

    Full Text Available Avaliou-se o efeito do citrato em meio CR2aa suplementado com soro fetal bovino (SFB ou livre de proteínas séricas e sua associação com taurina no desenvolvimento de embriões bovinos fecundados in vitro. Embriões foram cultivados em CR2aa contendo 0, 0,5, 1,0 e 3,0mM citrato, suplementado com 10% SFB (experimento 1 ou com álcool polivinil (PVA; experimento 2. No terceiro experimento, embriões foram cultivados em meio com 0,5mM citrato, ou 7mM taurina, ou com a associação de ambos, suplementado com SFB. Os cultivos foram realizados com células do cumulus em ambiente a 38,8ºC com 5% de CO2 em ar atmosférico. Melhora no desenvolvimento embrionário foi observado no cultivo de embriões em CR2aa com 0,5 e 1,0mM citrato na ausência de SFB (P0,05 a produção de embriões ou o número de células. Citrato em meio CR2aa pode ser uma alternativa para cultivo embrionário em condições atmosféricas com 5% de CO2 em ar na ausência de proteína sérica.The effect of citrate added to CR2aa medium supplemented with fetal calf serum (FCS or serum-proteinfree and its association with taurine on the development of in vitro-fertilized bovine embryos was evaluated. Embryos were cultured with 0, 0.5, 1.0, and 3.0mM citrate, in CR2aa supplemented with 10% FCS (experiment 1, or polyvinyl alcohol (PVA; experiment 2. In experiment 3, embryos were cultured with 0.5mM citrate, 7.0mM taurine or with association of both, in medium supplemented with FCS. Embryo culture was performed with cumulus cells at 38.8ºC in 5% CO2 under air for all experiments. Positive effect on embryo development was only observed with 0.5 and 1.0mM citrate in FCS-free CR2aa (P0.05 embryo rate nor total cell number. Citrate in CR2aa medium can be an alternative for serumfree embryo culture under 5% CO2 in air, absence of serum protein.

  19. Resurgence of Minimal Stimulation In Vitro Fertilization with A Protocol Consisting of Gonadotropin Releasing Hormone-Agonist Trigger and Vitrified-Thawed Embryo Transfer

    Directory of Open Access Journals (Sweden)

    Zhang John

    2016-07-01

    Full Text Available Minimal stimulation in vitro fertilization (mini-IVF consists of a gentle controlled ovarian stimulation that aims to produce a maximum of five to six oocytes. There is a misbelief that mini-IVF severely compromises pregnancy and live birth rates. An appraisal of the literature pertaining to studies on mini-IVF protocols was performed. The advantages of minimal stimulation protocols are reported here with a focus on the use of clomiphene citrate (CC, gonadotropin releasing hormone (GnRH ago- nist trigger for oocyte maturation, and freeze-all embryo strategy. Literature review and the author’s own center data suggest that minimal ovarian stimulation protocols with GnRH agonist trigger and freeze-all embryo strategy along with single embryo transfer produce a reasonable clinical pregnancy and live birth rates in both good and poor responders. Additionally, mini-IVF offers numerous advantages such as: i. Reduction in cost and stress with fewer office visits, needle sticks, and ultrasounds, and ii. Reduction in the incidence of ovarian hyperstimulation syndrome (OHSS. Mini-IVF is re-emerging as a solution for some of the problems associated with conventional IVF, such as OHSS, cost, and patient discomfort.

  20. Mild ovarian stimulation for IVF: 10 years later

    DEFF Research Database (Denmark)

    Fauser, Bart C J M; Nargund, Geeta; Andersen, Anders Nyboe;

    2010-01-01

    Ovarian stimulation to achieve multiple follicle development has been an integral part of IVF treatment. In the context of improved laboratory performance, the need for a large number of oocytes as an integral part of a successful IVF programme may be questioned. The aim of the current debate...... is to summarize the studies performed during the last decade to develop the concept of mild stimulation aiming to obtain fewer than eight oocytes. Here we examine the balance between IVF success and patient discomfort, and complications and cost, and how these might improve by simpler ovarian stimulation...

  1. INVO Procedure: Minimally Invasive IVF as an Alternative Treatment Option for Infertile Couples

    Directory of Open Access Journals (Sweden)

    Elkin Lucena

    2012-01-01

    Full Text Available Intravaginal culture (IVC, also called INVO (intravaginal culture of oocytes, is an assisted reproduction procedure where oocyte fertilization and early embryo development are carried out within a gas permeable air-free plastic device, placed into the maternal vaginal cavity for incubation. In the present study we assessed the outcome of the INVO procedure, using the recently designed INVOcell device, in combination with a mild ovarian stimulation protocol. A total of 125 cycles were performed. On average 6.5 oocytes per cycle were retrieved, and a mean of 4.2 were placed per INVOcell device. The cleavage rate obtained after the INVO culture was 63%. The procedure yielded 40%, 31.2%, and 24% of clinical pregnancy, live birth, and single live birth rates per cycle, respectively. Our results suggest that the INVO procedure is an effective alternative treatment option in assisted reproduction that shows comparable results to those reported for existing IVF techniques.

  2. Relationship between Different Pronuclear Patterns and Potential of Embryo Development and Pregnancy

    Institute of Scientific and Technical Information of China (English)

    Zhong-mei LI; Yu-bao WANG; Dun-yu GU; Xiao-yun WU; De-qi JIANG; Zhen LU; Zuan-chong FENG

    2003-01-01

    Objective To explore the relationship between the patterns of pronucleus and embryo development and pregnancy potential in the pronuclear stageMethods According to the number and distribution of nucleolar precursor bodies, the embryos at pronuclear stage were classified into 6 pronuclear patterns from 0 to 5, 16~18 h after in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). For each pattern, the subsequent embryonic morphology and the pregnancy rate were analyzed. Results Embryos of Pattern 0 developed to significantly more embryos with good quality and higher pregnancy potential than the embryos developing from other patterns (83.14% and 76.11% respectively, P<0.05). The pregnancy rate was decreased as less embryos of Pattern 0 were transferred . The pregnancy rate of the groups of only Pattern 0, with Pattern 0, and without Pattern 0 were 48.08%, 32.14% and 21.28% respectively (P<0.05).Conclusions The pronuclear patterns are of the predictive value of embryo development and pregnancy potential, which can be used as a new tool for the selection of embryos in IVF and ICSI.

  3. 卵巢低反应患者IVF-ET周期中AMH的检测价值及护理对策%Value of Measuring AMH Level and Nursing Strategies in Poor Ovarian Response Patients Undergoing IVF-ET

    Institute of Scientific and Technical Information of China (English)

    杨晓萍; 陈钦洁; 李丽娜; 张军

    2015-01-01

    目的 探讨卵巢低反应患者体外受精-胚胎移植(in vitro fertilization and embryo transfer,IVF-ET)周期中血清抗苗勒管激素(anti-mullerian hormone,AMH)检测的价值及护理对策.方法 采用前瞻性研究方法分析南方医院妇产科生殖中心2014年1-12月接受IVF-ET治疗的112例不孕症女性患者的资料,ELISA方法检测血清AMH水平,偏相关方法分析血清AMH水平与IVF结局参数的关系;并行针对性心理疏导、用药宣教、手术护理、健康宣教等护理措施.结果 卵巢低反应患者IVF-ET周期中血清AMH水平与获卵数和优质胚胎数均成正相关.结论 血清AMH水平对预测卵巢低反应者IVF周期卵子和胚胎质量有指导意义,其中血清AMH水平下降提示卵巢低反应者IVF-ET周期卵子和胚胎质量发育欠佳.针对此类患者,应制定相应的护理对策,使患者在整个治疗周期中保持自然轻松的心理状态,积极配合治疗,并坦然接受治疗结果.

  4. Characterisation of bovine epiblast-derived outgrowth colonies

    DEFF Research Database (Denmark)

    Østrup, Esben; Gjørret, Jakob; Schauser, Kirsten Hallundbæk;

    2010-01-01

    The aim of the present study was to characterise bovine epiblast-derived outgrowth colonies (OCs) with respect to the embryonic origin of their cellular components. Epiblasts were isolated mechanically from bovine Day 12 embryos. Epiblasts were cultured on feeder layers of SNL cells (neomycin...

  5. Synergistic Effect of Insulin on in vitro Development of Immature Bovine Oocytes

    OpenAIRE

    Mojtaba Dashtizad; Abd W. Haron; Rosnina Yusoff; Morteza Daliri; Hadi Hajarian; Mehdi Najari; Yap K. Chee; Abas M. Othman

    2010-01-01

    Problem statement: Development of efficient culture system to support embryonic development would be valuable when quality of produced embryos was important. However, the rate of bovine embryo production in vitro was still lower than expected. Present study, including of three experiments, was carried out to investigate the effect of insulin on nuclear maturation and subsequent development of immature bovine oocytes and in vitro fertilized embryos. Approach: Grade one cumulus-oocyte-complexes...

  6. First Irish birth following IVF therapy using antagonist protocol.

    LENUS (Irish Health Repository)

    Mocanu, E V

    2012-02-01

    BACKGROUND: During in vitro fertilization (IVF), the prevention of a premature LH surge was traditionally achieved using a gonadotrophin releasing hormone agonist (GnRH-a), and more recently, a GnRH antagonist. AIMS: We report a case of a 37 year old treated using the GnRH antagonist in a second completed cycle of IVF. METHODS: IVF was performed for primary infertility of 5-year duration due to frozen pelvis secondary to endometriosis. RESULTS: Following controlled ovarian hyperstimulation, oocyte recovery and fertilization, cleavage and transfer of two zygotes, a pregnancy established. A twin gestation was diagnosed at 7-weeks scan and pregnancy ended with the delivery of twin girls by emergency caesarean section. CONCLUSION: This is a first report of a delivery following IVF using the antagonist protocol in Ireland. Such therapy is patient friendly and its use should be introduced on a larger scale in clinical practice.

  7. Cost-effectiveness of primary offer of IVF vs. primary offer of IUI followed by IVF (for IUI failures) in couples with unexplained or mild male factor subfertility

    OpenAIRE

    Pashayan, Nora; Lyratzopoulos, Georgios; Mathur, Raj

    2006-01-01

    Background In unexplained and mild male factor subfertility, both intrauterine insemination (IUI) and in-vitro fertilisation (IVF) are indicated as first line treatments. Because the success rate of IUI is low, many couples failing IUI subsequently require IVF treatment. In practice, it is therefore important to examine the comparative outcomes (live birth-producing pregnancy), costs, and cost-effectiveness of primary offer of IVF, compared with primary offer of IUI followed by IVF for couple...

  8. Cost-effectiveness of primary offer of IVF vs. primary offer of IUI followed by IVF (for IUI failures) in couples with unexplained or mild male factor subfertility

    OpenAIRE

    Pashayan, N.; Lyratzopoulos, G.; Mathur, R

    2006-01-01

    Background: In unexplained and mild male factor subfertility, both intrauterine insemination (IUI) and in-vitro fertilisation (IVF) are indicated as first line treatments. Because the success rate of IUI is low, many couples failing IUI subsequently require IVF treatment. In practice, it is therefore important to examine the comparative outcomes ( live birth-producing pregnancy), costs, and cost-effectiveness of primary offer of IVF, compared with primary offer of IUI followed by IVF for coup...

  9. Cost-effectiveness of primary offer of IVF vs. primary offer of IUI followed by IVF (for IUI failures) in couples with unexplained or mild male factor subfertility

    OpenAIRE

    Lyratzopoulos Georgios; Pashayan Nora; Mathur Raj

    2006-01-01

    Abstract Background In unexplained and mild male factor subfertility, both intrauterine insemination (IUI) and in-vitro fertilisation (IVF) are indicated as first line treatments. Because the success rate of IUI is low, many couples failing IUI subsequently require IVF treatment. In practice, it is therefore important to examine the comparative outcomes (live birth-producing pregnancy), costs, and cost-effectiveness of primary offer of IVF, compared with primary offer of IUI followed by IVF f...

  10. Cost-effectiveness of primary offer of IVF vs. primary offer of IUI followed by IVF (for IUI failures in couples with unexplained or mild male factor subfertility

    Directory of Open Access Journals (Sweden)

    Lyratzopoulos Georgios

    2006-06-01

    Full Text Available Abstract Background In unexplained and mild male factor subfertility, both intrauterine insemination (IUI and in-vitro fertilisation (IVF are indicated as first line treatments. Because the success rate of IUI is low, many couples failing IUI subsequently require IVF treatment. In practice, it is therefore important to examine the comparative outcomes (live birth-producing pregnancy, costs, and cost-effectiveness of primary offer of IVF, compared with primary offer of IUI followed by IVF for couples failing IUI. Methods Mathematical modelling was used to estimate comparative clinical and cost effectiveness of either primary offer of one full IVF cycle (including frozen cycles when applicable or "IUI + IVF" (defined as primary IUI followed by IVF for IUI failures to a hypothetical cohort of subfertile couples who are eligible for both treatment strategies. Data used in calculations were derived from the published peer-reviewed literature as well as activity data of local infertility units. Results Cost-effectiveness ratios for IVF, "unstimulated-IUI (U-IUI + IVF", and "stimulated IUI (S-IUI + IVF" were £12,600, £13,100 and £15,100 per live birth-producing pregnancy respectively. For a hypothetical cohort of 100 couples with unexplained or mild male factor subfertility, compared with primary offer of IVF, 6 cycles of "U-IUI + IVF" or of "S-IUI + IVF" would cost an additional £174,200 and £438,000, representing an opportunity cost of 54 and 136 additional IVF cycles and 14 to 35 live birth-producing pregnancies respectively. Conclusion For couples with unexplained and mild male factor subfertility, primary offer of a full IVF cycle is less costly and more cost-effective than providing IUI (of any modality followed by IVF.

  11. The inSIGHT study: costs and effects of routine hysteroscopy prior to a first IVF treatment cycle. A randomised controlled trial

    Directory of Open Access Journals (Sweden)

    Smit Janine G

    2012-08-01

    Full Text Available Abstract Background In in vitro fertilization (IVF and intracytoplasmatic sperm injection (ICSI treatment a large drop is present between embryo transfer and occurrence of pregnancy. The implantation rate per embryo transferred is only 30%. Studies have shown that minor intrauterine abnormalities can be found in 11–45% of infertile women with a normal transvaginal sonography or hysterosalpingography. Two randomised controlled trials have indicated that detection and treatment of these abnormalities by office hysteroscopy after two failed IVF cycles leads to a 9–13% increase in pregnancy rate. Therefore, screening of all infertile women for intracavitary pathology prior to the start of IVF/ICSI is increasingly advocated. In absence of a scientific basis for such a policy, this study will assess the effects and costs of screening for and treatment of unsuspected intrauterine abnormalities by routine office hysteroscopy, with or without saline infusion sonography (SIS, prior to a first IVF/ICSI cycle. Methods/design Multicenter randomised controlled trial in asymptomatic subfertile women, indicated for a first IVF/ICSI treatment cycle, with normal findings at transvaginal sonography. Women with recurrent miscarriages, prior hysteroscopy treatment and intermenstrual blood loss will not be included. Participants will be randomised for a routine fertility work-up with additional (SIS and hysteroscopy with on-the-spot-treatment of predefined intrauterine abnormalities versus the regular fertility work-up without additional diagnostic tests. The primary study outcome is the cumulative ongoing pregnancy rate resulting in live birth achieved within 18 months of IVF/ICSI treatment after randomisation. Secondary study outcome parameters are the cumulative implantation rate; cumulative miscarriage rate; patient preference and patient tolerance of a SIS and hysteroscopy procedure. All data will be analysed according to the intention-to-treat principle

  12. The Relationship between Cell Number, Division Behavior and Developmental Potential of Cleavage Stage Human Embryos: A Time-Lapse Study.

    Science.gov (United States)

    Kong, Xiangyi; Yang, Shuting; Gong, Fei; Lu, Changfu; Zhang, Shuoping; Lu, Guangxiu; Lin, Ge

    2016-01-01

    Day 3 cleavage embryo transfer is routine in many assisted reproductive technology centers today. Embryos are usually selected according to cell number, cell symmetry and fragmentation for transfer. Many studies have showed the relationship between cell number and embryo developmental potential. However, there is limited understanding of embryo division behavior and their association with embryo cell number and developmental potential. A retrospective and observational study was conducted to investigate how different division behaviors affect cell number and developmental potential of day 3 embryos by time-lapse imaging. Based on cell number at day 3, the embryos (from 104 IVF/intracytoplasmic sperm injection (ICSI) treatment cycles, n = 799) were classified as follows: less than 5 cells (10C; n = 42). Division behavior, morphokinetic parameters and blastocyst formation rate were analyzed in 5 groups of day 3 embryos with different cell numbers. In 10C embryos increased compared to 7-8C embryos (45.8%, 33.3% vs. 11.1%, respectively). In ≥5C embryos, FR and DC significantly reduced developmental potential, whereas division behaviors. In NB embryos, the blastocyst formation rate increased with cell number from 7.4% (10C). In NB embryos, the cell cycle elongation or shortening was the main cause for abnormally low or high cell number, respectively. After excluding embryos with abnormal division behaviors, the developmental potential, implantation rate and live birth rate of day 3 embryos increased with cell number.

  13. THE PREDICTORS OF CLINICAL FERTILIZATION IN IVF CYCLES

    OpenAIRE

    S. Esmailzadeh; M. Faramarzi M. Farsi

    2008-01-01

    We examined the demographic characteristics of couples, ovarian response and sperm quality to determine the predictors of clinical fertilization in IVF cycles. A descriptive and analytic study was conducted using data and biologic specimens obtained Fatemeh-Alzahra of Babol infertility center from 2004 to 2005. Only data from 315 women who had medical indication for conventional IVF treatment were included in the analysis. Treatment using ICSI was excluded. In the univariate analysis, the fol...

  14. SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

    Institute of Scientific and Technical Information of China (English)

    Fang-yin Meng; Xiao-hong Li

    2004-01-01

    @@ Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.

  15. Risks of In Vitro Fertilization (IVF)

    Science.gov (United States)

    ... see the ASRM booklet titled Multiple pregnancy and birth: twins, triplets and high- order multiples . One way to avoid multiple pregnancy is to choose to transfer only one embryo at a time. For more information about this, ... increase the risk of my child having a birth defect? The risk of birth defects in the ...

  16. IVF-ET术后流产患者绒毛和蜕膜组织凋亡及相关调控蛋白的表达%Human chorionic villi and deciduas apoptosis and related modulin expression of IVF-ET failure

    Institute of Scientific and Technical Information of China (English)

    张先霞; 王繁

    2012-01-01

    目的:检测体外受精-胚胎移植(IVF-ET)术后早期自然流产患者绒毛和蜕膜组织凋亡及相关调控蛋白血管内皮生长因子(VEGF)、Bcl-2的表达,探讨其发病和基因调控机制.方法:采用免疫组织化学染色技术检测IVF-ET术后早期自然流产者(IVF-ET组)绒毛及蜕膜组织Bcl-2、VEGF的表达水平,并以同期正常早孕者(正常早孕组)的标本为对照.结果:VEGF主要表达于绒毛滋养细胞和血管内皮细胞胞浆.IVF-ET组VEGF表达明显低于正常早孕组,差异具有统计学意义(P<0.05).Bcl-2主要在绒毛合体滋养细胞上表达.IVF-ET组滋养细胞及蜕膜细胞染色较浅,表达呈弱阳性,正常早孕组染色较深,表达强阳性,差异有统计学意义(P<0.05).结论:IVF-ET术后早期自然流产者绒毛及蜕膜组织中VEGF及Bcl-2表达显著降低,提示早期妊娠中VEGF及Bcl-2的低表达可能是导致IVF-ET术后早期自然流产的原因之一.%Objective: To investigate the human chorionic villi and deciduas apoptosis and related regulatory protein (VEGF, Bc1-2) expression in early spontaneous abortion after treatment of in vitro fertilization-embryo transfer (IVF-ET) and explore the pathomechanisra of spontaneous abortion. Methods: The expression of VEGF and Bcl-2 in the chorionic villi and deciduas was detected and analyzed in 30 cases of early spontaneous abortion patients after in vitro fertilization-embryo transfer (IVF-ET) through immunohistochemisty method, Thirty cases of normal early pregnancy as control. Results: Immunohistochemistry displayed that VEGF mainly expressed in cytoplasm of villi trophocyte and vascular endothelial cell. The VEGF level of early spontaneous abortion after IVF-ET group was significantly lower than that of normal early pregnancy group (P<0.05). Bcl-2 mainly expressed in the villi syncytiotrophoblast. Trophoblasts and decidual staining were shallow and weakly positive in early spontaneous abortion after IVF-ET group. But

  17. Comparison between a GnRH Agonist and a GnRH Antagonist Protocol for the Same Patient Undergoing IVF

    Institute of Scientific and Technical Information of China (English)

    Yufeng LI; Yuan LI; Qiaohong LAI; Hanwang ZHANG; Guijin ZHU; Lei JIN; Jing YUE

    2008-01-01

    Summary: In order to compare GnRH agonist with antagonist protocol for the same patient during controlled ovarian stimulation cycles, the in vitro fertilization and embryo transfer (IVF-ET) outcome was retrospectively studied in 81 patients undergoing 105 agonist protocols and 88 antagonist protocols. The results showed that there was no statistically significant difference in duration of ovarian stimulation, number of ampoules, oocytes retrieved, serum estradiol (E2) and progesterone (P) levels,thickness of endometrium, the zygote-and blastocyst-developmcnt rate between GnRH agonist and antagonist protocols (P>0.05). High quality embryo rate was higher in antagonist protocols, but there was no significant difference between two protocols. Implantation rate and clinical pregnant rate were significantly higher in antagonist protocol (15.82% and 30.26%, respectively) than in agonist protocol (5.26% and 10.64% respectively (P<0.05). It was concluded GnRH antagonist protocol probably improved the outcome of pregnancy of older patients with a history of multiple failure of IVF-ET in a GnRH protocol.

  18. Analysis of Abortion during IVF-ET Treatment%IVF-ET过程中流产因素分析

    Institute of Scientific and Technical Information of China (English)

    王俊超; 徐凤琴; 罗海宁; 马俊芳; 张云山

    2012-01-01

    目的:探讨在体外受精-胚胎移植(TVF-ET)过程中引起患者流产的因素.方法:选择2005年1月-2009年12月行IVF-ET且成功临床妊娠的患者1195例,其中流产患者203例,包括早期流产患者150例,为A组;晚期流产患者53例,为B组;持续妊娠及分娩患者992例,为C组.分别对3组患者年龄、体质量指数(BMI)、超排卵(COH)情况、临床妊娠情况及是否行减胎等治疗进行比较和分析.结果:A组患者年龄、促性腺激素(Gn)总量明显高于C组;B组BMI明显高于A组、C组.A组ET后第14天β-hCG水平明显低于B组、C组而单胎妊娠率明显高于B、C两组,B组胎儿畸形率、减胎率明显高于A组、C组.3组患者注射hCG日发育卵泡数、获卵数、雌二醇水平及受精方式差异无统计学意义.结论:高BMI、大剂量Gn超排卵以及多胎妊娠和减胎术是IVF-ET时造成流产的重要因素,ET后14d血β-hCG水平偏低提示胚胎发育不良、早期流产的风险大.%Objective:Methods: Two hundred and three patients including ISO patients with early abortion (group A) and S3 patients with late abortion (group B) were enrolled, while 992 patients with continuing pregnancy and full-term pregnancy were as control (group C). The age, body mass index (BMI), hyperstimulation (COH) and clinical pregnancy results were compared between three groups. Results: The age, total dosage of gonadotropin (Gn) and the single pregnancy rate were significantly higher in group A than those in group B and group C. The value of BMI was higher in group B than that of group A and group C. The level of β-hCG was lower in group A 14 days after ET treatment than that of group B and group C, but the pregnancy rate was significantly higher in group A than that of group B and group C. Rates of fetal malformation and fetal reduction were significantly higher in group B than those in group A and group C. There were no significant differences in numbers of follicle development and

  19. DNA methylation in porcine preimplantation embryos developed in-vivo or produced by in-vitro fertilization, parthenogenetic activation and somatic cell nuclear transfer

    DEFF Research Database (Denmark)

    Deshmukh, Rahul Shahaji; Østrup, Olga; Østrup, Esben;

    2011-01-01

    vitro fertilized (IVF), somatic cell nuclear transfer (SCNT) and parthenogenetically activated (PA) embryos were evaluated for DNA methylation quantification at different developmental stages. Fertilized (IV and IVF) one-cell stages lacked a substantial active demethylation of the paternal genome....... Embryos produced under in vitro conditions had higher levels of DNA methylation than IV. A lineage-specific DNA methylation (hypermethylation of the inner cell mass and hypomethylation of the trophectoderm) was observed in porcine IV late blastocysts, but was absent in PA- and SCNT-derived blastocysts...

  20. Recent Progress in Cryopreservation of Bovine Oocytes

    Directory of Open Access Journals (Sweden)

    In-Sul Hwang

    2014-01-01

    Full Text Available Principle of oocyte cryoinjury is first overviewed and then research history of cryopreservation using bovine oocytes is summarized for the last two decades with a few special references to recent progresses. Various types of cryodevices have been developed to accelerate the cooling rate and applied to the oocytes from large domestic species enriched with cytoplasmic lipid droplets. Two recent approaches include the qualitative improvement of IVM oocytes prior to the vitrification and the short-term recovery culture of vitrified-warmed oocytes prior to the subsequent IVF. Supplementation of L-carnitine to IVM medium of bovine oocytes has been reported to reduce the amount of cytoplasmic lipid droplets and improve the cryotolerance of the oocytes, but it is still controversial whether the positive effect of L-carnitine is reproducible. Incidence of multiple aster formation, a possible cause for low developmental potential of vitrified-warmed bovine oocytes, was inhibited by a short-term culture of the postwarm oocytes in the presence of Rho-associated coiled-coil kinase (ROCK inhibitor. Use of an antioxidant α-tocopherol, instead of the ROCK inhibitor, also supported the revivability of the postwarm bovine oocytes. Further improvements of the vitrification procedure, combined with pre- and postvitrification chemical treatment, would overcome the high sensitivity of bovine oocytes to cryopreservation.

  1. 体细胞核移植技术生产转入溶菌酶基因(hLYZ)牛胚胎的研究%Human Lysozyme Gene (hLYZ) Transgenic Bovine Embryos Produced by Somatic Cell Nuclear Transfer

    Institute of Scientific and Technical Information of China (English)

    熊显荣; 李文哲; 王丽君; 王勇胜; 苏建民; 华松; 张涌

    2011-01-01

    The aim of this study was to investigate the effect of different sizes and treatments of donor cells on the in vitro development of bovine embryos after genetic modification and selection, so as to establish an effective system for production of transgenic bovine embryos.Bovine fetal fibroblasts were transfeected with the recombinant plasmid of mammary gland specific expression vector pEBH which containing human lysozyme (hLYZ) gene.Transgenic positive cells which were obtained through G418 selection were used as donor cells for production of transgenic cloned embryos.The result showed that genetic modification and screening of donor cells were harmful for the development of transgenic cloned embryos, the blastocyst rate decreased significantly;referred to internal diameter of injective needle, cells with the diameter of 15~20 μm were selected and used as nuclear transfer donor cells, the fusion rate and blastocyst rate of transgenic cloned embryos were significant higher than that of the other groups (P < 0.05); Compared with serum starvation and contact restrain groups, the cleavage rate and blastocyst rate of transgenic cloned embryos which were constructed with plant cells were significantly higher (P < 0.05); The expression of GFP was observed in each stage of in vitro development in all transgenic cloned embryos, but the expression levels seemed to be vary among individuals and even among different developmental stages of the same individual.We chose I 0 embroys randomly for PCR detection and found that all of the embroys were positive.Above all, we obtained hLYZ transgenic cloned embryos by somatic cell nuclear transfer technique, the reconstructed embryos can develop to blastocysts successfully; and when plant cells with diameter of 15~20 μm were used as donor cell, the developmental competence of somatic cell cloned embryos is improved significantly.%本研究主要探讨经基因修饰和筛选后转基因供体细胞的挑选及处理对牛转

  2. 认知联合音乐疗法对 IVF - ET 夫妇生活质量的影响%Impact of cognitive therapy combined with music therapy on quality of life for the IVF - ET couples

    Institute of Scientific and Technical Information of China (English)

    刘芳; 王爱华; 王丽; 张林

    2015-01-01

    目的:探讨认知联合音乐疗法对体外受精-胚胎移植(IVF - ET)夫妇生活质量的影响。方法:将2013年12月~2014年5月符合标准的32对 IVF - ET 夫妇随机等分为对照组和试验组。对照组仅给予常规治疗和护理,试验组在此基础上实施认知联合音乐疗法进行干预。干预8周后使用 SF -36量表进行评价。结果:8周后试验组夫妇在生理职能、总体健康状况、精力、社会功能、情感职能、精神健康的得分及生活质量总分均高于对照组,差异有统计学意义(P 均﹤0.05)。结论:认知联合音乐疗法能有效改善 IVF - ET 夫妇躯体健康及心理健康水平,从而提高IVF - ET 夫妇的生活质量。认知疗法联合音乐疗法可以改善 IVF - ET 夫妇双方的生活质量。%Objective:To investigate the effect of cognitive therapy combined with music therapy on quality of life of the couples for the in vitro fertilization embryo transfer(IVF - ET). Methods:32 couples were included in this study. They were randomly divided into control group and experimental group,each group included 16 couples. The control group was given conventional treatment and nursing. The experimental group were also given cognitive therapy com-bined with music therapy on the basis in the conventional treatment and nursing in 8 weeks. Using SF - 36 scale,quality of life were evaluaed respectively on admission a after the 8 weeks of intervention in the two groups. Results:The scores of the physiological function and quality of life were higher in the experi-mental group than those of the control group after intervention,the difference was statistically significant(P ﹤ 0. 05). Conclusion:Cognitive therapy com-bined with music therapy can effectively improve the IVF - ET couple on the physiological function and Somatic function,also can eliminate wives negative emotion,so as to improve the quality of life of IVF - ET couples.

  3. When no choice of embryos exists, the multiple pregnancy risk is still high.

    LENUS (Irish Health Repository)

    Wong, V V

    2012-10-01

    Multiple pregnancies arising from assisted reproduction are known to be associated with increased medical, psychological, economical and social risks. If only two embryos develop after culture, how should the couple be counselled in relation to the risk of multiple pregnancy? We performed a retrospective review of all IVF\\/ICSI treatments performed between 1 January 2005 and 31 December 2007, that resulted in double embryos transfer (DET). We identified 623 cycles with a fortuitous DET (Group I) and 635 cycles with an elective DET (Group II). Group II were significantly more likely to have twins when compared with Group I, irrespective of age. Twin rates in Group I were higher in IVF compared with ICSI; 33.3% vs 16.6% in < 35 years old and 16.2% vs 7.6% in 35-40 years old. Therefore, single embryo transfer should be considered for IVF patients below 35 years old, even if only two good quality embryos are available.

  4. Pregnancy, Delivery, and Neonatal Outcomes of In Vitro Fertilization-Embryo Transfer in Patient with Previous Cesarean Scar.

    Science.gov (United States)

    Zhang, Ningyuan; Chen, Hua; Xu, Zhipeng; Wang, Bin; Sun, Haixiang; Hu, Yali

    2016-01-01

    BACKGROUND What role should previous cesarean section play in affecting clinical pregnancy outcomes and avoiding the complications of in vitro fertilization? In this article, we focus on elective single-embryo transfer (eSET) versus double-embryo transfer (DET) and assess the clinical efficacy and safety of eSET in patients who have a previous cesarean scar. MATERIAL AND METHODS The pregnancy, delivery, and neonatal outcomes of 130 patients who had a previous cesarean scar and received in vitro fertilization-embryo transfer (IVF-ET) were retrospectively analyzed. The number of transferred embryos was chosen depending on patients' desire after acknowledging all benefits and risks, including eSET (eSET group, n=56) and DET (DET group, n=74). A total of 101 patients with previous vaginal delivery receiving IVF-ET in the same period were included as a control group. RESULTS The pregnancy rates, multiple birth rates, abortion rates, ectopic pregnancy rates, gestational age at delivery, preterm birth rates, neonatal birth weight, and take-home baby rates were similar between the previous cesarean section group and the previous vaginal delivery group. A previous cesarean section scar did not affect embryo implantation and pregnancy outcomes in IVF. In the eSET and DET groups of previous cesarean section patients, the embryo implantation rates, pregnancy rates, abortion rates, and take-home baby rates were similar. However, the rate of multiple pregnancies reached 50% in the DET group, which led to more preterm births and lower birth weight. CONCLUSIONS Elective single-embryo transfer is a well-accepted strategy to avoid multiple pregnancies and improve the obstetric and neonatal outcomes of singleton pregnancy in IVF patients with a previous cesarean section. PMID:27636504

  5. Placental development during early pregnancy in sheep: Effects of embryo origin on vascularization

    Science.gov (United States)

    Grazul-Bilska, Anna T.; Johnson, Mary Lynn; Borowicz, Pawel P.; Bilski, Jerzy J.; Cymbaluk, Taylor; Norberg, Spencer; Redmer, Dale A.; Reynolds, Lawrence P.

    2014-01-01

    Utero-placental growth and vascular development are critical for pregnancy establishment that may be altered by various factors including assisted reproductive technologies (ART), nutrition, or others, leading to compromised pregnancy. We hypothesized that placental vascularization and expression of angiogenic factors are altered early in pregnancies after transfer of embryos created using selected ART methods. Pregnancies were achieved through natural mating (NAT), or transfer of embryos from natural mating (NAT-ET), or in vitro fertilization (IVF) or activation (IVA). Placental tissues were collected on day 22 of pregnancy. In maternal caruncles (CAR), vascular cell proliferation was less (P<0.05) for IVA than other groups. Compared to NAT, density of blood vessels was less (P<0.05) for IVF and IVA in fetal membranes (FM), and for NAT-ET, IVF and IVA in CAR. In FM, mRNA expression was decreased (P<0.01–0.08) in NAT-ET, IVF and IVA compared to NAT for vascular endothelial growth factor (VEGF) and its receptor FLT-1, placental growth factor (PGF), neuropilin (NP) 1 and 2, angiopoietin (ANGPT) 1 and 2, endothelial nitric oxide synthase (NOS3), hypoxia inducible factor-1A (HIF1A), fibroblast growth factor (FGF) 2 and its receptor FGFR2. In CAR, mRNA expression was decreased (P<0.01–0.05) in NAT-ET, IVF and IVA compared to NAT for VEGF, FLT-1, PGF, ANGPT1 and TEK. Decreased mRNA expression for 12 of 14 angiogenic factors across FM and CAR in NAT-ET, IVF and IVA pregnancies was associated with reduced placental vascular development, which would lead to poor placental function and compromised fetal and placental growth and development. PMID:24472816

  6. Embryo quality predictive models based on cumulus cells gene expression

    Directory of Open Access Journals (Sweden)

    Devjak R

    2016-07-01

    Full Text Available Since the introduction of in vitro fertilization (IVF in clinical practice of infertility treatment, the indicators for high quality embryos were investigated. Cumulus cells (CC have a specific gene expression profile according to the developmental potential of the oocyte they are surrounding, and therefore, specific gene expression could be used as a biomarker. The aim of our study was to combine more than one biomarker to observe improvement in prediction value of embryo development. In this study, 58 CC samples from 17 IVF patients were analyzed. This study was approved by the Republic of Slovenia National Medical Ethics Committee. Gene expression analysis [quantitative real time polymerase chain reaction (qPCR] for five genes, analyzed according to embryo quality level, was performed. Two prediction models were tested for embryo quality prediction: a binary logistic and a decision tree model. As the main outcome, gene expression levels for five genes were taken and the area under the curve (AUC for two prediction models were calculated. Among tested genes, AMHR2 and LIF showed significant expression difference between high quality and low quality embryos. These two genes were used for the construction of two prediction models: the binary logistic model yielded an AUC of 0.72 ± 0.08 and the decision tree model yielded an AUC of 0.73 ± 0.03. Two different prediction models yielded similar predictive power to differentiate high and low quality embryos. In terms of eventual clinical decision making, the decision tree model resulted in easy-to-interpret rules that are highly applicable in clinical practice.

  7. Why do couples discontinue unlimited free IVF treatments?

    Science.gov (United States)

    Lande, Yechezkel; Seidman, Daniel S; Maman, Ettie; Baum, Micha; Hourvitz, Ariel

    2015-03-01

    Worldwide, IVF is often discontinued before a live birth is achieved due to high costs. Even when partial financial coverage is provided, often medical providers advise treatment discontinuation. In Israel, unlimited IVF is offered free of charge for a couples' first two children. Our objective was to assess the reasons couples discontinue IVF treatments before achieving two children in a completely unlimited cost-free environment. This cohort study included all primary infertile women, reason they ceased treatments. Of the 134 couples included, only 46 ceased IVF treatments without achieving two children, after performing an average of 6.2 IVF cycles to achieve their first birth. The reasons given were: lost hope of success (13), psychological burden (18), divorce (6), medical staff recommendation (5), bureaucratic difficulties (3) and general medical condition (1). The main reasons for "drop out" in our cost-free environment were as follows: psychological burden and lost hope of success. Due to high availability of treatments, medical staff recommendation was a less significant factor in our study.

  8. Ovarian endometriomas and IVF: a retrospective case-control study

    Directory of Open Access Journals (Sweden)

    Guidetti Daniela

    2011-06-01

    Full Text Available Abstract We performed this retrospective case-control study analyzing 428 first-attempt in vitro fertilization (IVF cycles, among which 254 involved women with a previous or present diagnosis of ovarian endometriosis. First, the results of these 254 cycles were compared with 174 cycles involving patients with proven non-endometriotic tubal infertility having similar age and body mass index. Women with ovarian endometriosis had a significantly higher cancellation rate, but similar pregnancy, implantation and delivery rates as patients with tubal infertility. Second, among the women with ovarian endometriosis, the women with a history of laparoscopic surgery for ovarian endometriomas prior to IVF and no visual endometriosis at ovum pick-up (n = 112 were compared with the non-operated women and visual endometriomas at ovum pick-up (n = 142. Patients who underwent ovarian surgery before IVF had significantly shorter period, lower antral follicle count and required higher gonadotropin doses than patients with non-operated endometriomas. The two groups of women with a previous or present ovarian endometriosis did, however, have similar pregnancy, implantation and live birth rates. In conclusion, ovarian endometriosis does not reduce IVF outcome compared with tubal factor. Furthermore, laparoscopic removal of endometriomas does not improve IVF results, but may cause a decrease of ovarian responsiveness to gonadotropins.

  9. Follicular-fluid anti-Mullerian hormone (FF AMH is a plausible biochemical indicator of functional viability of oocyte in conventional in vitro fertilization (IVF cycles

    Directory of Open Access Journals (Sweden)

    Bindu N Mehta

    2013-01-01

    Full Text Available Context: Oocyte quality may be a governing factor in influencing in vitro fertilization (IVF outcomes. However, morphological evaluation of oocyte quality is difficult in conventional IVF cycles. Follicular-fluid (FF, the site for oocyte growth and development, has not yet been sufficiently explored to obtain a marker indicative of oocyte quality. Anti-Mullerian hormone (AMH is produced by granulosa cells of preantral and early-antral follicles and is released in FF. Aim: To investigate AMH as a biochemical indicator of functional viability/quality of oocyte produced in the FF micro-environmental milieu. Settings and Design: Prospective study involving 132 cycles of conventional IVF-embryo transfer (ET in infertile women. Subjects and Methods: AMH concentration was estimated in pooled FF on day of oocyte pickup. Cycles were sorted into low and high groups according to median (50 th centile values of measurement. Main outcome measure was oocyte viability, which included morphological assessment of oocyte quality, fertilization rate, clinical pregnancy, and implantation rates. Statistical Analysis: Graph-pad Prism 5 statistical package. Results: Low FF AMH group shows significantly higher percentage of top-quality oocytes (65.08 ± 24.88 vs. 50.18 ± 25.01%, P =0.0126, fertilization (83.65 ± 18.38 vs. 75.78 ± 21.02%, P =0.0171, clinical pregnancy (57.57 vs. 16.67%, P <0.0001, and embryo implantation rates (29.79 vs. 7.69%, P <0.0001 compared to high FF AMH group. FF AMH shares an inverse correlation with FF E2 (Pearson r = −0.43, r 2 = 0.18 and clinical pregnancy (Pearson r = −0.46, r 2 = 0.21. Threshold value of FF AMH for pregnancy is <1.750 ng/mg protein. Conclusion: FF AMH is a plausible biochemical indicator of functional viability of oocyte in conventional IVF cycles.

  10. Outcome of IVF in DES-Exposed Daughters: Experience in the 90s

    OpenAIRE

    Pal, Lubna; Shifren, Jan L; Isaacson, Keith B.; Chang, Yuchiao; Marean, Martha; Leykin, Lucy; Toth, Thomas L.

    1997-01-01

    Purpose: The outcome of in vitro fertilization (IVF) in a group of infertile women with a history of in utero exposure to diethylstilbestrol (DES) was analyzed. Records from an academic IVF program were retrospectively reviewed.

  11. Selection of single blastocysts for fresh transfer via standard morphology assessment alone and with array CGH for good prognosis IVF patients: results from a randomized pilot study

    Directory of Open Access Journals (Sweden)

    Yang Zhihong

    2012-05-01

    Full Text Available Abstract Background Single embryo transfer (SET remains underutilized as a strategy to reduce multiple gestation risk in IVF, and its overall lower pregnancy rate underscores the need for improved techniques to select one embryo for fresh transfer. This study explored use of comprehensive chromosomal screening by array CGH (aCGH to provide this advantage and improve pregnancy rate from SET. Methods First-time IVF patients with a good prognosis (age Results For patients in Group A (n = 55, 425 blastocysts were biopsied and analyzed via aCGH (7.7 blastocysts/patient. Aneuploidy was detected in 191/425 (44.9% of blastocysts in this group. For patients in Group B (n = 48, 389 blastocysts were microscopically examined (8.1 blastocysts/patient. Clinical pregnancy rate was significantly higher in the morphology + aCGH group compared to the morphology-only group (70.9 and 45.8%, respectively; p = 0.017; ongoing pregnancy rate for Groups A and B were 69.1 vs. 41.7%, respectively (p = 0.009. There were no twin pregnancies. Conclusion Although aCGH followed by frozen embryo transfer has been used to screen at risk embryos (e.g., known parental chromosomal translocation or history of recurrent pregnancy loss, this is the first description of aCGH fully integrated with a clinical IVF program to select single blastocysts for fresh SET in good prognosis patients. The observed aneuploidy rate (44.9% among biopsied blastocysts highlights the inherent imprecision of SET when conventional morphology is used alone. Embryos randomized to the aCGH group implanted with greater efficiency, resulted in clinical pregnancy more often, and yielded a lower miscarriage rate than those selected without aCGH. Additional studies are needed to verify our pilot data and confirm a role for on-site, rapid aCGH for IVF patients contemplating fresh SET.

  12. Monozygotic Triplets and Dizygotic Twins following Transfer of Three Poor-Quality Cleavage Stage Embryos

    Directory of Open Access Journals (Sweden)

    Reshef Tal

    2012-01-01

    Full Text Available Background. Assisted reproductive technology has been linked to the increased incidence of monozygotic twinning. It is of clinical importance due to the increased risk of complications in multiple pregnancies in general and in monozygotic twins in particular. Case. A 29-year-old female, nulligravida underwent her first IVF cycle. Three poor-quality cleavage stage embryos were transferred resulting in monochorionic triamniotic triplets and dichorionic diamniotic twins. Selective embryo reduction was performed at 12 weeks leaving dichorionic twins. The patient underwent emergency cesarean section due to preterm labor and nonreassuring fetal heart tracing at 30 weeks of gestation. Conclusion. Our case emphasizes that even embryos with significant morphological abnormalities should be considered viable and the possibility of simultaneous spontaneous embryo splitting must be factored into determining number of embryos to transfer.

  13. Desarrollo de embriones de bovino obtenidos por fecundación in vitro cultivados con células oviductales o medio condicionado y transferidos a hembras receptoras Bovine embryo development produced by in vitro fertilization cultured with oviductal cell or conditioned medium and transfer to recipients

    Directory of Open Access Journals (Sweden)

    M RATTO

    1999-01-01

    (4- or 8-cell embryos was 62,7 % (64/102 for the zygotes co-cultured with oviductal cells and 66,7 % (100/150 for the zygotes cultured in conditioned medium. The development to the morula stage was 17,6 % (18/102 for the zygotes co-cultured with oviductal cells and 13,3 % (20/150 for the zygotes cultured in conditioned medium. A statiscally significant difference was not found in the development of 4, 8-cell embryos or morula. The development of embryos up to the blastocyst stage was 15,7 % (16/102 for the zygotes co-cultured with oviductal cells. Two blastocysts were transferred to the uterine horn ipsilateral to the CL in two recipients by non-surgical embryo transfer. Pregnancy was confirmed by ultrasonography at 42 and 57 days detecting the presence of one conceptus in each animal. This work has shown that in vitro inseminated of bovine oocytes with espermatozoa prepared with modified BO and co-cultured with oviductal cells, can develop to the blastocysts stage, unlike those that were cultured with conditioned medium. Finally, it is important to mention that this is the first communication in Chile of pregnancy after in vitro fertilization of bovine oocytes

  14. Follicular progesterone concentrations and messenger RNA expression of MATER and OCT-4 in immature bovine oocytes as predictors of developmental competence.

    Science.gov (United States)

    Urrego, R; Herrera-Puerta, E; Chavarria, N A; Camargo, O; Wrenzycki, C; Rodriguez-Osorio, N

    2015-04-15

    The ability of bovine embryos to develop to the blastocyst stage and to implant and generate healthy offspring depends greatly on the competence of the oocyte. Oocyte competence is attributed to its close communication with the follicular environment and to its capacity to synthesize and store substantial amounts of messenger RNA. Higher developmental competence of bovine oocytes has been associated with both the expression of a cohort of developmental genes and the concentration of sex steroids in the follicular fluid. The aim of this study was to identify differences in the expression of FST in cumulus cells and OCT-4 and MATER in oocytes and the influence of the follicular progesterone and follicular estrogen concentration on the competence of bovine oocytes retrieved 30 minutes or 4 hours after slaughter. Cumulus-oocyte complexes (COCs) were left in postmortem ovaries for 30 minutes (group I) or 4 hours (group II) at 30 °C. Aspirated oocytes were then subjected to IVM, IVF, and IVC or were evaluated for MATER and OCT-4 messenger RNA abundance by quantitative real-time polymerase chain reaction. Total RNA was isolated from pools of 100 oocytes for each experimental replicate. Progesterone and estradiol concentration in follicular fluid was evaluated by immunoassay using an IMMULITE 2000 analyzer. Three repeats of in vitro embryo production were performed with a total of 455 (group I) and 470 (group II) COCs. There were no significant differences between the cleavage rates (72 hours postinsemination [hpi]) between both groups (63.5% vs. 69.1%). However, blastocyst (168 hpi) and hatching (216 hpi) rates were higher (P < 0.05) in group II compared with those of group I (21.3% vs. 30.7% and 27.6% vs. 51.5%, respectively). Group II oocytes exhibited the highest MATER and OCT-4 abundance (P < 0.05). Follicular estradiol concentration was not different between both the groups, whereas the progesterone concentration was lower (P ≤ 0.05) in group II follicles. These

  15. Current Status of Comprehensive Chromosome Screening for Elective Single-Embryo Transfer

    Directory of Open Access Journals (Sweden)

    Ming-Yih Wu

    2014-01-01

    Full Text Available Most in vitro fertilization (IVF experts and infertility patients agree that the most ideal assisted reproductive technology (ART outcome is to have a healthy, full-term singleton born. To this end, the most reliable policy is the single-embryo transfer (SET. However, unsatisfactory results in IVF may result from plenty of factors, in which aneuploidy associated with advanced maternal age is a major hurdle. Throughout the past few years, we have got a big leap in advancement of the genetic screening of embryos on aneuploidy, translocation, or mutations. This facilitates a higher success rate in IVF accompanied by the policy of elective SET (eSET. As the cost is lowering while the scale of genome characterization continues to be up over the recent years, the contemporary technologies on trophectoderm biopsy and freezing-thaw, comprehensive chromosome screening (CCS with eSET appear to be getting more and more popular for modern IVF centers. Furthermore, evidence has showen that, by these avant-garde techniques (trophectoderm biopsy, vitrification, and CCS, older infertile women with the help of eSET may have an opportunity to increase the success of their live birth rates approaching those reported in younger infertility patients.

  16. Pituitary down-regulation in IVF/ICSI: consequences for treatment regimens

    NARCIS (Netherlands)

    M.H. Mochtar

    2011-01-01

    Al in 1944 vond de eerste bevruchting plaats van een humane eicel, buiten het lichaam. De IVF (in vitro fertilisatie) was geboren. Er was toen nog een lange weg te gaan naar de geboorte van het eerste echte IVF-kind, Louise Brown, in 1978. Sindsdien zijn er veel onderzoeken uitgevoerd om de IVF-cycl

  17. Evaluation and assessment of semen for IVF/ICSI

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Evaluation and assessment of sement is very important for both diagnosis of male infertility and selection of patients for treatment with IVF or ICSI.In standard IVF,sperm function is essential for normal fertilization:sperm must be able to bind to zona pellucida (ZP),undergo the acrosome reaction and penetrate the ZP and fuse with the oolemma before fertilization takes place.In contrast,most sperm functions are not required for fertilization in ICSI since sperm bypass the ZP and oolemma by injection of a single sperm directly into cytoplasm of oocyte.Therefore,the clinical decision on treatment of patients with either IVF or ICSI is mostly dependent on results of sperm tests.However,conventional semen analyses do not provide accurate information about sperm fertilizing ability since many patients with subtle sperm defects can not be detected.More advanced sperm function tests are required to detect sperm defects that may lead to failure of fertilization in standard IVF.In the last 15years wer performed extensive studies on relationship between sperm functions and fertilization rates by logistic regression analysis in large numbers of IVF patients including 370 patients with zero fertilization rate by IVF.We confirmed sperm morphology assessed strictly was strongly related to fertilisation rate with standard IVF.Thus sperm morphology assessment is very useful for selection of patients for ICSI.We also developed a number of new tests including sperm-ZP binding,sperm-ZP penetration and the ZP-induced AR and evaluated the clinical value of these tests.Sperm-ZP binding and sperm-ZP penetration tests are the most powerful indicators for sperm fertilizing ability in vitro.The ZP-induced AR is highly correlated with sperm-ZP peretration.We discovered a condition we call disordered ZP-induced AR which causes serve infertility in up to 25% men with otherwise idiopathic infertility In conclusion,the combination of semen analysis with advanced sperm function tests provide

  18. Mild ovarian stimulation for IVF: 10 years later

    DEFF Research Database (Denmark)

    Fauser, Bart C J M; Nargund, Geeta; Andersen, Anders Nyboe;

    2010-01-01

    Ovarian stimulation to achieve multiple follicle development has been an integral part of IVF treatment. In the context of improved laboratory performance, the need for a large number of oocytes as an integral part of a successful IVF programme may be questioned. The aim of the current debate is ...... protocols aimed at retrieving fewer oocytes. We intend to analyse why progress has been rather slow and why there is much resistance to mild stimulation. Finally, presumed useful directions for future research will be discussed....

  19. THE EFFECT OF DIFFERENT GONADOTROPIC HORMONES FOR OVARIAN STIMULATION ON IVF TREATMENT OUTCOME%不同促性腺激素促排卵对IVF-ET结局的影响

    Institute of Scientific and Technical Information of China (English)

    赵频; 祁秀娟; 汤秀明; 刘海宁; 刘建新; 刘玉杰; 景秀菊

    2012-01-01

    Objective To compare the outcome between recombinant follicle stimulating hormone (r-FSH) and urinary follicle stimulating hormone (u-FSH) for in vitro fertilization and embryo transfer (IVF-ET) cycles. Methods A retrospective analysis was done for medical records of 173 patients who had undergone IVF-ET treatment following down regulation with a long midluteal phase protocol were divided into r-FSH group (69 cases) and u-FSH group (104 cases). The outcomes of the two groups were compared. Results The dosages of Gn used in r-FSH group were significantly lower than that in u-FSH group (t= - 4. 017,P0. 05). Serum estradiol (E2), luteinizing hormone (LH) and progesterone (P) levels on HCG day were not significantly different (t= - 0. 539 - 0. 938,P>0. 05). There was no significant difference in the number of follicle ≥ 14 mm on HCG day, oocytes retrieved, as well as fine quality embryo,(t=-0. 255 - 1. 778,P>0. 05). There was also no significant difference between the two groups in terms of nidation, pregnancy rate and abortion rate (x2=0. 138 - 3. 142,P> 0.05). Conclusion The treatment outcome of using r-FSH or u-FSH for IVF-ET is similar, but the cost of u-FSH is dramatically lower.%目的 比较基因重组卵泡刺激素(r-FSH)及尿促卵泡刺激素(u-FSH)对体外受精-胚胎移植(IVF-ET)结局的影响.方法 回顾性分析IVF-ET黄体中期长方案降调超促排卵173个周期的临床资料,其中应用r-FSH 69个周期,u-FSH 104个周期.比较两组促性腺激素(Gn)用量、药物费用、应用天数,注射人绒毛膜促性腺激素( HCG)日雌二醇(E2)、黄体生成素(LH)、孕激素(P)水平,以及≥14 mm卵泡数、获卵数、优胚数、着床率、妊娠率及流产率等.结果 r-FSH组Gn用量显著少于u-FSH组(t=-4.017,P<0.001),r-FSH组Gn费用明显高于u-FSH组(t=9.282,P<0.001),但Gn应用天数两组间差异无显著性(t=0.920,P>0.05).两组注射HCG日E2、LH及P水平差异无显著性(t=-0.539~0.938,P>0.05)

  20. GnRH agonist trigger versus hCG trigger in GnRH antagonist in IVF/ICSI cycles: A review article

    Science.gov (United States)

    Alyasin, Ashraf; Mehdinejadiani, Shayesteh; Ghasemi, Marzieh

    2016-01-01

    Routinely, a bolus of 5.000-10.000 IU human chorionic gonadotropin (hCG) is used for the final follicular maturation and ovulation as a standard method. HCG has the same effect of luteinizing hormone (LH) with long half-life. It has the long lutheotrophic effect which increases the risk of ovarian hyper stimulation syndrome (OHSS). Recently, gonadotropin-releasing hormone agonist (GnRH-a) trigger has been used for the induction of final follicular maturation and ovulation with the aim of reducing the OHSS risk. Several studies have shown that the releases of endogenous follicular stimulating hormone (FSH) and LH after administration of GnRH agonist in in vitro fertilization (IVF) cycles are able to precede the final follicular maturation leading to removal of fertile oocyte with normal development of the embryo and ultimately pregnancy. But based on the results of some studies, using GnRH-a trigger leads to defect luteal-phase resulting to reduce the implantation and clinical pregnancy rates and also increase abortion in fresh embryo transfer cycles compared to routine IVF cycle with hCG triggering . Also, in recent years, studies have continued to modify the luteal phase support, so that the fresh embryo transfer is possible too. In this review, we examined the benefits, problems, and also ways to reform GnRH agonist triggering complications. PMID:27738657

  1. Does Unilateral Oocyte Retrieval due to Transvaginally Inaccessible Ovaries, Contrary to Common Beliefs, Affect IVF/ICSI Treatment Outcomes That Much?

    Science.gov (United States)

    Olgan, Safak; Mumusoglu, Sezcan; Bozdag, Gurkan

    2016-01-01

    Objective. To investigate in vitro fertilization (IVF) treatment outcomes of unilateral oocyte retrieval in patients with transvaginally inaccessible ovaries. Study Design. Ninety-two women who underwent unilateral oocyte retrieval were retrospectively matched for age, antral follicle count, and body mass index with 184 women who underwent bilateral oocyte retrieval. Each patient in bilateral oocyte retrieval group had the same number of cumulus oophorus complexes (COCs) from single ovary and had comparable number of follicles (±2) on contralateral site where follicular aspiration was performed. Results. The number of COCs, metaphase-2 oocytes, 2-pronuclei, and top-quality embryos was significantly lower in unilateral oocyte retrieval group. However, proportion of patients with an embryo transfer of at least one top-quality embryo was found to be comparable between unilateral and bilateral oocyte retrieval. Subsequently, clinical pregnancy and live birth rates were found to be similar between the groups. The ROC curve analysis revealed (AUC = 0.74, 95% CI 0.63–0.86, p = 0.001) that retrieved COCs ≥ 5 from single ovary had sensitivity of 76.0% and specificity of 64.2% for occurrence of a clinical pregnancy. Conclusion. The patients with unilateral oocyte retrieval have reasonable chance of success with IVF. The retrieval of ≥5 COCs from accessible ovary might result in better treatment outcomes among these patients. PMID:27123444

  2. Does Unilateral Oocyte Retrieval due to Transvaginally Inaccessible Ovaries, Contrary to Common Beliefs, Affect IVF/ICSI Treatment Outcomes That Much?

    Directory of Open Access Journals (Sweden)

    Safak Olgan

    2016-01-01

    Full Text Available Objective. To investigate in vitro fertilization (IVF treatment outcomes of unilateral oocyte retrieval in patients with transvaginally inaccessible ovaries. Study Design. Ninety-two women who underwent unilateral oocyte retrieval were retrospectively matched for age, antral follicle count, and body mass index with 184 women who underwent bilateral oocyte retrieval. Each patient in bilateral oocyte retrieval group had the same number of cumulus oophorus complexes (COCs from single ovary and had comparable number of follicles (±2 on contralateral site where follicular aspiration was performed. Results. The number of COCs, metaphase-2 oocytes, 2-pronuclei, and top-quality embryos was significantly lower in unilateral oocyte retrieval group. However, proportion of patients with an embryo transfer of at least one top-quality embryo was found to be comparable between unilateral and bilateral oocyte retrieval. Subsequently, clinical pregnancy and live birth rates were found to be similar between the groups. The ROC curve analysis revealed (AUC = 0.74, 95% CI 0.63–0.86, p=0.001 that retrieved COCs ≥ 5 from single ovary had sensitivity of 76.0% and specificity of 64.2% for occurrence of a clinical pregnancy. Conclusion. The patients with unilateral oocyte retrieval have reasonable chance of success with IVF. The retrieval of ≥5 COCs from accessible ovary might result in better treatment outcomes among these patients.

  3. DNA methylation in porcine preimplantation embryos developed in vivo or produced by in vitro fertilization, parthenogenetic activation and somatic cell nuclear transfer

    DEFF Research Database (Denmark)

    Deshmukh, Rahul Shahaji; Østrup, Olga; Østrup, Esben;

    2011-01-01

    ), in vitro fertilized (IVF), somatic cell nuclear transfer (SCNT) and parthenogenetically activated (PA) embryos were evaluated for DNA methylation quantification at different developmental stages. Fertilized (IV and IVF) one-cell stages lacked a substantial active demethylation of the paternal genome....... Embryos produced under in vitro conditions had higher levels of DNA methylation than IV. A lineage-specific DNA methylation (hypermethylation of the inner cell mass and hypomethylation of the trophectoderm) was observed in porcine IV late blastocysts, but was absent in PA- and SCNT-derived blastocysts...... despite the occurrence of de novo methylation in early blastocysts. Comparable levels of DNA methylation were found in IV embryos and in 50% and 14% of SCNT early and late blastocysts, respectively. In conclusion, DNA methylation patterns were adversely affected by in vitro embryo production...

  4. Superovulatory response and embryonic progressive in Iranian Qezel ewes treated with two different concentrations of bovine somatotropin

    Directory of Open Access Journals (Sweden)

    Amir Hossein Asgari Safdar

    2016-05-01

    Conclusions: The treatment 50 mg bovine somatotropins enhance the ratio and growth of the transferable embryos. Embryos of bST-50 treatment indicated an improved embryonic development but bST did not affect the pregnancy rates of transferred embryos.

  5. 供体细胞来源和TSA处理对牦牛iSCNT胚胎重编程的影响%Effects of donor cell source and TSA treatment on reprogramming of yakbovine iSCNT embryos

    Institute of Scientific and Technical Information of China (English)

    熊显荣; 高川; 符梅; 李键; 字向东; 钟金城

    2012-01-01

    【Objective】The present study was to investigate the effect of different sources of donor cells and TSA treatment on in vitro development of yak interspecies somatic cell nuclear transfer(iSCNT)embryos.【Method】With cattle oocytes as recipient,yak fetal fibroblasts of 50-day-old,6 and 18-month-old adult fibroblasts as the donor nuclear,yak iSCNT embryos were reconstructed and then the effect of donor cell source on the development of cloned embryos evaluated.We compared the effect of different TSA treated concentrations(0,20,40,60 and 80 nmol/L)and times(0,6,8 and 12 h)on the development of yak cloned embryos,respectively,then determined the best program for treatment,and treated donor cells and embryos with this program for detecting the effect of TSA on the relative expression level of HDAC2 by qRT-PCR.【Result】With fetal fibroblast cells as donor cells,yak interspecies cloning embryos blastocyst rate was the highest,but there was no significant difference(P〉0.05).Treatment donor cell with 40 nmol/L TSA for 6 h significantly increased the blastocyst rate(30.6%),and significant difference existed with the control group(P〈0.05).Treatment of reconstructed embryos with 40 nmol/L TSA for 12 h was helpful for embryos development.However,the blastocyst formation rate decreased along with the treated time and concentration of TSA.The expression level of HDAC2 was reduced significantly after donor cell and embryos treated with TSA.【Conclusion】There was no significant effect of donor cells source on the development of yak iSCNT embryos.Treated donor cells for 6 h and reconstructed embryos for 12 h with 40 nmol/L TSA can significantly improve in vitro development of yak iSCNT embryos and reprogramming.%【目的】探讨不同来源的供体细胞和TSA处理对牦牛异种体细胞核移植(Interspecies nuclear transfer,iSCNT)胚胎的影响。【方法】以黄牛卵母细胞为受体,分别以牦牛50日龄胎儿及6和18月龄个体成纤维细

  6. IVF, same-sex couples and the value of biological ties

    DEFF Research Database (Denmark)

    Di Nucci, Ezio

    2016-01-01

    Ought parents, in general, to value being biologically tied to their children? Is it important, in particular, that both parents be biologically tied to their children? I will address these fundamental questions by looking at a fairly new practice within IVF treatments, so-called IVF......-with-ROPA (Reception of Oocytes from Partner), which allows lesbian couples to 'share motherhood', with one partner providing the eggs while the other becomes pregnant. I believe that IVF-with-ROPA is, just like other IVF treatments, morally permissible, but here I argue that the increased biological ties which IVF...

  7. A need for definition: a matter of life and death for human embryos

    OpenAIRE

    Shurpyak, Serhiy A; Walsh, Anthony PH; Walsh, David J.; Sills, E Scott

    2009-01-01

    A recent IMJ commentaryon brain stem death criteria summarised ethical and technical issues concerning “end of life decisions”, and we concur that physicians should have competence in eliciting the proper sequence of brain stem signs in clinical practice. However, a truly comprehensive dialogue on the definition of death should address another question that confronts IVF clinics in Ireland each day—when does a human embryo die? Despite the enormous social and political energy focused on “righ...

  8. Can Chlamydia abortus be transmitted by embryo transfer in goats?

    Science.gov (United States)

    Oseikria, M; Pellerin, J L; Rodolakis, A; Vorimore, F; Laroucau, K; Bruyas, J F; Roux, C; Michaud, S; Larrat, M; Fieni, F

    2016-10-01

    The objectives of this study were to determine (i) whether Chlamydia abortus would adhere to or penetrate the intact zona pellucida (ZP-intact) of early in vivo-derived caprine embryos, after in vitro infection; and (ii) the efficacy of the International Embryo Transfer Society (IETS) washing protocol for bovine embryos. Fifty-two ZP-intact embryos (8-16 cells), obtained from 14 donors were used in this experiment. The embryos were randomly divided into 12 batches. Nine batches (ZP-intact) of five embryos were incubated in a medium containing 4 × 10(7)Chlamydia/mL of AB7 strain. After incubation for 18 hours at 37 °C in an atmosphere of 5% CO2, the embryos were washed in batches in 10 successive baths of a phosphate buffer saline and 5% fetal calf serum solution in accordance with IETS guidelines. In parallel, three batches of ZP-intact embryos were used as controls by being subjected to similar procedures but without exposure to C. abortus. The 10 wash baths were collected separately and centrifuged for 1 hour at 13,000 × g. The washed embryos and the pellets of the 10 centrifuged wash baths were frozen at -20 °C before examination for evidence of C. abortus using polymerase chain reaction. C. abortus DNA was found in all of the infected batches of ZP-intact embryos (9/9) after 10 successive washes. It was also detected in the 10th wash fluid for seven batches of embryos, whereas for the two other batches, the last positive wash bath was the eighth and the ninth, respectively. In contrast, none of the embryos or their washing fluids in the control batches were DNA positive. These results report that C. abortus adheres to and/or penetrates the ZP of in vivo caprine embryos after in vitro infection, and that the standard washing protocol recommended by the IETS for bovine embryos, failed to remove it. The persistence of these bacteria after washing makes the embryo a potential means of transmission of the bacterium during embryo transfer from

  9. Oocytes with a dark zona pellucida demonstrate lower fertilization, implantation and clinical pregnancy rates in IVF/ICSI cycles.

    Directory of Open Access Journals (Sweden)

    Wei Shi

    Full Text Available The morphological assessment of oocytes is important for embryologists to identify and select MII oocytes in IVF/ICSI cycles. Dysmorphism of oocytes decreases viability and the developmental potential of oocytes as well as the clinical pregnancy rate. Several reports have suggested that oocytes with a dark zona pellucida (DZP correlate with the outcome of IVF treatment. However, the effect of DZP on oocyte quality, fertilization, implantation, and pregnancy outcome were not investigated in detail. In this study, a retrospective analysis was performed in 268 infertile patients with fallopian tube obstruction and/or male factor infertility. In 204 of these patients, all oocytes were surrounded by a normal zona pellucida (NZP, control group, whereas 46 patients were found to have part of their retrieved oocytes enclosed by NZP and the other by DZP (Group A. In addition, all oocytes enclosed by DZP were retrieved from 18 patients (Group B. No differences were detected between the control and group A. Compared to the control group, the rates of fertilization, good quality embryos, implantation and clinical pregnancy were significantly decreased in group B. Furthermore, mitochondria in oocytes with a DZP in both of the two study groups (A and B were severely damaged with several ultrastructural alterations, which were associated with an increased density of the zona pellucida and vacuolization. Briefly, oocytes with a DZP affected the clinical outcome in IVF/ICSI cycles and appeared to contain more ultrastructural alterations. Thus, DZP could be used as a potential selective marker for embryologists during daily laboratory work.

  10. THE PREDICTORS OF CLINICAL FERTILIZATION IN IVF CYCLES

    Directory of Open Access Journals (Sweden)

    S. Esmailzadeh

    2008-05-01

    Full Text Available We examined the demographic characteristics of couples, ovarian response and sperm quality to determine the predictors of clinical fertilization in IVF cycles. A descriptive and analytic study was conducted using data and biologic specimens obtained Fatemeh-Alzahra of Babol infertility center from 2004 to 2005. Only data from 315 women who had medical indication for conventional IVF treatment were included in the analysis. Treatment using ICSI was excluded. In the univariate analysis, the following variables affected on fertilization rate: the length of infertility, the number of IVF cycle, basal LH serum on 2 days, the number of administration of hMG, the duration of ovarian stimulation, the number of follicles, the number of oocytes retrieved, the number of oocytes stage II and III, sperm count, sperm motility, sperm grading III and IV. In the multivariate analysis, the strongest predictor of positive fertilization was the mean number of oocytes retrieved. Also, the mean number of oocytes stage II and stage III were positive predictors of fertilization. The mean of basal LH serum on day 2 and the mean duration of ovarian stimulation were negative predictors of fertilization Ovarian response to gonadotropins and the quality of oocytes were main predictors of fertilization. Although some parameters of sperm quality were significant variables of fertilization rate in univariate analysis, in multivariate analysis one's effects were negligible. This information should be used when selecting couples for IVF cycles or oocytes for fertilization to raise the rate of clinical fertilization.

  11. Psychosocial Adjustment to Unsuccessful IVF and GIFT Treatment.

    Science.gov (United States)

    Weaver, Susan M.; Clifford, Ellen; Hay, Douglas M.; Robinson, John

    1997-01-01

    Couples for whom in vitro fertilization (IVF) or gamete intrafallopian transfer (GIFT) treatment failed (N=20) were followed up and compared with successful couples. Current mental-health status, quality of life, and marital adjustment were assessed via questionnaires; experiences were explored by interview. Results and recommendations for…

  12. Melatonin in maturation media fails to improve oocyte maturation, embryo development rates and DNA damage of bovine embryos Melatonina no meio de maturação não melhorou as taxas de maturação dos ovócitos, de desenvolvimento embrionário e a fragmentação do DNA dos embriões bovinos

    Directory of Open Access Journals (Sweden)

    Luciana Takada

    2010-08-01

    Full Text Available Melatonin (MEL acts as a powerful scavenger of free radicals and direct gonadal responses to melatonin have been reported in the literature. Few studies, however, have evaluated the effect of MEL during in vitro maturation (IVM on bovine embryos. This study tested the addition of MEL to maturation medium (MM with no gonadotropins on nuclear maturation and embryo development rates and the incidence of DNA damage in resulting embryos. Cumulus-oocyte complexes were aspirated from abattoir ovaries and cultured in MM (TCM-199 medium supplemented with 10% fetal calf serum - FCS at 39ºC and 5% CO2 in air. After 24 hours of culture in MM with 0.5 µg mL-1 FSH and 5.0 µg mL-1 LH; 10-9 M MEL or 10-9 M MEL, 0.5 µg mL-1 FSH and 5.0 µg mL-1 LH, the oocytes were stained with Hoechst 33342 to evaluate nuclear maturation rate. After in vitro fertilization and embryo culture, development rates were evaluated and the blastocysts were assessed for DNA damage by Comet assay. There was no effect of melatonin added to the MM, alone or in combination with gonadotropins, on nuclear maturation, cleavage and blastocyst rates. These rates ranged between 88% to 90%, 85% to 88% and 42% to 46%, respectively. The extent of DNA damage in embryos was also not affected by MEL supplementation during IVM. The addition of 10-9 M MEL to the MM failed to improve nuclear maturation and embryo development rates and the incidence of DNA damage in resulting embryos, but was able to properly substitute for gonadotropins during IVM.Melatonin (MEL atua como um potente redutor de radicais livres. Efeito direto da MEL na função gonadal também foi observado. Existem poucos estudos relacionados ao efeito da MEL durante a maturação no desenvolvimento embrionário in vitro. Avaliou-se a adição de MEL no meio de maturação (sem gonadotrofinas nas taxas de maturação nuclear e de desenvolvimento embrionário e na incidência de fragmentação do DNA nos embriões produzidos in vitro

  13. Trichostatin A modified histone covalent pattern and enhanced expression of pluripotent genes in interspecies black-footed cat cloned embryos but did not improve in vitro and in vivo viability.

    Science.gov (United States)

    Gómez, Martha C; Pope, C Earle; Biancardi, Monica N; Dumas, Cherie; Galiguis, Jason; Morris, Anna Claire; Wang, Guoshun; Dresser, Betsy L

    2011-08-01

    Abstract The black-footed cat (BFC; Felis nigripes), one of the smallest wild cats, is listed as threatened. Interspecies somatic cell nuclear transfer (Is-SCNT) offers the possibility of preserving endangered species. Development to term of interspecies BFC (Is-BFC) cloned embryos has not been obtained, possibly due to abnormal epigenetic reprogramming. Treatment of intraspecies cloned embryos with TSA improves nuclear reprogramming and in vitro and in vivo viability. In this study, we evaluated (1) whether covalent histone modifications differ between Is-BFC cloned embryos and their IVF counterparts, (2) the optimal TSA concentration and exposure times to modify the covalent histone patterns, (3) if TSA enhances in vitro and in vivo developmental competence of cloned embryos, and (4) expression of pluripotent genes. Results indicated that the covalent histone modifications of Is-BFC cloned embryos aberrantly differ from their DSH-IVF counterpart embryos. Aberrant epigenetic events may be due partially to the inability of the DSH cytoplasm to modify the restrictive epigenetic marks of the BFC nuclei after somatic cell nuclear transfer (SCNT). Incomplete remodeling of the histone H3K9me2 in Is-BFC cloned embryos possibly contributes to abnormal expression of pluripotent genes and low embryonic development. Treatment of Is-BFC cloned embryos with TSA remodeled the covalent pattern in H3K9ac and H3K9me2, resembling epigenetic patterns in IVF counterpart embryos, and resulted in activation of some pluripotent genes. However, genomic reprogramming of Is-BFC cloned blastocysts did not follow the same reprogramming pattern observed in DSH-IVF embryos, and in vitro and in vivo developmental competence was not enhanced.

  14. Effect of Endometrial Injury on Secretion of Endometrial Cytokines and IVF Outcomes in Women with Unexplained Subfertility

    Directory of Open Access Journals (Sweden)

    Yu Liang

    2015-01-01

    Full Text Available In order to determine the effect of endometrial injury (EI on in vitro fertilization (IVF outcomes in women with unexplained subfertility and explore the relationship between EI and endometrial inflammatory cytokines, 66 women with unexplained subfertility undergoing IVF treatment were recruited. 38 patients in the EI group underwent EI in the mid-luteal phase of the cycle and 28 patients in the non-EI (NEI group. According to the pregnancy outcome, the NEI and EI groups were divided into NEI-nonpregnant (NEI-NP, NEI-pregnant (NEI-P, EI-NP, and EI-P. All patients underwent aspiration of endometrial secretions immediately before embryo transfer. The concentrations of ten mediators were measured using Milliplex Magnetic Bead assay. The clinical pregnancy was significantly higher in the EI than in the NEI group. The concentrations of interleukin- (IL- 6, IL-8, IL-12 (p70, IL-13, interferon- (IFN- γ, monocyte chemotactic protein- (MCP- 1, and vascular endothelial growth factor (VEGF were significantly higher in the EI than the NEI group. The expression of IFN-γ and VEGF in the EI-P was significantly increased compared to the EI-NP group. These findings suggest that, in women with unexplained subfertility, endometrial injury might be a potential method to improve clinical pregnancy rates by promoting the expression of IFN-γ and VEGF.

  15. Neuerungen im österreichischen IVF-Fonds- und Fortpflanzungsmedizingesetz sowie deren Folgerungen für die IVF-Institute

    Directory of Open Access Journals (Sweden)

    Tews G

    2005-01-01

    Full Text Available Seit dem 01.01.2000 regelt in Österreich das IVF-Fonds-Gesetz zusammen mit dem Fortpflanzungsmedizingesetz das Geschehen in der Reproduktionsmedizin. Im Vergleich mit umliegenden Staaten, insbesondere Deutschland, darf der Mediziner mit den Gegebenheiten durchaus zufrieden sein. Auf der einen Seite werden Medizin und Labortätigkeit in ihrem Wirken um das Wohl der Patienten nicht entscheidend eingeschränkt, auf der anderen Seite werden aber doch entscheidende ethisch-moralische Gesichtspunkte berücksichtigt. So ist es auf der einen Seite möglich, so viele Oozyten, wie für einen Zyklus sinnvoll ist, zu befruchten. Auf der anderen Seite sind Leihmutterschaften jedoch verboten. Mit 30.04.2004 wurde in Österreich das IVF-Fonds-Gesetz, das hauptsächlich die finanzielle Unterstützung der betroffenen Paare regelt, in mehreren entscheidenden Punkten abgeändert. Positiv zu vermerken ist, daß die wichtigen Sterilitätsfaktoren Endometriose und PCOS (Polycystisches Ovarsyndrom zukünftig ebenfalls einen Grund für die Unterstützung darstellen, auf der anderen Seite wurde der Zustand nach Tubenligatur und Vasektomie aus der Unterstützungsliste wiederum herausgenommen. Das abgeänderte Fortpflanzungsmedizingesetz (Änderung: 30.12.2004 regelt sowohl die Aufbewahrung von Ei- und Samenzellen, die nun bis auf Widerruf oder lebenslang aufbewahrt werden dürfen, als auch die Zeitdauer der Embryo-Konservierung, wobei nun dieses Zeitfenster von 1 auf 10 Jahre ausgedehnt worden ist. Gleichzeitig wurde auch die Anwendung der Fortpflanzungsmedizin bei Übertragungsgefahr von schweren Infektionskrankheiten legalisiert. Tatsächlich sind jedoch bei näherer Betrachtung Fragen offen geblieben bzw. Ungerechtigkeiten vorhanden. So ist nach Meinung des Autors die Definition einer erfolgreichen Schwangerschaft, die weitere 4 unterstützte Versuche nach sich zieht, nur unzureichend geklärt. Auch die Bedingung, eine Endometriose in jedem Fall durch eine

  16. Study of the Psychology of Pregnant Women who Prevented Miscarriage and Accepted Treatment with IVF-ET and the Nursing Countermeasures%体外受精-胚胎移植早孕保胎孕妇心理状态调查及护理对策

    Institute of Scientific and Technical Information of China (English)

    袁兰兰

    2013-01-01

    目的:分析体外受精-胚胎移植(IVF-ET)早孕保胎孕妇的心理状态,提出护理对策。方法采用心理状态90项症状自评量表(SCL -90)调查IVF-ET早孕保胎孕妇的心身症状水平,并对心身症状的频数进行分析。结果 IVF-ET早孕保胎孕妇的焦虑、抑郁、躯体化、敌对、强迫和人际关系敏感得分明显高于国内常模(P<0.05)。结论 IVF-ET早孕保胎孕妇存在明显的心理问题,需要采取相应的护理措施。%Objective To analyze the psychology of pregnant women who prevented miscarriage and accepted treatment with in vitro fertilization and embryo transfer (IVF-ET ) and raise the nursing countermeasures .Methods The symptom checklist 90(SCL -90)Scale about psychology was adopted to survey the psychology of pregnant women who prevented mis-carriage and accepted treatment with IVF -ET and the frequency of psychosomatic symptom was studied .Results The anx-iety ,depression ,somatization ,hostility ,obsession-compulsion and interpersonal sensitivity scores of the pregnant women who prevented miscarriage and accepted treatment with IVF -ET were remarkably higher than those in norm (P<0 .05) .Conclu-sion The pregnant women who prevented miscarriage and accepted treatment with IVF -ET showed obvious mental health problems ,and these mental health problems need to be dealt with the nursing countermeasures .

  17. The clinical analysis of the IVF-ET and FET pregnancy rate with thinner endometrial.%厚度偏薄的子宫内膜对体外受精-胚胎移植及冻融胚胎移植妊娠率的影响

    Institute of Scientific and Technical Information of China (English)

    张合龙; 李杭生; 张少娣; 郭海彬; 王雪; 何巧花; 路锦; 张翠莲

    2011-01-01

    探讨厚度偏薄的子宫内膜对体外受精-胚胎移植(in vitro fertilization and embryo transfer,IVF-ET)及冻融胚胎移植(freezing-thawing embryo transfer,FET)临床妊娠率的影响.方法2008年12月至2010年4月在河南省人民医院对218周期子宫内膜厚度在6 ~7.9mm之间的IVF-ET及FET患者进行回顾性分析,比较子宫内膜厚度在6~7.9mm之间的IVF-ET及FET患者的临床妊娠率有无差异.结果子宫内膜厚度在6~7.9mm之间的IVF-ET组与FET组患者的临床妊娠率分别为35.6%和50.7%,差异有统计学意义(P<0.05).结论 子宫内膜偏薄患者FET的临床妊娠率高于IVF-ET的临床妊娠率.%Objective To compare the pregnancy rate between IVF-ET patients and FET patients both with thinner endo-metrial thickness. Methods Between Dec 2008 and Apr 2010,We made retrospective clinical analysis to 218 ovulation induction cycles of IVF-ET and FET patients with endometrial thickness of 6 - 7. 9mm, in order to compare the pregnancy rate between these IVF-ET and FET patients in Henan Provincial Peoples Hospital. Results The pregnancy rate of IVF-ET group is 35. 6% while in FET group it is 50. 7% with significant differences (P < 0. 05). Conclusion Patients with thinner endometrial have higher pregnancy rates when FET than IVF-ET treated them.

  18. Superovulation of the Cloned Cattle Derived from Somatic Cells and the Transfer of the Vitrified-Thawed Embryos of the Cloning Cattle

    Institute of Scientific and Technical Information of China (English)

    DONG Ya-juan; BAI Xue-jin; LI Jian-dong; CHENG Ming

    2004-01-01

    In this experiment, it was designed to carry out superovulation on the two cloned cattles, vitrification and transfer of the embryos recovered from them. First of all, it was carried out vitrification on embryos obtained by IVF. Results showed that there were no significant differences between the blastocysts (obtained by IVF) vitrified in EPS10 and these in EPS20 on the resuscitative rate and the developmental rate. The hatched rate of the blastocysts vitrified in EPS10 (31.3%, 35/112) was significantly higher than that in EPS20 (12.2%, 13/107) (P<0.01), so EPS20 was selected as the vitrification solution to freeze the embryos recovered from the cloned cattle. After superovulation, six (four usable embryos) and ten (nine usable embryos) embryos were respectively recovered from Kangkang and Shuanghuang. Two embryos were selected from the recovered embryos of each cloned cattle to freeze in EPS20, subsequently thawed and transferred into luteal ipsilateral uterine horns of 4 Holstein recipient cows after synchronization of estrus, respectively. At last, one recipient cow (No. 9908) became pregnant and delivered one healthy calf (descendant of the cloned cattle-Shuangshuang). The results of this experiment show that the cloned cattle as well as common cattle had better response to the exotic FSH and better ability to multiovulation, the embryos recovered from the cloned cattle can be vitrificated.

  19. Effect of arachidonic acid supplementation and cyclooxygenase/lipoxygenase inhibition on the development of early bovine embryos Influência do ácido araquidónico e da inibição da ciclo-oxigenase ou lipo-oxigenase no desenvolvimento inicial de embriões bovinos

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    Rosa Maria Pereira

    2006-04-01

    Full Text Available The effect of arachidonic acid (AA cascade on bovine embryo development in a granulosa cell co-culture system was studied. Arachidonic acid (100 µM was supplemented from 1-cell to 8-16 cell block stage (first three days of co-culture and from 1-cell to hatching. Specific cyclooxygenase (indomethacin, 28 µM and lipoxygenase (nordihydroguaiaretic acid - NDGA, 28 µM inhibitors were used from 1-cell to 8-16 cell block stage with AA. Embryo development was assessed by cleavage, day 7-day 8 and hatched embryo rates and by measuring growth rates through development stages found in days 7-10 of culture (day 0 = insemination day. Embryo quality was scored at day 8. A 6.5-10.4% increase on cleavage rate after AA supplementation was found. This AA supplementation from 1-cell to hatching delayed embryo growth rate beyond day 7 and a reduction on hatching rate was detected. When AA supplementation was restricted to the first three days of co-culture those negative effects were overcome. Also, indomethacin and NDGA prevented the positive effect of AA and induced a significant reduction on cleavage, respectively. NDGA further decreased day 7 embryo rate and quality. Results suggest that AA has a two-phase action on bovine embryos, promoting early development and impairing embryo growth from day 7 onwards and hatching rates. Both cyclooxygenase and lipoxygenase were found to be important pathways to promote cleavage.Estudou-se a influência da cascata do ácido araquidónico (AA no desenvolvimento de embriões bovinos produzidos in vitro em co-cultura com células da granulosa. Os embriões foram suplementados com AA (100 µM desde o estádio de 1 célula até 8-16 células (primeiros três dias de co-cultura ou até a eclosão. Introduziram-se inibidores específicos da ciclo-oxigenase (indometacina, 28 µM e da lipo-oxigenase (ácido nordihidroguaiarético - NDGA, 28 µM, juntamente com o ácido araquidónico, desde o estádio de 1 célula até 8-16 c

  20. Does intrauterine saline infusion by intrauterine insemination (IUI) catheter as endometrial injury during IVF cycles improve pregnancy outcomes among patients with recurrent implantation failure?: An RCT

    Science.gov (United States)

    Salehpour, Saghar; Zamaniyan, Marzieh; Saharkhiz, Nasrin; Zadeh modares, Shahrzad; Hosieni, Sedighe; Seif, Samira; Malih, Narges; Rezapoor, Parinaz; Sohrabi, Mohammad-Reza

    2016-01-01

    Background: Recurrent implantation failure is one of the most issues in IVF cycles. Some researchers found that beneficial effects of endometrial Scratching in women with recurrent implantation failure, while some authors demonstrated contrary results Objective: The present study aimed to investigate the effect of intrauterine. Saline infusion as a form of endometrial injury, during fresh in vitro fertilization-embryo transfer cycle, among patients with recurrent implantation failure. Materials and Methods: In this clinical trial study 63 women undergoing assisted reproductive technology were divided into two groups either local endometrial injury by intrauterine saline infusion during day 3-5 of the ongoing controlled ovarian stimulation cycle, or IVF protocol performed without any other intervention in Taleghani Hospital, Tehran, Iran. The main outcome measure was clinical pregnancy rates. Results: Patients who received intra uterine saline infusion (n=20), had significantly lower clinical pregnancy numbers (1 vs. 9, p0.05) and multiple pregnancy numbers (1 vs. 3, p>0.05) between groups. Conclusion: When intrauterine saline infusion as a form of endometrial injury is performed during the ongoing IVF cycles it has negative effect on reproductive outcomes among patients with recurrent implantation failure. PMID:27738660

  1. A Prospective Randomized Controlled Study of Laser-Assisted Hatching on the Outcome of First Fresh IVF-ET Cycle in Advanced Age Women.

    Science.gov (United States)

    Shi, Wenhao; Hongwei, Tan; Zhang, Wei; Li, Na; Li, Mingzhao; Li, Wei; Shi, Juanzi

    2016-10-01

    There is no sufficient data to conclude the benefit of assisted hatching (AH) for advanced age patients. However, AH is routinely performed for advanced age patients undergoing in vitro fertilization (IVF) in China based on some retrospective evidence. It is important to assess the benefit of AH procedure for advanced age patients, especially by analyzing the data from China. This is a prospective randomized controlled trial to evaluate the effect of laser AH in the advanced age patients undergoing IVF. A total of 256 patients conformed to the inclusion criteria, and 78 were excluded by exclusion criteria. A total of 178 patients were eligible and randomized to 2 groups (82 AH group and 96 control group). Laser AH (zona thinning) was performed in the AH group. There were no statistical significance in basic clinical parameters between the 2 groups. No difference was found in implantation rate (AH vs control, 32.45% vs 39.29%) and clinical pregnancy rate (AH vs control, 48.78% vs 59.38%). Our data did not find any benefit of laser AH in improving implantation or pregnancy rates in advanced age women. Due to the potential risk and increasing financial burden, AH should not be routinely performed in first fresh IVF embryo transfer cycle for advanced age women.

  2. Global transcriptome profiles of Italian Mediterranean buffalo embryos with normal and retarded growth.

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    Maria Strazzullo

    Full Text Available The transcriptome profiles were compared for buffalo embryos with normal growth and embryos with retarded growth on Day 25 after mating. Embryos with retarded growth on Day 25 after mating have a reduced likelihood of undergoing attachment to the uterine endometrium and establishing a pregnancy. Italian Mediterranean buffaloes were mated by AI and on Day 25 underwent trans-rectal ultrasonography to ascertain embryo development. Embryos with an embryonic width (EW>2.7 mm were classed as normal embryos and embryos with an EW<2.7 mm were classed as retarded embryos. Three buffaloes with embryos of the largest EW (3.7, 3.7 and 3.9 mm and three buffaloes with embryos of the smallest EW (1.5, 1.6 and 1.9 mm were slaughtered on Day 27 to recover embryos for transcriptome analysis using a bovine custom designed oligo array. A total of 1,047 transcripts were differentially expressed between embryos with normal growth and embryos with retarded growth. Retarded embryos showed 773/1,047 (74% transcripts that were down-regulated and 274/1,047 (26% transcripts that were up-regulated relative to normal embryos; in silico analyses focused on 680/1,047 (65% of the differentially expressed transcripts. The most altered transcripts observed in retarded embryos were associated with membrane structure and function and with metabolic and homeostasis maintenance functions. Other notable functions altered in retarded embryos were developmental processes and in particular nervous system differentiation and function. Specific biochemical pathways such as the complement cascade and coagulation were also altered in retarded embryos. It was concluded from the findings that buffalo embryos with retarded growth on Day 25 after mating show altered gene expression compared with normal embryos, and some de-regulated functions are associated with attachment to the uterine endometrium.

  3. The Impact of Biopsy on Human Embryo Developmental Potential during Preimplantation Genetic Diagnosis

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    Danilo Cimadomo

    2016-01-01

    Full Text Available Preimplantation Genetic Diagnosis and Screening (PGD/PGS for monogenic diseases and/or numerical/structural chromosomal abnormalities is a tool for embryo testing aimed at identifying nonaffected and/or euploid embryos in a cohort produced during an IVF cycle. A critical aspect of this technology is the potential detrimental effect that the biopsy itself can have upon the embryo. Different embryo biopsy strategies have been proposed. Cleavage stage blastomere biopsy still represents the most commonly used method in Europe nowadays, although this approach has been shown to have a negative impact on embryo viability and implantation potential. Polar body biopsy has been proposed as an alternative to embryo biopsy especially for aneuploidy testing. However, to date no sufficiently powered study has clarified the impact of this procedure on embryo reproductive competence. Blastocyst stage biopsy represents nowadays the safest approach not to impact embryo implantation potential. For this reason, as well as for the evidences of a higher consistency of the molecular analysis when performed on trophectoderm cells, blastocyst biopsy implementation is gradually increasing worldwide. The aim of this review is to present the evidences published to date on the impact of the biopsy at different stages of preimplantation development upon human embryos reproductive potential.

  4. The Impact of Biopsy on Human Embryo Developmental Potential during Preimplantation Genetic Diagnosis

    Science.gov (United States)

    Cimadomo, Danilo; Capalbo, Antonio; Ubaldi, Filippo Maria; Scarica, Catello; Palagiano, Antonio; Canipari, Rita; Rienzi, Laura

    2016-01-01

    Preimplantation Genetic Diagnosis and Screening (PGD/PGS) for monogenic diseases and/or numerical/structural chromosomal abnormalities is a tool for embryo testing aimed at identifying nonaffected and/or euploid embryos in a cohort produced during an IVF cycle. A critical aspect of this technology is the potential detrimental effect that the biopsy itself can have upon the embryo. Different embryo biopsy strategies have been proposed. Cleavage stage blastomere biopsy still represents the most commonly used method in Europe nowadays, although this approach has been shown to have a negative impact on embryo viability and implantation potential. Polar body biopsy has been proposed as an alternative to embryo biopsy especially for aneuploidy testing. However, to date no sufficiently powered study has clarified the impact of this procedure on embryo reproductive competence. Blastocyst stage biopsy represents nowadays the safest approach not to impact embryo implantation potential. For this reason, as well as for the evidences of a higher consistency of the molecular analysis when performed on trophectoderm cells, blastocyst biopsy implementation is gradually increasing worldwide. The aim of this review is to present the evidences published to date on the impact of the biopsy at different stages of preimplantation development upon human embryos reproductive potential. PMID:26942198

  5. 如何进行IVF实验室的质控%Quality control in laboratory of IVF

    Institute of Scientific and Technical Information of China (English)

    刘东云; 韩伟; 黄国宁

    2010-01-01

    @@ 体外受精与胚胎移植(in vitro fertilization and embryo transfer,IVF-ET)是治疗不育症的有效手段,IVF实验室是其中极为重要的部分,对IVF结局有重要影响.在体内,配子和胚胎处于一个无光、恒温、恒湿、低氧的环境,母体内分泌及子宫/输卵管内分泌环境极其复杂,配子/胚胎在受到母体自身保护的环境下生长发育.

  6. Oocyte And Embryo Vitrification In The Ivf Laboratory: A Comprehensive Review

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    Simopoulou Maria

    2014-09-01

    Full Text Available Технология криоконсервации применяется с начала ХХ в. Открытие технологии витрификации повысило ожидания в области утвержденных клинических деятельностей в лабораториях по ин-витро фертилизации, привело к отличным результатам, как в отношении выживания после размораживания и количества беременностей, так и в отношении значительного улучшения клинических результатов преимплантационной генетической диагностики. В последнее время появились противоречивые утверждения относительно потенциала и ограничения применения витрификации в лабораторных условиях, а также относительно оптимального применения витрификации в области методик ин-витро фертилизации. Целью данной работы является предоставление подробного анализа практических сторон витрификации, в том числе и возможных рисков и вариантов её применения в отношении ооцитов и эмбрионов, по сравнению с традиционными методиками криоконсервации способом „медленного замораживания”. В заключение можно сказать, что для лабораторий ин-витро фертилизации витрификация является отличным средством, которое

  7. Metabolomics: approaches to assessing oocyte and embryo quality.

    Science.gov (United States)

    Singh, R; Sinclair, K D

    2007-09-01

    Morphological evaluation remains the primary method of embryo assessment during IVF cycles, but its modest predictive power and inherent inter- and intra-observer variability limits its value. Low-molecular weight metabolites represent the end products of cell regulatory processes and therefore reveal the response of biological systems to a variety of genetic, nutrient or environmental influences. It follows that the non-invasive quantification of oocyte and embryo metabolism, from the analyses of follicular fluid or culture media, may be a useful predictor of pregnancy outcome following embryo transfer, a potential supported by recent clinical studies working with specific classes of metabolites such as glycolytic intermediates and amino acids. Such selective approaches, however, whilst adhering closely to known cellular processes, may fail to harness the full potential of contemporary metabolomic methodologies, which can measure a wider spectrum of metabolites. However, an important technical drawback with many existing methodologies is the limited number of metabolites that can be determined by a single analytical platform. Vibrational spectroscopy methodologies such as Fourier transform infrared and near infrared spectroscopy may overcome these limitations by generating unique spectral signatures of functional groups and bonds, but their application in embryo quality assessment remains to be fully validated. Ultimately, a combination of evaluation criteria that include morphometry with metabolomics may provide the best predictive assessment of embryo viability.

  8. Molecular origin of mitotic aneuploidies in preimplantation embryos.

    Science.gov (United States)

    Mantikou, Eleni; Wong, Kai Mee; Repping, Sjoerd; Mastenbroek, Sebastiaan

    2012-12-01

    Mitotic errors are common in human preimplantation embryos. The occurrence of mitotic errors is highest during the first three cleavages after fertilization and as a result about three quarters of human preimplantation embryos show aneuploidies and are chromosomally mosaic at day three of development. The origin of these preimplantation mitotic aneuploidies and the molecular mechanisms involved are being discussed in this review. At later developmental stages the mitotic aneuploidy rate is lower. Mechanisms such as cell arrest, apoptosis, active correction of the aneuploidies and preferential allocation of the aneuploid cells to the extra-embryonic tissues could underlie this lower rate. Understanding the mechanisms that cause mitotic aneuploidies in human preimplantation embryos and the way human preimplantation embryos deal with these aneuploidies might lead to ways to limit the occurrence of aneuploidies, in order to ultimately increase the quality of embryos and with that the likelihood of a successful pregnancy in IVF/ICSI. This article is part of a Special Issue entitled: Molecular Genetics of Human Reproductive Failure. PMID:22771499

  9. Effect of Semen Quality on the Embryo Development

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    To investigate the influences of sperm quality on the zygotes and embryos development,as the role of the paternal factor in early human embryogenesis is gaining more attention because of the application of techniques such as intracytoplasmic sperm injection (ICSI) for the treatment of men infertility, 136 infertility couples with men factors (Group Ⅰ ) were included from May 2002 to January 2004. One hundred and seventy two infertility couples with tube factors (Group Ⅱ ) served as controls. The sperm parameters, gemmates and embryos quality, implantation rate and pregnant rate in both groups were analyzed. It was found that there was no significant differences in the number of oocytes retrieved, the fertilization rate and number of embryos transferred between two groups. Sperm concentration, percentage of motile sperm and percentage of sperm with normal morphology were significantly lower in group Ⅰ than in group Ⅱ (P<0.01). The proportion of good quality zygotes and good quality embryos were significantly lower in the male infertility group than in the tubal disease group (P<0.05). Implantation rate and pregnancy rate were similar in two groups. It was concluded that spermatozoa is involved in the embryo quality, even in the early stages of development, which limited the treatment potency of IVF procedure.

  10. Cryopreservation of animal oocytes and embryos: Current progress and future prospects.

    Science.gov (United States)

    Mandawala, A A; Harvey, S C; Roy, T K; Fowler, K E

    2016-10-15

    Cryopreservation describes techniques that permit freezing and subsequent warming of biological samples without loss of viability. The application of cryopreservation in assisted reproductive technology encompasses the freezing of gametes, embryos, and primordial germ cells. Whilst some protocols still rely on slow-freezing techniques, most now use vitrification, or ultra-rapid freezing, for both oocytes and embryos due to an associated decreased risk of damage caused by the lack of ice crystal formation, unlike in slow-freezing techniques. Vitrification has demonstrated its use in many applications, not only following IVF procedures in human embryology clinics but also following in vitro production of embryos in agriculturally important, or endangered animal species, before embryo transfer. Here, we review the various cryopreservation and vitrification technologies that are used in both humans and other animals and discuss the most recent innovations in vitrification with a particular emphasis on their applicability to animal embryology. PMID:27555525

  11. Applying embryo cryopreservation technologies to the production of domestic and black-footed cats.

    Science.gov (United States)

    Pope, C E; Gómez, M C; Galiguis, J; Dresser, B l

    2012-12-01

    Our objectives were (i) compare in vitro development of early cleavage stage domestic cat embryos after cryopreservation by minimal volume vitrification vs a standard slow, controlled-rate method, (ii) determine viability of vitrified domestic cat embryos by oviductal transfer into synchronous recipients and (iii) evaluate in vivo survival of black-footed cat (BFC, Felis nigripes) embryos after intra- and inter-species transfer. In vitro-derived (IVM/IVF) cat embryos were used to evaluate in vitro development after controlled-rate cryopreservation vs vitrification vs controls. Blastocyst development was similar in both groups of cryopreserved embryos (22-26%), but it was lower (p BFC, four intra-species embryo transfer procedures were carried out--two recipients received fresh day 2 embryos (n = 5, 8) and two recipients received embryos that had been cryopreserved on day 1 (n = 6) or 2 (n = 8). A 2-year-old recipient of cryopreserved embryos established pregnancy and delivered two live male kittens. Subsequently, five cryopreserved BFC embryos were transferred to a domestic cat recipient. On day 29, the recipient was determined to be pregnant and delivered naturally a live, healthy female BFC kitten on day 66. In summary, in vivo survival of vitrified domestic cat embryos was shown by the births of kittens after transfer into recipients. Also, we demonstrated that sperm and embryo cryopreservation could be combined with intra- and inter-species embryo transfer and integrated into the array of assisted reproductive techniques used successfully for propagation of a rare and vulnerable felid species, the black-footed cat.

  12. 长方案IVF/ICSI周期中获卵数与助孕结局的关系%Relationship between Numbers of Retrieved Oocytes and Outcomes of Assisted Reproduction in IVF/ICSI Cycles

    Institute of Scientific and Technical Information of China (English)

    倪佳; 黄静; 何钰辉; 柳胜贤

    2015-01-01

    目的:研究和分析长方案体外受精-胚胎移植(IVF/ICSI-ET)周期不同获卵数目与受精率、卵裂率、优质胚胎率、周期冷冻率、移植胚胎数、着床率、临床妊娠率之间的关系。方法:对本院生殖中心2011年1月-2012年12月实施长方案有获卵的共786个IVF/ICSI-ET新鲜周期进行回顾性总结和分析,根据不同获卵数分为五组(A组1~4枚,B组5~10枚,C组11~15枚,D组16~20枚,E组>20枚),比较各组年龄、受精率、卵裂率、优质胚胎率、周期冷冻率、移植胚胎数及临床妊娠率。结果:获卵数随年龄增加呈下降趋势,但五组年龄、移植胚胎数比较差异无统计学意义(P>0.05);五组受精率、卵裂率、优质胚胎率、冷冻周期率及临床妊娠率比较差异均有统计学意义(P0.05).However,the differences in fertilization rates, cleavage rates,good quality embryo rates,freezing cycle rates and clinical pregnancy rates of the five groups were statistically significant(P<0.01).Group C had the highest clinical pregnancy rates while group A had the lowest.The clinical pregnancy rates of group B and D were higher than those in group E.Conclusion:In IVF/ICSI cycles,if we have the same number of effectively transferred embryos,the number of retrieved oocytes is closely related to laboratory indexes and clinical pregnancy rates.The best clinical outcome can be reached by obtaining a modest number of oocytes.

  13. Economic optimization of the number of recipients in bovine embryo transfer programs Otimização econômica do número de receptoras em programas de transferência de embriões em bovinos

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    Renato Travassos Beltrame

    2007-06-01

    Full Text Available Purchase and maintenance of recipient females account for a large proportion of the costs and determine the number of calves that can be produced in an embryo transfer program. However, the large variability of embryo production by the donors and the need to purchase and synchronize the recipients before knowing the number of embryos collected make it difficult for the decision maker to identify the ideal number of recipient females to allocate. An ex-ante evaluation to determine the optimal number of recipient females was carried out through a sensitivity analysis for the ratio between the number of recipients and donors in a simulation model. The variability for the number of embryos collected was accounted for by applying the Monte Carlo simulation technique, assuming normal distribution and known values for mean and variance. The simulation considered monthly intervals between collections, during a 24 months program. The effect of embryo freezing on the number of pregnancies was considered by introducing a stock of frozen embryos into the mathematical model. Optimal recipient/donor ratio and the cost per pregnancy were compared for three recipient synchronization protocols (prostaglandin, progesterone - P4 and Ovsynch, based on the expected performance for synchronization, conception and transfer/treated rates for each protocol. Stochastic simulation associated with sensitivity analysis was effective in identifying the optimal donor to recipient ratio. Freezing embryos is effective to reduce the operational costs per pregnancy. The estimated optimal recipient/donor ratio was 20 for prostaglandin and 16.7 for the other protocols. The P4 protocol, although the most expensive, resulted in the lowest pregnancy cost estimation followed by prostaglandin and Ovsynch.A aquisição e manutenção de receptoras representam grande proporção dos custos e determinam o número de produtos gerados em um programa de transferência de embriões. Entretanto

  14. Establishment of Trophectoderm Cell Lines from Buffalo (Bubalus bubalis Embryos of Different Sources and Examination of In Vitro Developmental Competence, Quality, Epigenetic Status and Gene Expression in Cloned Embryos Derived from Them.

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    Sushil Kumar Mohapatra

    Full Text Available Despite being successfully used to produce live offspring in many species, somatic cell nuclear transfer (NT has had a limited applicability due to very low (>1% live birth rate because of a high incidence of pregnancy failure, which is mainly due to placental dysfunction. Since this may be due to abnormalities in the trophectoderm (TE cell lineage, TE cells can be a model to understand the placental growth disorders seen after NT. We isolated and characterized buffalo TE cells from blastocysts produced by in vitro fertilization (TE-IVF and Hand-made cloning (TE-HMC, and compared their growth characteristics and gene expression, and developed a feeder-free culture system for their long-term culture. The TE-IVF cells were then used as donor cells to produce HMC embryos following which their developmental competence, quality, epigenetic status and gene expression were compared with those of HMC embryos produced using fetal or adult fibroblasts as donor cells. We found that although TE-HMC and TE-IVF cells have a similar capability to grow in culture, significant differences exist in gene expression levels between them and between IVF and HMC embryos from which they are derived, which may have a role in the placental abnormalities associated with NT pregnancies. Although TE cells can be used as donor cells for producing HMC blastocysts, their developmental competence and quality is lower than that of blastocysts produced from fetal or adult fibroblasts. The epigenetic status and expression level of many important genes is different in HMC blastocysts produced using TE cells or fetal or adult fibroblasts or those produced by IVF.

  15. 多囊卵巢综合征行体外受精-胚胎移植助孕结局分析%The Outcome Analysis of Polycystic Ovary syndrome in Vitro Fertilization and Embryo Transfer Pregnancy

    Institute of Scientific and Technical Information of China (English)

    吴成平; 王芳

    2015-01-01

    目的:探讨多囊卵巢综合征(PCOS)伴不孕患者采用体外受精-胚胎移植(IVF-ET)助孕治疗的疗效及特点。方法:回顾分析2013年52周期PCOS不孕因素IVF-ET病历资料作为A组,同期102周期输卵管因素IVF-ET病例作对照作为B组。结果:PCOS组与输卵管因素组比较分析:患者年龄、不孕年限、BMI、Gn天数、获卵数、可移植胚胎数、OHSS风险取消移植率比较,差异有统计学意义(P0.05)。结论:PCOS伴不孕患者采用IVF-ET助孕可获得与输卵管因素IVF-ET助孕相似的疗效,但要注意OHSS的风险预防。%Objective:To evaluate the polycystic ovary syndrome (PCOS) with infertility were treated by in vitro fertilization-embryo transfer (IVF-ET) curative effect and characteristics of assisted reproduction treatment. Method: 52 cycles PCOS infertility factor in IVF-ET medical records in 2013 year were analysed retrospectively as group A, and 102 cycles tubal factor IVF-ET cases in the same period as controls of group B.Result:Compared PCOS group and tubal factor group analysis: age, infertility time, BMI, the number of eggs, portable designed the Gn days, embryo number and risk of OHSS cancel graft rate comparison, the difference was statistically significant(P<0.05). PCOS patients compared with tubal factor group more young, shorter infertility time, higher body mass index (BMI),longer Gn time,more egg number, more corresponding portable embryos and OHSS risk increased significantly. But two groups of designed the Gn dosage, endometrium thickness, 2 PN fertilization rate, cleavage rate, good embryo rate and the clinical pregnancy rate were no statistically significant difference.Conclusion:PCOS patients of infertility with the IVF-ET assist reproduction can be obtained IVF-ET assist similar curative effect with the tubal factor, but should pay attention to prevent the risk of OHSS.

  16. Successful pregnancy following the transfer of vitrified blastocyst which developed from poor quality embryos on day 3

    Directory of Open Access Journals (Sweden)

    Xiao-jian Zhang

    2011-01-01

    Full Text Available Background: The selection of pre-embryos for transferred is based on morphological appearance. But some poor quality cleaved embryos also can be cultured to the blastocyst stage and implanted.Objective: To assess the clinical pregnancy outcomes of blastocyst transfer which developed from poor quality embryos. Materials and Methods: A total of 109 cleaved embryos with poor quality were cultured to day 5/day 6 and 27 (24.8% blastocysts were collected from the 15 cycles/patients undergoing conventional IVF. All the blastocysts were cooling with fast-freezing. Then the blastocysts were warmed for transfer. Results: All of 25 vitrified blastocysts (92.6% survived after warming and were transferred to 15 patients. Five of the women became pregnant. Conclusion: Our results suggest that vitrified human day 5/day 6 blastocyst transfer which develop from poor quality embryo at day 3 can contribute to increasing cumulative pregnancy rates in assisted reproduction

  17. Chronological Reorganization of Microtubules, Actin Microfilaments, and Chromatin during the First Cell Cycle in Swamp Buffalo (Bubalus bubalis Embryos

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    Vibuntita Chankitisakul

    2010-01-01

    Full Text Available This paper aimed to study the dynamics of early embryonic development, in terms of redistribution of cytoskeleton (microtubules, actin microfilaments and chromatin configurations during the first cell cycle in swamp buffalo embryos. Oocytes were matured and fertilized in vitro, and they were fixed at various time points after IVF. At 6 h after IVF, 44.4% matured oocytes were penetrated by spermatozoa. Partial ZP digestion, however, did not improve fertilization rate compared to control (P>.05. At 12 h after IVF, the fertilized oocytes progressed to the second meiotic division and formed the female pronucleus simultaneously with the paternal chromatin continued to decondense. A sperm aster was observed radiating from the base of the decondensing sperm head. At 18 h after IVF, most presumptive zygotes had reached the pronuclear stage. The sperm aster was concurrently enlarged to assist the migration and apposition of pronuclei. Cell cleavage was facilitated by microfilaments and firstly observed by 30 h after IVF. In conclusion, the cytoskeleton actively involves with the process of fertilization and cleavage in swamp buffalo oocytes. The centrosomal material is paternally inherited. Fertilization failure is predominantly caused by poor sperm penetration. However, partial digestion of ZP did not improve fertilization rate.

  18. Mitochondrial functions on oocytes and preimplantation embryos

    Institute of Scientific and Technical Information of China (English)

    Li-ya WANG; Da-hui WANG; Xiang-yang ZOU; Chen-ming XU

    2009-01-01

    Oocyte quality has long been considered as a main limiting factor for in vitro fertilization (IVF). In the past decade,extensive observations demonstrated that the mitochondrion plays a vital role in the oocyte cytoplasm, for it can provide adenosine triphosphate (ATP) for fertilization and preimplantation embryo development and also act as stores of intracellular calcium and proapoptotic factors. During the oocyte maturation, mitochondria are characterized by distinct changes of their distribution pattern from being homogeneous to heterogeneous, which is correlated with the cumulus apoptosis. Oocyte quality decreases with the increasing maternal age. Recent studies have shown that low quality oocytes have some age-related dysfunctions, which include the decrease in mitochondrial membrane potential, increase of mitochondrial DNA (mtDNA) damages, chromosomal aneuploidies,the incidence of apoptosis, and changes in mitochondrial gene expression. All these dysfunctions may cause a high level of developmental retardation and arrest of preimplantation embryos. It has been suggested that these mitochondrial changes may arise from excessive reactive oxygen species (ROS) that is closely associated with the oxidative energy production or calcium overload,which may trigger permeability transition pore opening and subsequent apoptosis. Therefore, mitochondria can be seen as signs for oocyte quality evaluation, and it is possible that the oocyte quality can be improved by enhancing the physical function of mitochondria. Here we reviewed recent advances in mitochondrial functions on oocytes.

  19. Acute Transverse Myelitis Associated with Buserelin Use during IVF

    OpenAIRE

    Alleemudder, Djavid I.; Khaled Sadek; Shaun Fountain; Sallie Davies

    2013-01-01

    A healthy woman undergoing in vitro fertilization (IVF) developed acute transverse myelitis (ATM) following the use of Buserelin. ATM has a multifactorial etiology and may develop as a result of the activation of immune responses. Infectious agents have been postulated as possible triggers of an immune response (Sá, 2009). Gonadotropin-releasing agonists may have a similar role and trigger the acceleration of preexisting disease by the activation of immune responses (Ho et al., 1995, and Umes...

  20. Acute Transverse Myelitis Associated with Buserelin Use during IVF

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    Djavid I. Alleemudder

    2013-01-01

    Full Text Available A healthy woman undergoing in vitro fertilization (IVF developed acute transverse myelitis (ATM following the use of Buserelin. ATM has a multifactorial etiology and may develop as a result of the activation of immune responses. Infectious agents have been postulated as possible triggers of an immune response (Sá, 2009. Gonadotropin-releasing agonists may have a similar role and trigger the acceleration of preexisting disease by the activation of immune responses (Ho et al., 1995, and Umesaki et al., 1999.

  1. Besonderheiten der ektopen Schwangerschaft nach IVF/ICSI

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    Tews G

    2004-01-01

    Full Text Available Es ist allgemein bekannt, daß die weltweit erste Schwangerschaft nach In-Vitro-Fertilisation (IVF in einer Eileiterschwangerschaft endete [1]. Seit damals wurde in zahlreichen Arbeiten die Problematik der Extrauterinschwangerschaft im Rahmen der Reproduktionsmedizin beschrieben. Die Inzidenz schwankt je nach Arbeit zwischen 2 und 11 %. Teilweise konnte man keine Risikofaktoren entdecken, andere Autoren beschreiben erhöhte Inzidenzen bei kleineren Cavi uteri. Manchmal wurden beschädigte Tuben insbesondere für ektope Schwangerschaften als Risiko erwähnt. Das Ziel einer eigenen Untersuchung und einer Literaturanalyse war, die Inzidenz der ektopen Schwangerschaft, insbesondere der heterotopen Schwangerschaft (gleichzeitig intrauteriner und extrauteriner Sitz und deren Besonderheiten im Rahmen eines IVF/ICSI-Programms herauszuarbeiten. Die Untersuchung von 19 ektopen Schwangerschaften nach IVF aus dem eigenen Klientel und der Vergleich mit der Literatur hat ergeben, daß diese nach reproduktionsmedizinischen Eingriffen gehäuft vorkommen, wobei insbesondere tubare Faktoren mit nachfolgender IVF und weniger ICSI dafür verantwortlich sind. Von besonderer Wichtigkeit ist, daß zwar die Progesteronwerte im Vergleich zu spontan entstandenen Schwangerschaften erhöht sind, nicht jedoch β -hCG-Werte bzw. deren Anstieg. Weiters ist darauf zu achten, daß heterotope Schwangerschaften, also die Kombination von intra- und extrauterinen Graviditäten, ungleich häufiger vorkommen und in die diagnostischen Überlegungen unbedingt miteinbezogen werden müssen. Leider ist die Existenz einer derartigen heterotopen Schwangerschaft mit gleichzeitigem Vorkommen einer Überstimulation vor Eintritt einer Ruptur gelegentlich undiagnostizierbar.

  2. IVF-ET患者便秘的护理

    Institute of Scientific and Technical Information of China (English)

    胡冬玲; 李璟; 罗丽娟

    2009-01-01

    @@ IVF-ET患者中出现便秘症状比较常见,他不仅增加了患者心理上的痛苦,还对助孕治疗的效果产生不利影响,甚至导致严重的并发症,如引起脑出血、肠梗阻,甚至影响保胎效果而发生流产.

  3. In vivo versus in vitro produced bovine ova: similarities and differences relevant for practical application

    DEFF Research Database (Denmark)

    Holm, Peter; Callesen, Henrik

    1998-01-01

    - Abstract This present review describes some differences and similarities between bovine embryos produced in vivo and in vitro. The first part outlines the respective environments during maturation, fertilisation and early embryonic development of the two types of embryos and compares their mor-...

  4. Dexamethasone as a Supplement for Exogenous Gonadotropin to Improve Ovarian Response of Women over 35 Years Undergoing IVF/ICSI Cycles

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    Mahnaz Ashrafi

    2007-01-01

    Full Text Available Background: With aging, the ovarian reserve is decreased and that is a major contributor to poor ovarian response to exogenous gonadotropins. The aim of the present study is to evaluate the role of Dexamethasone on ovarian response in infertile patients aged over 35 years undergoing IVF/ICSI cycles.Materials and Methods: In this triple blind placbo-control clinical trial study, a total of 72 infertile women over age 35, undergoing IVF/ICSI cycles, referred to Royan Institute from May 2000 to May 2002 were selected. Dexamethasone co-treatment (1mg/d was started on the 21st of their preceding menstrual cycle and it was continued until oocyte aspiration. The main outcome measures were number of retrieved oocytes, number of fertilized and transferred embryos, number of used HMG, serum E2 level on HCG injection day, and pregnancy rate.Results: There was no significant statistical difference in age, duration of infertility, Body mass index, hormonal tests, number of retrieved oocytes and transferred embryos. However, the number of used HMG was significantly lower in Dexamethasone group compared to placebo group (30.6±13.39 versus 41.64 ± 18.34 (p<0.05.Conclusion: The addition of dexamethasone 1mg/d to standard long protocol decreased the number of HMG used in patients over 35 years who hold known risk of low ovarian response.

  5. Short communication. In vitro embryo production can be modified by the previous ovarian response to a superovulatory treatment in sheep

    Directory of Open Access Journals (Sweden)

    F. Forcada

    2013-04-01

    Full Text Available Thirty-two ewes were used to study how the ovarian response to a superovulatory treatment determines quality of oocytes recovered from ovaries after embryo collection, and their developmental capacity after in vitro maturation (IVM and fertilization (IVF. Ewes were superovulated, and seven days after oestrus, embryos were collected and ewes divided into three groups: (+ +, n=19, ewes responding to the treatment with embryos collected after flushing; (+ –, n=8, ewes responding, but only oocytes were found; and (– –, n=5, ewes not responding to the treatment and no embryos collected. Ovaries were recovered and oocytes collected from the three groups. A significant effect of the response to the treatment was observed for oocyte quality, so that (– – ewes presented the higher number of oocytes per ewe (p<0.001. Total number of oocytes selected for IVM and IVF was significantly higher in the same group, in comparison with (+ + and (+ – (p<0.001. Group (+ – ewes presented the lowest maturation (p<0.001, fertilization (p<0.05 and cleavage rates (p<0.001. In conclusion, the ovarian response to a superovulatory treatment determines the number and quality of the oocytes recovered 7 days after the oestrus induced by the hormonal treatment. In vitro techniques could be an important tool to increase embryo production by particular ewes when they are not able to produce a significant amount of in vivo embryos.

  6. Increasing The Number of Embryos Transferred from Two to Three, Does not Increase Pregnancy Rates in Good Prognosis Patients

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    Mahnaz Ashrafi

    2015-10-01

    Full Text Available Background: To compare the pregnancy outcomes after two embryos versus three embryos transfers (ETs in women undergoing in vitro fertilization (IVF/intracytoplasmic sperm injection (ICSI cycles. Materials and Methods: This retrospective study was performed on three hundred eighty seven women with primary infertility and with at least one fresh embryo in good quality in order to transfer at each IVF/ICSI cycle, from September 2006 to June 2010. Patients were categorized into two groups according to the number of ET as follows: ET2 and ET3 groups, indicating two and three embryos were respectively transferred. Pregnancy outcomes were compared between ET2 and ET3 groups. Chi square and student t tests were used for data analysis. Results: Clinical pregnancy and live birth rates were similar between two groups. The rates of multiple pregnancies were 27 and 45.2% in ET2 and ET3 groups, respectively. The rate of multiple pregnancies in young women was significantly increased when triple instead of double embryos were transferred. Logistic regression analysis indicated two significant prognostic variables for live birth that included number and quality of transferred embryos; it means that the chance of live birth following ICSI treatment increased 3.2-fold when the embryo with top quality (grade A was transferred, but the number of ET had an inverse relationship with live birth rate; it means that probability of live birth in women with transfer of two embryos was three times greater than those who had three ET. Conclusion: Due to the difficulty of implementation of the elective single-ET technique in some infertility centers in the world, we suggest transfer of double instead of triple embryos when at least one good quality embryo is available for transfer in women aged 39 years or younger. However, to reduce the rate of multiple pregnancies, it is recommended to consider the elective single ET strategy.

  7. Automated microinjection of recombinant BCL-X into mouse zygotes enhances embryo development.

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    Xinyu Liu

    Full Text Available Progression of fertilized mammalian oocytes through cleavage, blastocyst formation and implantation depends on successful implementation of the developmental program, which becomes established during oogenesis. The identification of ooplasmic factors, which are responsible for successful embryo development, is thus crucial in designing possible molecular therapies for infertility intervention. However, systematic evaluation of molecular targets has been hampered by the lack of techniques for efficient delivery of molecules into embryos. We have developed an automated robotic microinjection system for delivering cell impermeable compounds into preimplantation embryos with a high post-injection survival rate. In this paper, we report the performance of the system on microinjection of mouse embryos. Furthermore, using this system we provide the first evidence that recombinant BCL-XL (recBCL-XL protein is effective in preventing early embryo arrest imposed by suboptimal culture environment. We demonstrate that microinjection of recBCL-XL protein into early-stage embryos repairs mitochondrial bioenergetics, prevents reactive oxygen species (ROS accumulation, and enhances preimplantation embryo development. This approach may lead to a possible treatment option for patients with repeated in vitro fertilization (IVF failure due to poor embryo quality.

  8. Analysis of Factors Influencing Pregnancy Rate in Frozen-thawed Embryo Transfer

    Institute of Scientific and Technical Information of China (English)

    Lu LI; Xiao-xi SUN; Jun-ling CHEN; Xiao-hong GAO; Yong-wei WANG; Jie-wei TAO; Li-nan CHENG

    2004-01-01

    Objective To analyse factors influencing the outcome of frozen-thawed embryo transfer (FET)Method A retrospective analysis was performed in our center on 129 thawing cycles from March 2001 to April 2003. The related parameters were compared between conceived and non-conceived cycles.Results There were totally 129 clinical pregnancies in these transfers (pregnancy rate: 27.1%). Frozen-thawed embryos were transferred to natural cycles and CC cycling and hormone replacement treatment had equal success. Groups of IVF and ICSI did not differ significantly in pregnancy rates (P>0. 05). The pregnancy rates for one, two, three and four pre-embryos transfer were 0, 20.0%,44.1% and 75.0%,respectively (P<0. 05). There were statistical differences between pregnancy group or non- pregnancy group in the endometrial thickness, CES, CES/No. Of embryo. A higher pregnancy rate was observed in embryo transfers which had at least one 4-cell grade I embryo (d 2)(P<0.01). Conclusions The most important factors influencing the implantation rate and pregnancy rate of frozen-thawed embryo transfer are age, endometrium thickness, and the number, morphology and growth rate of transferred frozen embryos of women participants.

  9. Comparison of The Effectiveness of Clomiphene Citrate versus Letrozole in Mild IVF in Poor Prognosis Subfertile Women with Failed IVF Cycles

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    Mesut Oktem

    2015-10-01

    Full Text Available Background: Our objective was to evaluate the effectiveness of clomiphene citrate (CC vs. letrozole (L plus human menopausal gonadotropin (hMG in gonadotropin releasing hormone (GnRH antagonist protocol in poor prognosis women with previous failed ovarian stimulation undergoing intracytoplasmic sperm injection (ICSI. Materials and Methods: This retrospective cohort study included cycles with CC and L plus hMG/GnRH antagonist protocols of 32 poor responders who had failed to have ideal follicles to be retrieved during oocyte pick-up (OPU or embryo transfer (ET at least for 2 previous in vitro fertilization (IVF cycles with microdose flare protocol or GnRH antagonist protocol from January 2006 to December 2009. Main outcome measures were implantation, clinical pregnancy and live birth rates per cycle. Duration of stimulation, mean gonadotropin dose used, endometrial thickness, number of mature follicles, serum estradiol (E2 and progesterone (P levels on the day of human chorionic gonadotropin (hCG administration, number of retrieved oocytes and fertilization rates were also evaluated. Results: A total number of 42 cycles of 32 severe poor responders were evaluated. Total gonadotropin consumption was significantly lower (1491 ± 873 vs. 2808 ± 1581 IU, P=0.005 and mean E2 level on the day of hCG injection were significantly higher in CC group than L group (443.3 ± 255.2 vs. 255.4 ± 285.2 pg/mL, P=0.03. ET, overall pregnancy and live birth rates per cycle were significantly higher in CC than L protocol (27.2 vs. 15%, 13.6 vs. 0% and 4.5 vs. 0%, respectively, P<0.05. Conclusion: Severe poor responders who had previously failed to respond to microdose or GnRH antagonist protocols may benefit from CC plus hMG/GnRH antagonist protocol despite high cancellation rate.

  10. Fertilization and Embryo Development of Fresh and Cryopreserved Sibling Oocytes

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    Robert F. Casper

    2010-01-01

    Full Text Available Background: Oocyte cryopreservation is potentially the best way to preserve female fertility forunmarried women or young girls at risk of losing ovarian function. The aim of this study was tocompare fertilization and embryo development in frozen-thawed oocytes to their fresh siblings inwomen undergoing in vitro fertilization (IVF and embryo transfer (ET.Materials and Methods: Eleven infertile women undergoing infertility treatment, between theages of 24 to 37 years (mean ± SD = 31.6 ± 3.5, were included in this study. Mature oocytesfrom each patient were randomized into cryopreserved and fresh groups prior to intracytoplasmicsperm injection (ICSI. One hundred and thirty nine oocytes were retrieved, of which 105 were atmetaphase II (MII. Forty- five fresh MII oocytes were kept in culture whereas their sibling 60 MIIoocytes were cryopreserved using a slow cooling protocol. The frozen oocytes remained in LN2for 2 hours before thawing. ICSI was performed 1-2 hours after thawing for frozen oocytes and 4-5hours after retrieval for fresh oocytes. Fertilization and embryo development were compared.Results: Following thawing, 31 oocytes (51.6 % survived and 22 fertilized (79% while 32 freshoocytes fertilized upon ICSI (71%. The mean ± SE scores for embryos developing from frozenthawedoocytes were significantly lower at 48 and 72 hours post-ICSI than for embryos resultingfrom fresh oocytes (p<0.05.Conclusion: Our data demonstrated that oocyte freezing resulted in acceptable survival ratesfollowing cryopreservation, and similar fertilization rates following ICSI as compared to the freshsibling oocytes. However the number of blastomeres and the embryo quality on day three wassuperior in embryos from fresh oocytes when compared to the frozen oocytes.

  11. MODELO TEÓRICO PARA EXPLICAR LA ACUMULACIÓN DE GOTAS LIPÍDICAS EN EMBRIONES BOVINOS MACHOS O HEMBRAS PRODUCIDOS in vitro Theoretical Model For Explaining Accumulation Of Fat Drops In In Vitro Produced Bovine Embryos

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    OMAR CAMARGO

    Full Text Available La glucosa 6-fosfato deshidrogenasa (G6PD, codificada por un gen ubicado en el cromosoma X, es la enzima limitante de vía de las pentosas fosfato (PF. La entrada de la glucosa así como su flujo y el rendimiento metabólico de esta vía están determinados tanto por los mismos niveles glucosa así como por la actividad de la G6PD. Por esta vía, la glucosa regula la trascripción de varios genes lipogénicos. En algunos embriones hembra producidos in vitro, se registra un retardo en la normal inactivación de uno de sus cromosomas X, lo cual se traduce en una doble actividad de los genes allí ubicados, si se compara con los embriones macho producidos in vitro. Se postula entonces que, la sobre-regulación de la vía PF a consecuencia de la doble dosis de su enzima limitante (G6PD y en presencia de elevados niveles de glucosa (mayores a 2,5 mM en el medio de cultivo, conllevaría a un dimorfismo sexual en relación con la transcripción de los genes Acetil CoA Carboxilasa Alfa (en adelante ACACA, símbolo oficial de la acetyl-Coenzyme A carboxylase alpha, y la Sintetasa de Ácidos Grasos (en edelante FASN, símbolo oficial de la fatty acid synthase que corriente abajo codifican para las enzimas limitantes en la síntesis de lípidos. Este dimorfismo sexual para el fenotipo metabolismo de lípidos, derivaría en una mayor acumulación citoplasmática de gotas lipídicas en los embriones hembra en comparación con los embriones machos que, de ser así, tendría efectos expansivos sobre el metabolismo general, la actividad transcripcional de otros genes y sobre la resistencia a la criopreservación.The encoding gene for glucose 6-phosphate dehydrogenase (G6PD is located on chromosome X. This enzyme regulates the entrance of glucose into the pentose phosphate pathway (PPP. Besides, throughout this route, glucose regulates the transcription of some lipogenic genes. Compared with in vitro produced male embryos, and due to a delaying in X

  12. EFFECT OF BODY CONDITION AND SEASON ON THE YIELD AND QUALITY OF CATTLE EMBRYOS

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    Elena Kubovičová

    2013-02-01

    Full Text Available Unsatisfactory reproductive performance in dairy cows has been associated with environmental influences, such as season, chronic and acute changes in dietary intake and body composition. These factors can affect fertility especially ovarian function and yield and quality of oocytes and embryos. In our study the cow ´s body condition affected the overall embryo recovery rate (proportion of collected embryos to palpated corpora lutea. The significantly higher number of embryos was collected from cows with BCS 2.5- 2.75 (68.32 % embryo recovery rate and 3.0- 3.25 (63.30 % compared to the cows with BCS 2.0-2.25 (53.33% and 3.5-4.0 (47.87%; P0.05. On the contrary, the yield of transferable embryos was higher (P<0.05 during the autumn months (78.94% compared to spring (58.38% or summer (60.00% months. In conclusion, body condition and season may affect the yield and quality of bovine embryos. Higher embryo yield was recorded in average BCS (2.5-3.25 cows, whilst most transferable embryos were obtained in the higher BCS (3.5-4.0. Our results indicate that the best season for collection of transferable bovine embryos is autumn.

  13. Construction of Bovine (Bos taurus) Transgenic Cloned Embryos with Lysostaphin and Endolysin Genes by Electronic Transfection%电转染法制备奶牛转溶葡球菌酶(Lysostaphin)和内溶素(Endolysin)基因胚胎

    Institute of Scientific and Technical Information of China (English)

    杨林; 杜卫华; 郝海生; 刘岩; 秦彤; 赵学明; 王栋; 朱化彬; 王宗礼

    2013-01-01

    Lysostaphin is a single chain protease containing zinc which can kill staphylococcus aureus effectively. Endolysin which is the peculiar of the double-stranded DNA bacteriophages is a murein hydrolytic enzyme, it has a wide range of antibacterial effect. Lysostaphin and Endolysin have the high synergistic effect. In this study, the vectors pBCl-seq2 +seq3-EGFP-neo containing Endolysin and Lysostaphin genes and two other marker genes of enhenced green fluorescent protein (EGFP) andneomycin (neo) were transfected into bovine (Bos taurus) fetal fibroblast by electroporation and nucleofector of AMAXA. Stable transfected monoclonal cells which were identified to be the positive-cells in the way of PCR technique were obtained through fluorescence and G418 selection. Using transfected cells as the donor, transfected embryos were produced with somatic cell nuclear transfer, we used different conditions of AMAXA nuleofecor(A-023,V-013, V-023 and T-016) to transfect bovine fetal fibroblast, the results showed the suitable program was T-016. There were 5 times transfection efficient of AMAXA nuleofecor (20.11%) than it of electroporation. The blastocyst developed normally and the rate was of it 20.08%. In our study, we built up bovine fetal fibroblast cell line, sought out transfection parameter of high transfection efficiency, and acquired transgenic cell lines and transgenic blastocyst containing Lysostaphin and Endolysin genes, In conclusion, the results can provide technology supporting for producing anti-mastitis transgenic bovine and searching the new therapy way of mastitis.%溶葡球菌酶(Lysostaphin)是一种含锌的单链蛋白酶,能有效地杀灭金黄色葡萄球菌.内溶素(Endolysin)是双链DNA噬菌体所特有,是一类胞壁质水解酶,具有广泛的抗菌效果.内溶素与抗生素之间有高效的协同作用.本研究通过BTX电转染和AMAXA核转染的方法将含有溶葡球菌酶(Lysostaphin)和内溶素(Endolysin)两个目的基因(Seq2

  14. Oocyte activation and preimplantation development of bovine embryos obtained by specific inhibition of cyclin-dependent kinases Ativação oocitária e desenvolvimento pré-implantação de embriões bovinos obtidos com o uso de inibidores específicos das quinases dependentes de ciclina

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    F. Perecin

    2007-04-01

    Full Text Available The efficiency of bohemine and roscovitine in combination with ionomycin on parthenogenetic activation and initial embryonic development of bovine oocytes was studied. Two experiments were performed: in the first, different concentrations (0, 50, 75 or 100µM and different exposure periods (2, 4 or 6 hours to bohemine or roscovitine were tested for activation rates of in vitro matured (IVM bovine oocytes, which were pre-exposed to ionomycin. The best treatments, 75µM bohemine and 50µM roscovitine, both for 6h, were used in the second experiment, in which IVM bovine oocytes were exposed to ionomycin, followed or not by bohemine or roscovitine treatment, and evaluated for nuclear status, activation rate and blastocyst development were assessed. The combined treatments (ionomycin + cyclin-dependent kinases inhibitors - CDKIs showed better results for activation rates (77.3% and initial embryonic development (35.2% than the single ionomycin treatment (69.4% for activation and 21.9% for development; and also lead to a more uniform activation (nearly 90% single pronucleus development. The results showed that CDKIs improve the effects of ionomycin on parthenogenetic activation and blastocyst development in bovine oocytes and could help to achieve more efficient activation protocols, increasing the developmental competence of embryos obtained by reproductive biotechniques.Realizaram-se dois experimentos para avaliar a eficiência da bohemina e roscovitina associadas à ionomicina para ativação partenogenética e desenvolvimento embrionário inicial de bovinos. No primeiro, foram testadas diferentes concentrações (0, 50, 75 ou 100µM e diferentes tempos de exposição (2, 4 ou 6 horas à bohemina ou à roscovitina na ativação de oócitos bovinos maturados in vitro (MIV pré-expostos à ionomicina. Os melhores tratamentos, bohemina 75µM e roscovitina 50µM, ambos por seis horas, foram utilizados no segundo experimento, no qual oócitos bovinos

  15. Recent advances in in vitro embryo production in small ruminants.

    Science.gov (United States)

    Paramio, Maria-Teresa; Izquierdo, Dolors

    2016-07-01

    To increase productivity in the small ruminant industry, the genetic material of these species should be improved. In vitro embryo production could be an important technology to reach this goal by combining selected male and female gametes. In the world, marketing of in vitro-produced embryos is an economical activity which is progressing rapidly in cattle but is practically nonexistent in small ruminants. Since the birth of the first lamb and kid using IVF in the 80s, several studies have been carried out; however, results still are inconsistent and unpredictable. Moreover, significantly fewer research groups are working on embryo production in small ruminants than in cattle and pigs. Although conventional methodologies of oocyte IVM, IVF, and IVC in small ruminants give rise to blastocysts, significant variation exists between experiments. One important reason for these differences is the heterogeneity of the pool of oocytes recovered from ovaries from slaughtered females. Oocyte quality, also referred to as competence, is the key factor in the success of in vitro embryo production programs. Different criteria are used to select the best oocytes for fertilization, such as follicle size, oocyte diameter and morphological appearance, and Brilliant Cresyl Blue staining. New research lines aimed at improving oocyte competence are: (1) arresting nuclear maturation in vitro allowing optimal capacitation of cytoplasm, (2) growing oocytes inside the follicle, and (3) identification of biomarkers of oocyte competence in granulosa and cumulus cells and metabolites in the follicular fluid. PMID:27157391

  16. Application of hysteroscopy in 86 infertility patientients failed in IVF-ET%宫腔镜检查在IVF-ET失败患者中的应用

    Institute of Scientific and Technical Information of China (English)

    王宝金; 李根霞; 张金杏; 杨淑玲

    2010-01-01

    目的 探讨宫腔镜检查在IVF-ET失败患者中应用的重要性.方法 回顾分析2008年1月至2009年1月86例在IVF-ET失败后行宫腔镜检查的病例资料.结果 检查结果发现86例患者中57例有镜下异常,阳性率达66.28%.原发性不孕症宫腔镜检查异常为32例(61.54%),继发性不孕症宫腔镜检查异常为25例(73.53%),病变发生以子宫内膜息肉、子宫内膜炎及宫腔粘连为常见.结论 IVF-ET失败患者中宫腔病变发生率高,是导致IVF-ET失败的重要因素之一,在IVF-ET失败患者中有重要作用.%Objective To explore the application of hysteroscopy in women with inferlitily who failed in IVF-ET. Methods The clinical data of 86 cases of infertility patientients failed in IVF-ET in our Hospitall by using hysteroscopy were analysed between Jan and Dec. Results The incidence of uterine of 86 infertility patients was 66. 8%. The occurrence of uterine anormality in primary infrttility who failed IVF-ET was 61.54%,the secondary infertility patients was 73.53 %. The common disease was endometrial polpy, endometritis and uterine adhesion. Conclusion The rate of pathological changs of uterine cavity is high in infertility who failed in IVF-ET which is one of important infertile factors. Hysteroscopy has perfound useness in infertility clinic.

  17. Antiviral effects of bovine interferons on bovine respiratory tract viruses.

    OpenAIRE

    Fulton, R W; Downing, M M; Cummins, J M

    1984-01-01

    The antiviral effects of bovine interferons on the replication of bovine respiratory tract viruses were studied. Bovine turbinate monolayer cultures were treated with bovine interferons and challenged with several bovine herpesvirus 1 strains, bovine viral diarrhea virus, parainfluenza type 3 virus, goat respiratory syncytial virus, bovine respiratory syncytial virus, bovine adenovirus type 7, or vesicular stomatitis virus. Treatment with bovine interferons reduced viral yield for each of the...

  18. Discontinuation of rLH two days before hCG may increase the number of oocytes retrieved in IVF

    Directory of Open Access Journals (Sweden)

    Session Donna R

    2010-03-01

    Full Text Available Abstract Background Administration of recombinant luteinizing hormone (rLH in controlled ovarian hyperstimulation may benefit a subpopulation of patients. However, late follicular phase administration of high doses of rLH may also reduce the size of the follicular cohort and promote monofollicular development. Methods To determine if rLH in late follicular development had a negative impact on follicular growth and oocyte yield, IVF patients in our practice who received rFSH and rLH for the entire stimulation were retrospectively compared with those that had the rLH discontinued at least two days prior to hCG trigger. Results The two groups had similar baseline characteristics before stimulation with respect to age, FSH level and antral follicle count. However, the group which had the rLH discontinued at least two days prior to their hCG shot, had a significantly higher number of oocytes retrieved, including a higher number of MII oocytes and number of 2PN embryos. Conclusions When using rLH for controlled ovarian hyperstimulation, administering it from the start of stimulation and stopping it in the late follicular phase, at least two days prior to hCG trigger, may increase oocyte and embryo yield.

  19. Early embryo development in a sequential versus single medium: a randomized study

    Directory of Open Access Journals (Sweden)

    D'Hooghe Thomas M

    2010-07-01

    Full Text Available Abstract Background The success of in vitro fertilization techniques is defined by multiple factors including embryo culture conditions, related to the composition of the culture medium. In view of the lack of solid scientific data and in view of the current general belief that sequential media are superior to single media, the aim of this randomized study was to compare the embryo quality in two types of culture media. Methods In this study, the embryo quality on day 3 was measured as primary outcome. In total, 147 patients younger than 36 years treated with IVF/ICSI during the first or second cycle were included in this study. Embryos were randomly cultured in a sequential (group A or a single medium (group B to compare the embryo quality on day 1, day 2 and day 3. The embryo quality was compared in both groups using a Chi-square test with a significance level of 0.05. Results At day 1, the percentage of embryos with a cytoplasmic halo was higher in group B (46% than in group A (32%. At day 2, number of blastomeres, degree of fragmentation and the percentage of unequally sized blastomeres were higher in group B than in group A. At day 3, a higher percentage of embryos had a higher number of blastomeres and unequally sized blastomeres in group B. The number of good quality embryos (GQE was comparable in both groups. The embryo utilization rate was higher in group B (56% compared to group A (49%. Conclusions Although, no significant difference in the number of GQE was found in both media, the utilization rate was significantly higher when the embryos were cultured in the single medium compared to the sequential medium. The results of this study have a possible positive effect on the cumulative cryo-augmented pregnancy rate. Trial registration number NCT01094314

  20. Clinical observation on preventing ovarian hyperstimulation syndrome by frozen-thawed embryo transfer after all embryo freezing%全胚冷冻后行冻融胚胎移植预防卵巢过度刺激综合征的临床观察

    Institute of Scientific and Technical Information of China (English)

    黄永刚; 黄朝霞; 侯晓红; 肖宇; 吕杰强

    2012-01-01

    Objective: To compare the clinical pregnancy outcomes of frozen - thawed embryo transfer (FET) after all embryo freezing and routine in -vitro fertilization and embryo transfer (IVF - ET) , and explore the clinical application for prevention of ovarian hyperstimulation syndrome ( OHSS) . Methods: The clinical data of 124 patients receiving FET to prevent moderate and severe OHSS and 987 patients receiving routine long protocol of IVF — ET were analyzed retrospectively, ihe embryo implantation rates and clinical pregnancy rates in the two groups were compared. Results: There was no statistically significant difference in age, duration of infertility, body mass index, the number of transferred embryos, embryo implantation rate, and clinical pregnancy rate between the two groups ( P > 0. 05 ) , After IVF -ET, 25 patients in patients receiving IVF, went to hospital for treatment because of severe OHSS, and no one in patients receiving FET went to hospital. Conclusion: FET after all embryo freezing can achieve a good pregnancy outcome, which is a safe clinical application scheme for preventing moderate and severe OHSS.%目的:比较全胚冷冻后行冻融胚胎移植(FET)和常规体外受精-胚胎移植(IVF-ET)的临床妊娠结局,探讨预防卵巢过度刺激综合征(OHSS)的临床应用.方法:回顾性分析1VF-ET过程中为防止中、重度OHSS而行FET的124例患者和常规长方案IVF-ET987例患者的临床资料,比较其胚胎种植率、临床妊娠率.结果:2组的年龄、不孕年限、体重指数差异无统计学意义(P>0.05),移植胚胎个数、胚胎种植率、临床妊娠率差异亦无统计学意义(P>0.05).IVF组移植后有25例患者因重度OHSS发生而住院治疗,冷冻组无一例住院治疗.结论:全胚冷冻后行冻融胚胎移植可以获得良好的妊娠结局,是一项安全的预防中、重度卵巢过度刺激综合征的临床应用方案.

  1. 人精子-透明质结合试验与IVF-ET结局相关性研究%Relationship between human sperm-hyaluronan binding assay and IVF-ET outcomes

    Institute of Scientific and Technical Information of China (English)

    王金香; 邱孙全; 叶英辉; 金帆

    2015-01-01

    目的:分析精子-透明质结合试验(hyaluronan binding assay,HBA)与精子常规参数及体外受精-胚胎移植(in vitro fertilization-embryo transfer,IVF-ET)结局的相关性,探寻精子质量评估和授精方式选择的新指标.方法:对进入IVF-ET周期的136例患者进行HBA检测,并按HBA结合率将患者分为HBA结合率≥65%组(A组)与HBA结合率<65%组(B组),统计分析HBA与精液常规参数、体外受精率、胚胎质量和临床妊娠结局的相关性.结果:A组与B组相比,精子浓度无统计学差异(P>0.05);而前者的前向运动(PR)精子明显高于后者,差异有显著统计学意义(P<0.01).A组总受精率和正常受精率均高于B组(P<0.01),而可利用胚胎率、临床妊娠率、胚胎着床率及活产婴儿出生率组间均无统计学差异(P>0.05).结论:HBA反映精子活动力,对体外受精率具有指示作用,对于临床选择体外辅助授精方式具有一定的参考意义.

  2. The relative contribution of IVF to the total ongoing pregnancy rate in a subfertile cohort.

    NARCIS (Netherlands)

    Brandes, M.; Hamilton, C.J.C.M.; Bruin, J.P. de; Nelen, W.L.D.M.; Kremer, J.A.M.

    2010-01-01

    BACKGROUND: Although in vitro fertilization (IVF) was introduced more than 30 years ago, its exact role in the spectrum of fertility treatments has never been studied in an unselected population. The aim of this study was to visualize the contribution of IVF to the ongoing pregnancy rates in a cohor

  3. New Reproductive Assemblages: Understanding, Managing and ‘Using’ Human In Vitro Fertilization (IVF)

    NARCIS (Netherlands)

    Just, E.M.

    2009-01-01

    This dissertation is a contribution to the ongoing discussion about the body and in vitro fertilization (IVF), also known as assisted reproduction or technologically enhanced reproduction. With help of empirical research on Dutch and Polish IVF-couples, Edyta Just puts into question the meaning of I

  4. Effectiveness of a psychosocial counselling intervention for first-time IVF couples : a randomized controlled trial

    NARCIS (Netherlands)

    de Klerk, C; Hunfeld, JAM; Duivenvoorden, HJ; den Outer, MA; Fauser, BCJM; Passchier, J; Macklon, NS

    2005-01-01

    BACKGROUND: The objective of this study was to evaluate a psychosocial counselling intervention for first-time IVF couples. In this article the results on women's distress are presented. METHODS: Two hundred sixty-five couples admitted to an IVF treatment programme at the Erasmus MC were asked to pa

  5. Vanishing twins: a predictor of small-for-gestational age in IVF singletons

    DEFF Research Database (Denmark)

    Pinborg, Anja; Lidegaard, Ojvind; Freiesleben, Nina la Cour;

    2007-01-01

    The purpose of this study was to assess the effect of a vanishing twin on the risk of being small-for-gestational age (SGA) in in vitro fertilization (IVF) singletons.......The purpose of this study was to assess the effect of a vanishing twin on the risk of being small-for-gestational age (SGA) in in vitro fertilization (IVF) singletons....

  6. Comparative evaluation of growth parameters of IVF children of West Bengal

    Directory of Open Access Journals (Sweden)

    Sudipta Kar

    2014-01-01

    Full Text Available Context: Fertility rates have started declining in West Bengal, India in the last few decades. In vitro fertilization (IVF is one of the major treatment modalities available to infertile couple. Aims: This study was aimed to evaluate and compare the growth parameters of IVF and spontaneously conceived children. The objective of this study is to relate the physical status of IVF children with that of spontaneously conceived group. Settings and Design: In a cross-sectional case-control study, growth parameters of 3-14-year-old children were assessed. Subjects and Methods: Weight and height of 3-14-year-old children were assessed. Birth weight and length of each case and control group was also recorded. The case group consisted of term, singleton babies who were outcome of IVF in the studied area in 2009. The control group consisted of term, first child, singleton, and spontaneously conceived 3-14-year-old children who were also resident of the studied area. A sample of 150 IVF and 150 spontaneously conceived children were examined. Statistical Analysis Used: Chi-square and student′s t-test were used for data analysis. Results: No statistically significant difference found in the studied (IVF children and the control group (spontaneously conceived children. Conclusions: Growth retardation is more prevalent in babies born through IVF in the initial phase of life, but later on, IVF children are considered same as spontaneously conceived children.

  7. Value of ovarian reserve testing before IVF: a clinical decision analysis

    NARCIS (Netherlands)

    B.W. Mol; T.E.M. Verhagen; D.J. Hendriks; J.A. Collins; A. Coomarasamy; B.C. Opmeer; F.J. Broekmans

    2006-01-01

    BACKGROUND: To assess the value of testing for ovarian reserve prior to a first cycle IVF incorporating patient and doctor valuation of mismatches between test results and treatment outcome. METHODS: A decision model was developed for couples who were considering participation in an IVF programme. T

  8. Cryorecovery of Mouse Sperm by Different IVF Methods Using MBCD and GSH

    Science.gov (United States)

    Li, Ming-Wen; Glass, Olivia C; Zarrabi, Jasmin; Baker, Lisa N.; Lloyd, K. C. Kent

    2016-01-01

    Different protocols incorporating methyl-β-cyclodextrin (MBCD) and reduced glutathione (GSH) have been reported to improve IVF recovery of cryopreserved mouse sperm on a C57BL/6 (J and N) genetic background. However, it is not clear which IVF protocol is most appropriate when using the various methods to cryorecover sperm with different sperm quality and sample volumes. Therefore, in the present study we correlated sperm motility with fertilization rate and compared the efficiency of different IVF methods using various sperm samples so as to establish general guidelines for mouse sperm cryorecovery by IVF. High linear correlation between sperm fertilization rate and progressive motility was found, R2 was 0.9623 and 0.9993 for pre-freezing and post-thaw progressive motility, respectively. High amounts of cryoprotective agent (CPA) were observed to impair both sperm capacitation and fertilization. Moreover, the presence of a large number of immotile sperm in the sperm-oocyte co-incubation drop was found to reduce IVF success which could be partially reversed by supplementation using monothioglycerol (MTG) during centrifugation. It was concluded that the efficiency of IVF using cryorecovered mouse sperm in media containing MBCD and GSH can be predicted from sperm progressive motility. High concentrations of CPA and immotile sperm should be mitigated prior to IVF. The optimum IVF method should be selected based on sperm sample volume and sperm parameters. PMID:27413624

  9. Effects of subfertility cause, smoking and body weight on the success rate of IVF

    NARCIS (Netherlands)

    Lintsen, A.M.E.; Pasker-Jong, P.C.M. de; Boer, E.J. de; Burger, C.W.; Jansen, C.A.M.; Braat, D.D.M.; Leeuwen, F.E. van; Kortman, M.; Velde, E.R. te; Macklon, N.; Jansen, C.A.M.; Leerentveld, R.A.; Willemsen, W.N.P.; Schats, R.; Naaktgeboren, N.; Helmerhorst, F.M.; Bots, R.S.G.M.; Simons, A.H.M.; Hogerzeil, H.V.; Evers, J.L.H.; Dop, P.A. van

    2005-01-01

    Background: We investigated the separate and combined effects of smoking and body mass index (BMI) on the success rate of IVF for couples with different causes of subfertility. Methods: The success rate of IVF was examined in 8457 women. Detailed information on reproduction and lifestyle factors was

  10. Effects of subfertility cause, smoking and body weight on the success rate of IVF.

    NARCIS (Netherlands)

    Lintsen, A.M.E.; Pasker-de Jong, P.C.M.; Boer, E.J. de; Burger, C.W.; Jansen, C.A.M.; Braat, D.D.M.; Leeuwen, F.E. van

    2005-01-01

    BACKGROUND: We investigated the separate and combined effects of smoking and body mass index (BMI) on the success rate of IVF for couples with different causes of subfertility. METHODS: The success rate of IVF was examined in 8457 women. Detailed information on reproduction and lifestyle factors was

  11. Incidence of retinoblastoma in Dutch children conceived by IVF: an expanded study.

    NARCIS (Netherlands)

    Marees, T.; Dommering, C.J.; Imhof, S.M.; Kors, W.A.; Ringens, P.J.; Leeuwen, F.E. van; Moll, A.C.

    2009-01-01

    BACKGROUND: In 2003, we reported an increased risk of retinoblastoma in children conceived by IVF between 1995 and 2002. However, population-based studies among children conceived by IVF did not find an elevated risk of retinoblastoma. METHODS: From nationwide estimates of numbers of live births con

  12. Classification of Patients Treated for Infertility Using the IVF Method

    Directory of Open Access Journals (Sweden)

    Malinowski Paweł

    2015-12-01

    Full Text Available One of the most effective methods of infertility treatment is in vitro fertilization (IVF. Effectiveness of the treatment, as well as classification of the data obtained from it, is still an ongoing issue. Classifiers obtained so far are powerful, but even the best ones do not exhibit equal quality concerning possible treatment outcome predictions. Usually, lack of pregnancy is predicted far too often. This creates a constant need for further exploration of this issue. Careful use of different classification methods can, however, help to achieve that goal.

  13. Behandlungskostenerstattung bei IVF/ICSI in Deutschland - eine aktuelle Betrachtung

    Directory of Open Access Journals (Sweden)

    Nauert R

    2007-01-01

    Full Text Available Die aktuelle Behandlungskostenerstattung bei IVF/ICSI in Deutschland wird nach wie vor durch die unterschiedlichen Versicherungsprinzipien der Körperbezogenheit in der GKV und der Verursacherbezogenheit in der PKV und damit einhergehenden Lücken der Kostenerstattung geprägt. Durch die Änderung des § 27a SGB V wurden die Leistungen der GKV vom Gesetzgeber zudem erheblich eingeschränkt, die Leistungen der PKV andererseits sind von der Rechtsprechung erheblich ausgeweitet worden, was zu einer Verschärfung der Problematik geführt hat.

  14. Resumption of mitosis in frozen-thawed embryos is not related to the chromosomal constitution

    DEFF Research Database (Denmark)

    Agerholm, Inge E; Kølvrå, Steen; Crüger, Dorthe G;

    2007-01-01

    OBJECTIVE: To study the relation between the resumption of mitosis after thaw and chromosomal constitution in frozen-thawed embryos. In addition, to evaluate the correlation among the three parameters of resumption of mitosis after thaw, postthaw blastomere loss, and multinucleation. DESIGN: Frozen......-thawed embryos were morphologically evaluated at thaw and after 24 hours of culture. Then, fluorescence in situ hybridization (FISH) analysis, including enumeration of 13 chromosomes, was performed by using a combination of peptide nucleic acid and DNA probes. SETTING: In vitro fertilization laboratory. PATIENT......(S): Forty IVF and/or intracytoplasmic sperm injection patients. INTERVENTION(S): Embryo thawing, morphological evaluation, and fluorescence in situ hybridization analysis for aneuploidy screening. MAIN OUTCOME MEASURE(S): Resumption of mitosis, blastomere loss, multinucleation, and chromosome enumeration...

  15. 口服避孕药在IVF-ET短方案促排卵中的作用%The role of oral contraceptives in the GnRH-a flare-up protocol in IVF-ET

    Institute of Scientific and Technical Information of China (English)

    钱秋然; 孙丽君; 王兴玲; 孙慧霞

    2011-01-01

    目的 观察口服避孕药OCP)在体外受精-胚胎移植(IVF-ET)短方案促排卵中的作用.方法 对因输卵管因素或男方因素行IVF-ET的不孕患者共112例,均采用短方案促排卵,其中实验组59例,于促排卵前给予OCP口服1个月;对照组53例,促排卵前不给予OCP预处理,常规月经第3天促排卵.比较两组患者自然月经周期D3天和COH周期月经D3天的基础卵泡及内分泌情况;比较两组Gn使用天数、Cn总量、HCG日LH、E2、P水平;比较两组取卵日成熟卵泡情况、获卵数、受精率、卵裂率、可用胚胎数、优质胚胎数、周期取消率、临床妊娠率的差异.结果 ①两组患者的年龄、不孕年限、自然月经周期基础FSH、LH、E比较无显著性差异(均P>0.05);两组患者COH周期月经D3天FSH、LH、E2水平及HCG日LH、E2、P水平比较无显著性差异(均P>0.05);②COH周期月经D3天有>5mm卵泡的患者数,实验组较对照组明显减少(p0.05).结论 在IVF-ET短方案中,在GnRH-a+Gn促排卵前用OCP预处理,能调整月经周期,使卵泡发育同步性增加,能获得比较好的促排卵效果,取卵日成熟卵泡数及获得的卵子数均较多,周期取消率降低,临床妊娠率能够得到适当的提高.但OCP预处理后可能抑制了早期内源性FSH的上升,对卵巢的抑制加重,使小卵泡发育缓慢而用药时间延长.%Objective: To observe the role of oral contraceptives (OCP) in the GnRH -a flare -up protocol in in -vitro fertilization - embryo transfer ( IVF - ET).Methods: The objects of the study were 112 cases with infertility for tubal factor or male factor and they were dealed with IVF - ET.The protocol of COH was GnRH - a short flare - up protocol.All of the 112 cases were divided into two groupes: the experimental group with 59 cases who were given oral contraceptives one month before COH; the controlled group with 53 cases who were not given oral contraceptives one month before COH.Compare follicular

  16. 心理干预对IVF-ET疗效的促进作用%The Pregnancy Improvement of IVF-ET by Mental Intervention

    Institute of Scientific and Technical Information of China (English)

    唐向辉; 陈晓刚; 张琼芬; 叶少玲; 曹跃龄

    2013-01-01

    目的:探讨在IVF-ET治疗过程中,应用心理干预措施提高IVF-ET成功率.方法选取2012年2月至2013年1月间在云南省第二人民医院接受IVF-ET技术治疗的不孕症患者206例,随机分为研究组与对照组,各103例,研究组在进行IVF-ET治疗前后给予心理干预措施,并用汉密顿焦虑量表(HAMA)和汉密顿抑郁量表(HAMD)对患者进行心理状态评分.结果研究组心理干预后的焦虑抑郁评分比心理干预前较低,前后比较,差异有统计学意义(<0.01),研究组的焦虑抑郁评分比对照组的显著低(<0.01),研究组的妊娠率(45.63%)与对照组(30.10%)比较,差异有统计学意义(<0.05).结论心理干预对IVF-ET疗效有明显的促进作用.%Objective To study the effect of mental intervention on pregnant result in patients undergoing IVF-ET treatment.Methods 206 patients undergoing IVF-ET treatment in the hospital from February 2012 to Jan-uary 2013 were selected and randomly divided into two groups. The experiment group (n=103) was followed by mental intervention after the patients enrolled the IVF-ET treatment, the control group (n=103) enrolled the IVF-ET treatment without mental intervention.Results The HAMA and HAMD quantity grades for anxious and depression of patients in the experiment group were significantly lower than those of control ( <0.01), and the pregnancy rate of experiment group (45.63%) was higher than that of control (30.10%) <0.05. Conclusion Mental intervention for patients undergoing IVF-ET treatment could alleviate their anxiety-depression and improve the pregnancy rate of IVF.

  17. Serum dehydroepiandrosterone sulphate concentration is not a predictive factor in IVF outcomes before the first cycle of GnRH agonist administration in women with normal ovarian reserve.

    Directory of Open Access Journals (Sweden)

    Michał Kunicki

    Full Text Available The aim of our study was to determine whether serum dehydroepiandrosterone sulphate (DHEAS concentration and the models incorporating it could help clinicians to predict IVF outcomes in women with normal ovarian reserve undergoing their first long protocol.We performed a retrospective analysis of 459 women undergoing cycles of intracytoplasmic sperm injection (ICSI for the first time in a long GnRH agonist protocol.Embryo transfer was performed in 407 women (88.7%. The fertilisation rate was 78.6%. The clinical pregnancy rate was 44.8% per started cycle and 50.6% per embryo transfer. Our univariate model revealed that the best predictors of clinical pregnancy were the number of mature oocytes, the number of embryos transferred and the number of good quality embryos, account for the clinical parameters that reflect ovarian reserve the best being AMH level and AFC. DHEAS did not predict clinical pregnancy (OR 1.001, 95% CI, 0.999-1.004. After adjusting for the number of embryos transferred and class of embryos in a multivariate model, the best predictors were age (OR 0.918, 95% CI, 0.867-0.972 and AFC (OR 1.022, 95% CI, 0.992-1.053. Serum DHEAS levels were positively correlated with AFC (r = 0.098, P<0.039 and testosterone levels (r = 0.371, P<0.001, as well as the number of mature oocytes (r = 0.109, P<0.019; serum DHEAS levels were negatively correlated with age (r = -0.220, P<0.001, follicle-stimulating hormone (FSH, (r = -0.116, P<0.015 and sex hormone-binding globulin (SHBG, (r = -0.193, P<0.001.DHEAS concentration (in addition to the known factors of ovarian reserve does not predict clinical pregnancy in women with normal ovarian reserve who are undergoing ICSI.

  18. Air quality control in the ART laboratory is a major determinant of IVF success

    Science.gov (United States)

    Esteves, Sandro C; Bento, Fabiola C

    2016-01-01

    A recently published article described how a fertility center in the United States implemented air quality control to newly designed in vitro fertilization (IVF) laboratory.1 A highly-efficient air filtration was achieved by installing a centered system supplying filtered air to the IVF laboratory and related critical areas, combining air particulate and volatile organic compound (VOC) filtration. As a consequence, live birth rates were increased by improvements in air quality. This article highlights the key aspects of air contamination in the IVF context. The topic is important not only to IVF specialists but also to Andrologists due to the great number of male infertility patients referred to assisted reproductive technology (ART) treatments. The evidence is growing that laboratory air quality is paramount importance for improved IVF outcome. PMID:26585700

  19. Effect of insulin-like growth factor-1 during in vitro oocyte maturation and in vitro culture of bovine embryos Efeito do fator de crescimento semelhante à insulina-1 durante a maturação in vitro dos oócitos e cultivo in vitro de embriões bovinos

    Directory of Open Access Journals (Sweden)

    M.D. Quetglas

    2001-04-01

    Full Text Available The effects of insulin-like growth factor-I (IGF-I on in vitro maturation (IVM (experiment I and on in vitro embryo development (experiment II of bovine oocytes fertilized in vitro, were evaluated in terms of cleavage (CR, blastocyst (BR and hatching (HR rates. For IVM, immature cumulus-oocyte complexes were cultured in TCM-199 medium supplemented with Hepes, sodium bicarbonate, sodium pyruvate, additives, fetal calf serum (B-199 medium and gonadotropins (14 U/ml PMSG and 7 U/ml hCG. For embryo development, the oocytes/zygotes were cultured in B-199 medium with bovine oviduct epithelial cells in suspension under silicon oil. Treatments for in vitro culture conditions for both experiments were: 1- B-199 + 200 ng/ml IGF-I; 2- B-199 + 100 ng/ml IGF-I; 3- B-199 + 50 ng/ml IGF-I; 4- B-199 + 10 ng/ml IGF-I; 5- B-199 + 0 ng/ml IGF-I. All cultures were performed at 38.5ºC in 5% CO2 in air and the data were analyzed by chi-square test. In experiment I, there were no differences (P>0.05 among treatments for CR, BR or HR. In experiment II, the addition of IGF-I to the embryo culture medium (ECM resulted in a significant increase in CR while for BR and HR this effect was not observed. The addition of 200 ng/ml IGF-I to ECM increased CR (71.1% when compared to 100 ng/ml IGF-I (57.6% or control (56.7% groups, however, there were no differences when compared to 50 (69.4% or 10 ng/ml (73.1% groups. There was no beneficial effect of the addition of IGF-I in the IVM or ECM media on the in vitro development of embryos produced from oocytes matured and fertilized in vitro.Avaliaram-se o efeito do IGF-I na maturação in vitro (MIV (experimento I e no desenvolvimento embrionário (DE (experimento II de oócitos bovinos fecundados in vitro, quanto às taxas de clivagem (TC, de blastocistos (TB e de eclosão (TE. Para MIV, complexos cumulus-oócitos imaturos foram cultivados em meio TCM-199 suplementado com HEPES, bicarbonato e piruvato de sódio, aditivos, soro

  20. Cloning in cattle: from embryo splitting to somatic nuclear transfer.

    Science.gov (United States)

    Heyman, Y; Vignon, X; Chesné, P; Le Bourhis, D; Marchal, J; Renard, J P

    1998-01-01

    The ability to obtain genetically identical offspring in cattle (clones) is useful for research and for potential applications to breeding schemes. Experimental possibilities for generating such animals have evolved considerably in the last two decades. Embryo splitting has become a relatively simple technique but is limited to twinning. Embryonic nuclear transfer has improved and is associated with sexing to generate sets of clones despite a great variability of results between parent embryos. The factors of progress are reviewed here. Recently, somatic cells used as a source of nuclei in bovine nuclear transfer has been demonstrated. Here we present the results of the developmental potential of nuclei from skin and muscle cells.

  1. Comparison of conventional IVF versus ICSI in non-male factor, normoresponder patients

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    Maryam Eftekhar

    2012-01-01

    Full Text Available Background: Conventional IVF and ICSI are two common techniques to achieve fertilization. IVF has long been used for treatment of infertility, although it is not an effective treatment in severe male infertility. The use of ICSI has been expanded in severe male factor and fertilization failure after IVF cycle. In spite of the widespread use of ICSI in patients with non-male factor infertility, there is still little evidence to confirm its effectiveness in this population. Objective: To evaluate assisted reproductive technology outcomes between IVF and ICSI cycles in non-male factor, normoresponder patients. Materials and Methods: A total of 220 non-male factors, normoresponder patients who were indicated for ART were enrolled in this study. The patients received standard long GnRH agonist or GnRH antagonist protocols for ovarian stimulation and after oocytes retrieval, the patients were divided into two groups (IVF and ICSI groups. In IVF group (n=112, all of retrieved oocytes were treated by conventional IVF and in ICSI group (n=88, microinjection (ICSI was done on all of retrieved oocytes. Results: In IVF group, fertilization and implantation rates were significantly higher than ICSI group (66.22% and 16.67% in IVF group versus 57.46% and 11.17% in ICSI group, respectively. Chemical and clinical pregnancy rates were statistically higher in IVF group as compared with the ICSI group (42.9% vs. 27.3% and 35.7% vs. 21.5%, respectively. Conclusion: According to our study, the routine use of ICSI is not improved fertilization, implantation and chemical pregnancy rates and is not recommended in non-male factor, normozoospermic patients.

  2. 影响IVF-ET的因素研究

    Institute of Scientific and Technical Information of China (English)

    肖玉红

    2014-01-01

    目的探讨影响体外受精-胚胎移植(IVF-ET)的因素。方法收集我院自2012年10月~2013年4月我院行IVF-ET的患者的临床资料,应用多因素Logistic回归分析法进行分析。结果胚胎移植数、胚胎累积评分和受精率与妊娠结局正相关;年龄、促进性激素用量、基础卵泡雌刺激素值、移植出血量、人绒毛促性腺激素与妊娠结局负相关。结论胚胎移植数、胚胎累积评分、年龄、促进性激素用量、基础卵泡雌刺激素值、移植出血量、人绒毛促性腺激素和受精率等使影响妊娠结局的重要因素,应引起临床医生的重视。

  3. Effect of Administration of Single Dose GnRH Agonist in Luteal Phase on Outcome of ICSI-ET Cycles in Women with Previous History of IVF/ICSI Failure: A Randomized Controlled Trial

    Science.gov (United States)

    Zafardoust, Simin; Jeddi-Tehrani, Mahmood; Akhondi, Mohammad Mehdi; Sadeghi, Mohammad Reza; Kamali, Koroush; Mokhtar, Sara; Badehnoosh, Bita; Arjmand-Teymouri, Fatemeh; Fatemi, Farnaz; Mohammadzadeh, Afsaneh

    2015-01-01

    Background GnRH agonist administration in the luteal phase has been suggested to beneficially affect the outcome of intracytoplasmic sperm injection (ICSI) and embryo transfer (ET) cycles. This blind randomized controlled study evaluates the effect of GnRH (Gonadotropine Releasing Hormone) agonist administration on ICSI outcome in GnRH antagonist ovarian stimulation protocol in women with 2 or more previous IVF/ICSI-ET failures. Methods One hundred IVF failure women who underwent ICSI cycles and stimulated with GnRH antagonist ovarian stimulation protocol, were included in the study. Women were randomly assigned to intervention (received a single dose injection of GnRH agonist (0.1 mg of Decapeptil) subcutaneously 6 days after oocyte retrieval) and control (did not receive GnRH agonist) groups. Implantation and clinical pregnancy rates were the primary outcome measures. Results Although the age of women, the number of embryos transferred in the current cycle and the quality of the transferred embryos were similar in the two groups, there was a significantly higher rate of implantation (Mann Whitney test, p = 0.041) and pregnancy (32.6% vs. 12.5%, p = 0.030, OR = 3.3, 95%CI, 1.08 to 10.4) in the intervention group. Conclusion Our results suggested that, in addition to routine luteal phase support using progesterone, administration of 0.1 mg of Decapeptil 6 days after oocyte retrieval in women with previous history of 2 or more IVF/ICSI failures led to a significant improvement in implantation and pregnancy rates after ICSI following ovarian stimulation with GnRH antagonist protocol. PMID:25927026

  4. Follicular fluid insulin like growth factor-1 (FF IGF-1 is a biochemical marker of emb