Computational fluid dynamics modeling of momentum transport in rotating wall perfused bioreactor for cartilage tissue engineering.

Science.gov (United States)

Cinbiz, Mahmut N; Tığli, R Seda; Beşkardeş, Işil Gerçek; Gümüşderelioğlu, Menemşe; Colak, Uner

2010-11-01

In this study, computational fluid dynamics (CFD) analysis of a rotating-wall perfused-vessel (RWPV) bioreactor is performed to characterize the complex hydrodynamic environment for the simulation of cartilage development in RWPV bioreactor in the presence of tissue-engineered cartilage constructs, i.e., cell-chitosan scaffolds. Shear stress exerted on chitosan scaffolds in bioreactor was calculated for different rotational velocities in the range of 33-38 rpm. According to the calculations, the lateral and lower surfaces were exposed to 0.07926-0.11069 dyne/cm(2) and 0.05974-0.08345 dyne/cm(2), respectively, while upper surfaces of constructs were exposed to 0.09196-0.12847 dyne/cm(2). Results validate adequate hydrodynamic environment for scaffolds in RWPV bioreactor for cartilage tissue development which concludes the suitability of operational conditions of RWPV bioreactor. Copyright © 2010 Elsevier B.V. All rights reserved.

Predictive control of hollow-fiber bioreactors for the production of monoclonal antibodies.

Science.gov (United States)

Dowd, J E; Weber, I; Rodriguez, B; Piret, J M; Kwok, K E

1999-05-20

The selection of medium feed rates for perfusion bioreactors represents a challenge for process optimization, particularly in bioreactors that are sampled infrequently. When the present and immediate future of a bioprocess can be adequately described, predictive control can minimize deviations from set points in a manner that can maximize process consistency. Predictive control of perfusion hollow-fiber bioreactors was investigated in a series of hybridoma cell cultures that compared operator control to computer estimation of feed rates. Adaptive software routines were developed to estimate the current and predict the future glucose uptake and lactate production of the bioprocess at each sampling interval. The current and future glucose uptake rates were used to select the perfusion feed rate in a designed response to deviations from the set point values. The routines presented a graphical user interface through which the operator was able to view the up-to-date culture performance and assess the model description of the immediate future culture performance. In addition, fewer samples were taken in the computer-estimated cultures, reducing labor and analytical expense. The use of these predictive controller routines and the graphical user interface decreased the glucose and lactate concentration variances up to sevenfold, and antibody yields increased by 10% to 43%. Copyright 1999 John Wiley & Sons, Inc.

Perfusion based cell culture chips

DEFF Research Database (Denmark)

Heiskanen, Arto; Emnéus, Jenny; Dufva, Martin

2010-01-01

Performing cell culture in miniaturized perfusion chambers gives possibilities to experiment with cells under near in vivo like conditions. In contrast to traditional batch cultures, miniaturized perfusion systems provide precise control of medium composition, long term unattended cultures...... and tissue like structuring of the cultures. However, as this chapter illustrates, many issues remain to be identified regarding perfusion cell culture such as design, material choice and how to use these systems before they will be widespread amongst biomedical researchers....

The Influence of Bioreactor Geometry and the Mechanical Environment on Engineered Tissues

KAUST Repository

Osborne, J. M.; O’ Dea, R. D.; Whiteley, J. P.; Byrne, H. M.; Waters, S. L.

2010-01-01

A three phase model for the growth of a tissue construct within a perfusion bioreactor is examined. The cell population (and attendant extracellular matrix), culture medium, and porous scaffold are treated as distinct phases. The bioreactor system is represented by a two-dimensional channel containing a cell-seeded rigid porous scaffold (tissue construct), which is perfused with a culture medium. Through the prescription of appropriate functional forms for cell proliferation and extracellular matrix deposition rates, the model is used to compare the influence of cell density-, pressure-, and culture medium shear stress-regulated growth on the composition of the engineered tissue. The governing equations are derived in O'Dea et al. "A Three Phase Model for Tissue Construct Growth in a Perfusion Bioreactor," Math. Med. Biol., in which the long-wavelength limit was exploited to aid analysis; here, finite element methods are used to construct two-dimensional solutions to the governing equations and to investigate thoroughly their behavior. Comparison of the total tissue yield and averaged pressures, velocities, and shear stress demonstrates that quantitative agreement between the two-dimensional and long-wavelength approximation solutions is obtained for channel aspect ratios of order 10 -2 and that much of the qualitative behavior of the model is captured in the long-wavelength limit, even for relatively large channel aspect ratios. However, we demonstrate that in order to capture accurately the effect of mechanotransduction mechanisms on tissue construct growth, spatial effects in at least two dimensions must be included due to the inherent spatial variation of mechanical stimuli relevant to perfusion bioreactors, most notably, fluid shear stress, a feature not captured in the long-wavelength limit. Copyright © 2010 by ASME.

The Influence of Bioreactor Geometry and the Mechanical Environment on Engineered Tissues

KAUST Repository

Osborne, J. M.

2010-01-01

A three phase model for the growth of a tissue construct within a perfusion bioreactor is examined. The cell population (and attendant extracellular matrix), culture medium, and porous scaffold are treated as distinct phases. The bioreactor system is represented by a two-dimensional channel containing a cell-seeded rigid porous scaffold (tissue construct), which is perfused with a culture medium. Through the prescription of appropriate functional forms for cell proliferation and extracellular matrix deposition rates, the model is used to compare the influence of cell density-, pressure-, and culture medium shear stress-regulated growth on the composition of the engineered tissue. The governing equations are derived in O\\'Dea et al. "A Three Phase Model for Tissue Construct Growth in a Perfusion Bioreactor," Math. Med. Biol., in which the long-wavelength limit was exploited to aid analysis; here, finite element methods are used to construct two-dimensional solutions to the governing equations and to investigate thoroughly their behavior. Comparison of the total tissue yield and averaged pressures, velocities, and shear stress demonstrates that quantitative agreement between the two-dimensional and long-wavelength approximation solutions is obtained for channel aspect ratios of order 10 -2 and that much of the qualitative behavior of the model is captured in the long-wavelength limit, even for relatively large channel aspect ratios. However, we demonstrate that in order to capture accurately the effect of mechanotransduction mechanisms on tissue construct growth, spatial effects in at least two dimensions must be included due to the inherent spatial variation of mechanical stimuli relevant to perfusion bioreactors, most notably, fluid shear stress, a feature not captured in the long-wavelength limit. Copyright © 2010 by ASME.

An axial distribution of seeding, proliferation, and osteogenic differentiation of MC3T3-E1 cells and rat bone marrow-derived mesenchymal stem cells across a 3D Thai silk fibroin/gelatin/hydroxyapatite scaffold in a perfusion bioreactor

Energy Technology Data Exchange (ETDEWEB)

Sinlapabodin, Salita; Amornsudthiwat, Phakdee; Damrongsakkul, Siriporn; Kanokpanont, Sorada, E-mail: sorada.k@chula.ac.th

2016-01-01

In cell culture, a perfusion bioreactor provides effective transportation of nutrients, oxygen, and waste removal to and from the core of the scaffold. In addition, it provides mechanical stimuli for enhancing osteogenic differentiation. In this study, we used an axial distribution of cell numbers, alkaline phosphatase (ALP) enzyme activity, and calcium content across 4 cross-sections of 10 mm thick scaffold, made of Thai silk fibroin (SF)/gelatin (G)/hydroxyapatite (HA), as a tool to evaluate the suitable perfusion flow rate. These evaluations cover all cellular developmental phases starting from seeding, to proliferation, and later osteogenic differentiation. Mouse pre-osteoblastic MC3T3-E1 cell lines were used as a cell model during seeding and proliferation. The bioreactor seeded scaffold provided more uniform cell distribution across the scaffold compared to centrifugal and agitation seeding, while the overall number of adhered cells from bioreactor seeding was slightly lower than agitation seeding. The dynamic culture using 1 ml/min perfusion flow rate (initial shear stress of 0.1 dyn/cm{sup 2}) enabled statistically higher MC3T3-E1 proliferation, ALP activity, and calcium deposition than those observed in the static-culturing condition. However, the perfusion flow rate of 1 ml/min seemed not to be enough for enhancing ALP expression across all sections of the scaffold. Rat bone marrow derived stromal cells (rMSC) were used in the detachment test and osteogenic differentiation. It was found that perfusion flow rate of 5 ml/min caused statistically higher cell detachment than that of 1 and 3 ml/min. The perfusion flow rate of 3 ml/min gave the highest rMSC osteogenic differentiation on a SF/G/HA scaffold than other flow rates, as observed from the significantly highest number of ALP enzyme activity and the calcium content without any significant cell growth. In addition, all of these parameters were evenly distributed across all scaffold sections. - Highlights

Design modification and optimisation of the perfusion system of a tri-axial bioreactor for tissue engineering.

Science.gov (United States)

Hussein, Husnah; Williams, David J; Liu, Yang

2015-07-01

A systematic design of experiments (DOE) approach was used to optimize the perfusion process of a tri-axial bioreactor designed for translational tissue engineering exploiting mechanical stimuli and mechanotransduction. Four controllable design parameters affecting the perfusion process were identified in a cause-effect diagram as potential improvement opportunities. A screening process was used to separate out the factors that have the largest impact from the insignificant ones. DOE was employed to find the settings of the platen design, return tubing configuration and the elevation difference that minimise the load on the pump and variation in the perfusion process and improve the controllability of the perfusion pressures within the prescribed limits. DOE was very effective for gaining increased knowledge of the perfusion process and optimizing the process for improved functionality. It is hypothesized that the optimized perfusion system will result in improved biological performance and consistency.

3D perfusion bioreactor-activated porous granules on implant fixation and early bone formation in sheep.

Science.gov (United States)

Ding, Ming; Henriksen, Susan S; Martinetti, Roberta; Overgaard, Søren

2017-11-01

Early fixation of total joint arthroplasties is crucial for ensuring implant survival. An alternative bone graft material in revision surgery is needed to replace the current gold standard, allograft, seeing that the latter is associated with several disadvantages. The incubation of such a construct in a perfusion bioreactor has been shown to produce viable bone graft materials. This study aimed at producing larger amounts of viable bone graft material (hydroxyapatite 70% and β-tricalcium-phosphate 30%) in a novel perfusion bioreactor. The abilities of the bioreactor-activated graft material to induce early implant fixation were tested in a bilateral implant defect model in sheep, with allograft as the control group. Defects were bilaterally created in the distal femurs of the animals, and titanium implants were inserted. The concentric gaps around the implants were randomly filled with either allograft, granules, granules with bone marrow aspirate or bioreactor-activated graft material. Following an observation time of 6 weeks, early implant fixation and bone formation were assessed by micro-CT scanning, mechanical testing, and histomorphometry. Bone formations were seen in all groups, while no significant differences between groups were found regarding early implant fixation. The microarchitecture of the bone formed by the synthetic graft materials resembled that of allograft. Histomorphometry revealed that allograft induced significantly more bone and less fibrous tissue (p formation was observed in all groups, while the bioreactor-activated graft material did not reveal additional effects on early implant fixation comparable to allograft in this model. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 2465-2476, 2017. © 2016 Wiley Periodicals, Inc.

Perfusion cell culture decreases process and product heterogeneity in a head-to-head comparison with fed-batch.

Science.gov (United States)

Walther, Jason; Lu, Jiuyi; Hollenbach, Myles; Yu, Marcella; Hwang, Chris; McLarty, Jean; Brower, Kevin

2018-05-30

In this study, we compared the impacts of fed-batch and perfusion platforms on process and product attributes for IgG1- and IgG4-producing cell lines. A "plug-and-play" approach was applied to both platforms at bench scale, using commercially available basal and feed media, a standard feed strategy for fed-batch, and ATF filtration for perfusion. Product concentration in fed-batch was 2.5 times greater than perfusion, while average productivity in perfusion was 7.5 times greater than fed-batch. PCA revealed more variability in the cell environment and metabolism during the fed-batch run. LDH measurements showed that exposure of product to cell lysate was 7-10 times greater in fed-batch. Product analysis shows larger abundances of neutral species in perfusion, likely due to decreased bioreactor residence times and extracellular exposure. The IgG1 perfusion product also had higher purity and lower half-antibody. Glycosylation was similar across both culture modes. The first perfusion harvest slice for both product types showed different glycosylation than subsequent harvests, suggesting that product quality lags behind metabolism. In conclusion, process and product data indicate that intra-lot heterogeneity is decreased in perfusion cultures. Additional data and discussion is required to understand the developmental, clinical and commercial implications, and in what situations increased uniformity would be beneficial. This article is protected by copyright. All rights reserved.

Effects of Initial Seeding Density and Fluid Perfusion Rate on Formation of Tissue-Engineered Bone

OpenAIRE

GRAYSON, WARREN L.; BHUMIRATANA, SARINDR; CANNIZZARO, CHRISTOPHER; CHAO, P.-H. GRACE; LENNON, DONALD P.; CAPLAN, ARNOLD I.; VUNJAK-NOVAKOVIC, GORDANA

2008-01-01

We describe a novel bioreactor system for tissue engineering of bone that enables cultivation of up to six tissue constructs simultaneously, with direct perfusion and imaging capability. The bioreactor was used to investigate the relative effects of initial seeding density and medium perfusion rate on the growth and osteogenic differentiation patterns of bone marrow–derived human mesenchymal stem cells (hMSCs) cultured on three-dimensional scaffolds. Fully decellularized bovine trabecular bon...

Bioreactor-induced mesenchymal progenitor cell differentiation and elastic fiber assembly in engineered vascular tissues.

Science.gov (United States)

Lin, Shigang; Mequanint, Kibret

2017-09-01

In vitro maturation of engineered vascular tissues (EVT) requires the appropriate incorporation of smooth muscle cells (SMC) and extracellular matrix (ECM) components similar to native arteries. To this end, the aim of the current study was to fabricate 4mm inner diameter vascular tissues using mesenchymal progenitor cells seeded into tubular scaffolds. A dual-pump bioreactor operating either in perfusion or pulsatile perfusion mode was used to generate physiological-like stimuli to promote progenitor cell differentiation, extracellular elastin production, and tissue maturation. Our data demonstrated that pulsatile forces and perfusion of 3D tubular constructs from both the lumenal and ablumenal sides with culture media significantly improved tissue assembly, effectively inducing mesenchymal progenitor cell differentiation to SMCs with contemporaneous elastin production. With bioreactor cultivation, progenitor cells differentiated toward smooth muscle lineage characterized by the expression of smooth muscle (SM)-specific markers smooth muscle alpha actin (SM-α-actin) and smooth muscle myosin heavy chain (SM-MHC). More importantly, pulsatile perfusion bioreactor cultivation enhanced the synthesis of tropoelastin and its extracellular cross-linking into elastic fiber compared with static culture controls. Taken together, the current study demonstrated progenitor cell differentiation and vascular tissue assembly, and provides insights into elastin synthesis and assembly to fibers. Incorporation of elastin into engineered vascular tissues represents a critical design goal for both mechanical and biological functions. In the present study, we seeded porous tubular scaffolds with multipotent mesenchymal progenitor cells and cultured in dual-pump pulsatile perfusion bioreactor. Physiological-like stimuli generated by bioreactor not only induced mesenchymal progenitor cell differentiation to vascular smooth muscle lineage but also actively promoted elastin synthesis and

Bioreactor-Based Online Recovery of Human Progenitor Cells with Uncompromised Regenerative Potential: A Bone Tissue Engineering Perspective.

Directory of Open Access Journals (Sweden)

Maarten Sonnaert

Full Text Available The use of a 3D perfusion culture environment for stem cell expansion has been shown to be beneficial for maintenance of the original cell functionality but due to several system inherent characteristics such as the presence of extracellular matrix, the continued development and implementation of 3D perfusion bioreactor technologies is hampered. Therefore, this study developed a methodology for harvesting a progenitor cell population from a 3D open porous culture surface after expansion in a perfusion bioreactor and performed a functional characterization of the expanded cells. An initial screening showed collagenase to be the most interesting reagent to release the cells from the 3D culture surface as it resulted in high yields without compromising cell viability. Subsequently a Design of Experiment approach was used to obtain optimized 3D harvest conditions by assessing the interplay of flow rate, collagenase concentration and incubation time on the harvest efficiency, viability and single cell fraction. Cells that were recovered with the optimized harvest protocol, by perfusing a 880 U/ml collagenase solution for 7 hours at a flow rate of 4 ml/min, were thereafter functionally analyzed for their characteristics as expanded progenitor cell population. As both the in vitro tri-lineage differentiation capacity and the in vivo bone forming potential were maintained after 3D perfusion bioreactor expansion we concluded that the developed seeding, culture and harvest processes did not significantly compromise the viability and potency of the cells and can contribute to the future development of integrated bioprocesses for stem cell expansion.

Disposable Bioreactors for Plant Micropropagation and Mass Plant Cell Culture

Science.gov (United States)

Ducos, Jean-Paul; Terrier, Bénédicte; Courtois, Didier

Different types of bioreactors are used at Nestlé R&D Centre - Tours for mass propagation of selected plant varieties by somatic embryogenesis and for large scale culture of plants cells to produce metabolites or recombinant proteins. Recent studies have been directed to cut down the production costs of these two processes by developing disposable cell culture systems. Vegetative propagation of elite plant varieties is achieved through somatic embryogenesis in liquid medium. A pilot scale process has recently been set up for the industrial propagation of Coffea canephora (Robusta coffee). The current production capacity is 3.0 million embryos per year. The pre-germination of the embryos was previously conducted by temporary immersion in liquid medium in 10-L glass bioreactors. An improved process has been developed using a 10-L disposable bioreactor consisting of a bag containing a rigid plastic box ('Box-in-Bag' bioreactor), insuring, amongst other advantages, a higher light transmittance to the biomass due to its horizontal design. For large scale cell culture, two novel flexible plastic-based disposable bioreactors have been developed from 10 to 100 L working volumes, validated with several plant species ('Wave and Undertow' and 'Slug Bubble' bioreactors). The advantages and the limits of these new types of bioreactor are discussed, based mainly on our own experience on coffee somatic embryogenesis and mass cell culture of soya and tobacco.

Design and validation of a clinical-scale bioreactor for long-term isolated lung culture.

Science.gov (United States)

Charest, Jonathan M; Okamoto, Tatsuya; Kitano, Kentaro; Yasuda, Atsushi; Gilpin, Sarah E; Mathisen, Douglas J; Ott, Harald C

2015-06-01

The primary treatment for end-stage lung disease is lung transplantation. However, donor organ shortage remains a major barrier for many patients. In recent years, techniques for maintaining lungs ex vivo for evaluation and short-term (advance to more complex interventions for lung repair and regeneration, the need for a long-term organ culture system becomes apparent. Herein we describe a novel clinical scale bioreactor capable of maintaining functional porcine and human lungs for at least 72 h in isolated lung culture (ILC). The fully automated, computer controlled, sterile, closed circuit system enables physiologic pulsatile perfusion and negative pressure ventilation, while gas exchange function, and metabolism can be evaluated. Creation of this stable, biomimetic long-term culture environment will enable advanced interventions in both donor lungs and engineered grafts of human scale. Copyright © 2015 Elsevier Ltd. All rights reserved.

Biofabrication of customized bone grafts by combination of additive manufacturing and bioreactor knowhow.

Science.gov (United States)

Costa, Pedro F; Vaquette, Cédryck; Baldwin, Jeremy; Chhaya, Mohit; Gomes, Manuela E; Reis, Rui L; Theodoropoulos, Christina; Hutmacher, Dietmar W

2014-09-01

This study reports on an original concept of additive manufacturing for the fabrication of tissue engineered constructs (TEC), offering the possibility of concomitantly manufacturing a customized scaffold and a bioreactor chamber to any size and shape. As a proof of concept towards the development of anatomically relevant TECs, this concept was utilized for the design and fabrication of a highly porous sheep tibia scaffold around which a bioreactor chamber of similar shape was simultaneously built. The morphology of the bioreactor/scaffold device was investigated by micro-computed tomography and scanning electron microscopy confirming the porous architecture of the sheep tibiae as opposed to the non-porous nature of the bioreactor chamber. Additionally, this study demonstrates that both the shape, as well as the inner architecture of the device can significantly impact the perfusion of fluid within the scaffold architecture. Indeed, fluid flow modelling revealed that this was of significant importance for controlling the nutrition flow pattern within the scaffold and the bioreactor chamber, avoiding the formation of stagnant flow regions detrimental for in vitro tissue development. The bioreactor/scaffold device was dynamically seeded with human primary osteoblasts and cultured under bi-directional perfusion for two and six weeks. Primary human osteoblasts were observed homogenously distributed throughout the scaffold, and were viable for the six week culture period. This work demonstrates a novel application for additive manufacturing in the development of scaffolds and bioreactors. Given the intrinsic flexibility of the additive manufacturing technology platform developed, more complex culture systems can be fabricated which would contribute to the advances in customized and patient-specific tissue engineering strategies for a wide range of applications.

Biofabrication of customized bone grafts by combination of additive manufacturing and bioreactor knowhow

International Nuclear Information System (INIS)

Costa, Pedro F; Gomes, Manuela E; Reis, Rui L; Vaquette, Cédryck; Baldwin, Jeremy; Chhaya, Mohit; Theodoropoulos, Christina; Hutmacher, Dietmar W

2014-01-01

This study reports on an original concept of additive manufacturing for the fabrication of tissue engineered constructs (TEC), offering the possibility of concomitantly manufacturing a customized scaffold and a bioreactor chamber to any size and shape. As a proof of concept towards the development of anatomically relevant TECs, this concept was utilized for the design and fabrication of a highly porous sheep tibia scaffold around which a bioreactor chamber of similar shape was simultaneously built. The morphology of the bioreactor/scaffold device was investigated by micro-computed tomography and scanning electron microscopy confirming the porous architecture of the sheep tibiae as opposed to the non-porous nature of the bioreactor chamber. Additionally, this study demonstrates that both the shape, as well as the inner architecture of the device can significantly impact the perfusion of fluid within the scaffold architecture. Indeed, fluid flow modelling revealed that this was of significant importance for controlling the nutrition flow pattern within the scaffold and the bioreactor chamber, avoiding the formation of stagnant flow regions detrimental for in vitro tissue development. The bioreactor/scaffold device was dynamically seeded with human primary osteoblasts and cultured under bi-directional perfusion for two and six weeks. Primary human osteoblasts were observed homogenously distributed throughout the scaffold, and were viable for the six week culture period. This work demonstrates a novel application for additive manufacturing in the development of scaffolds and bioreactors. Given the intrinsic flexibility of the additive manufacturing technology platform developed, more complex culture systems can be fabricated which would contribute to the advances in customized and patient-specific tissue engineering strategies for a wide range of applications. (paper)

Flow perfusion culture of human mesenchymal stem cells on silicate-substituted tricalcium phosphate scaffolds

DEFF Research Database (Denmark)

Bjerre, Lea; Bünger, Cody E; Kassem, Moustapha

2008-01-01

Autologous bone grafts are currently the gold standard for treatment of large bone defects, but their availability is limited due to donor site morbidity. Different substitutes have been suggested to replace these grafts, and this study presents a bone tissue engineered alternative using silicate......-substituted tricalcium phosphate (Si-TCP) scaffolds seeded with human bone marrow-derived mesenchymal stem cells (hMSC). The cells were seeded onto the scaffolds and cultured either statically or in a perfusion bioreactor for up to 21 days and assessed for osteogenic differentiation by alkaline phosphatase activity...... assays and by quantitative real-time RT-PCR on bone markers. During culture, cells from the flow cultured constructs demonstrated improved proliferation and osteogenic differentiation verified by a more pronounced expression of several bone markers, e.g. alkaline phosphatase, osteopontin, Runx2, bone...

Miniature Bioreactor System for Long-Term Cell Culture

Science.gov (United States)

Gonda, Steve R.; Kleis, Stanley J.; Geffert, Sandara K.

2010-01-01

A prototype miniature bioreactor system is designed to serve as a laboratory benchtop cell-culturing system that minimizes the need for relatively expensive equipment and reagents and can be operated under computer control, thereby reducing the time and effort required of human investigators and reducing uncertainty in results. The system includes a bioreactor, a fluid-handling subsystem, a chamber wherein the bioreactor is maintained in a controlled atmosphere at a controlled temperature, and associated control subsystems. The system can be used to culture both anchorage-dependent and suspension cells, which can be either prokaryotic or eukaryotic. Cells can be cultured for extended periods of time in this system, and samples of cells can be extracted and analyzed at specified intervals. By integrating this system with one or more microanalytical instrument(s), one can construct a complete automated analytical system that can be tailored to perform one or more of a large variety of assays.

Perfusion-based three dimensional (3D) tissue engineering platform with integrated bioimpedance sensing

DEFF Research Database (Denmark)

Muhammad, Haseena Bashir; Canali, Chiara; Heiskanen, Arto

2014-01-01

We present an 8-channel bioreactor array with integrated bioimpedance sensors, which enables perfusion culture of cells seeded onto porous 3D scaffolds. Results show the capability of the system for monitoring cell proliferation within the scaffolds through a culture period of 19 days....

Hepatic Differentiation of Human Induced Pluripotent Stem Cells in a Perfused Three-Dimensional Multicompartment Bioreactor

Directory of Open Access Journals (Sweden)

Nora Freyer

2016-08-01

Full Text Available The hepatic differentiation of human induced pluripotent stem cells (hiPSC holds great potential for application in regenerative medicine, pharmacological drug screening, and toxicity testing. However, full maturation of hiPSC into functional hepatocytes has not yet been achieved. In this study, we investigated the potential of a dynamic three-dimensional (3D hollow fiber membrane bioreactor technology to improve the hepatic differentiation of hiPSC in comparison to static two-dimensional (2D cultures. A total of 100 × 106 hiPSC were seeded into each 3D bioreactor (n = 3. Differentiation into definitive endoderm (DE was induced by adding activin A, Wnt3a, and sodium butyrate to the culture medium. For further maturation, hepatocyte growth factor and oncostatin M were added. The same differentiation protocol was applied to hiPSC maintained in 2D cultures. Secretion of alpha-fetoprotein (AFP, a marker for DE, was significantly (p < 0.05 higher in 2D cultures, while secretion of albumin, a typical characteristic for mature hepatocytes, was higher after hepatic differentiation of hiPSC in 3D bioreactors. Functional analysis of multiple cytochrome P450 (CYP isoenzymes showed activity of CYP1A2, CYP2B6, and CYP3A4 in both groups, although at a lower level compared to primary human hepatocytes (PHH. CYP2B6 activities were significantly (p < 0.05 higher in 3D bioreactors compared with 2D cultures, which is in line with results from gene expression. Immunofluorescence staining showed that the majority of cells was positive for albumin, cytokeratin 18 (CK18, and hepatocyte nuclear factor 4-alpha (HNF4A at the end of the differentiation process. In addition, cytokeratin 19 (CK19 staining revealed the formation of bile duct-like structures in 3D bioreactors similar to native liver tissue. The results indicate a better maturation of hiPSC in the 3D bioreactor system compared to 2D cultures and emphasize the potential of dynamic 3D culture

Bioreactor perfusion system for the long-term maintenance of tissue-engineered skeletal muscle organoids

Science.gov (United States)

Chromiak, J. A.; Shansky, J.; Perrone, C.; Vandenburgh, H. H.

1998-01-01

Three-dimensional skeletal muscle organ-like structures (organoids) formed in tissue culture by fusion of proliferating myoblasts into parallel networks of long, unbranched myofibers provide an in vivo-like model for examining the effects of growth factors, tension, and space flight on muscle cell growth and metabolism. To determine the feasibility of maintaining either avian or mammalian muscle organoids in a commercial perfusion bioreactor system, we measured metabolism, protein turnover. and autocrine/paracrine growth factor release rates. Medium glucose was metabolized at a constant rate in both low-serum- and serum-free media for up to 30 d. Total organoid noncollagenous protein and DNA content decreased approximately 22-28% (P skeletal muscle growth factors prostaglandin F2alpha (PGF2alpha) and insulin-like growth factor-1 (IGF-1) could be measured accurately in collected media fractions, even after storage at 37 degrees C for up to 10 d. In contrast, creatine kinase activity (a marker of cell damage) in collected media fractions was unreliable. These results provide initial benchmarks for long-term ex vivo studies of tissue-engineered skeletal muscle.

Maximizing neotissue growth kinetics in a perfusion bioreactor: An in silico strategy using model reduction and Bayesian optimization.

Science.gov (United States)

Mehrian, Mohammad; Guyot, Yann; Papantoniou, Ioannis; Olofsson, Simon; Sonnaert, Maarten; Misener, Ruth; Geris, Liesbet

2018-03-01

In regenerative medicine, computer models describing bioreactor processes can assist in designing optimal process conditions leading to robust and economically viable products. In this study, we started from a (3D) mechanistic model describing the growth of neotissue, comprised of cells, and extracellular matrix, in a perfusion bioreactor set-up influenced by the scaffold geometry, flow-induced shear stress, and a number of metabolic factors. Subsequently, we applied model reduction by reformulating the problem from a set of partial differential equations into a set of ordinary differential equations. Comparing the reduced model results to the mechanistic model results and to dedicated experimental results assesses the reduction step quality. The obtained homogenized model is 10 5 fold faster than the 3D version, allowing the application of rigorous optimization techniques. Bayesian optimization was applied to find the medium refreshment regime in terms of frequency and percentage of medium replaced that would maximize neotissue growth kinetics during 21 days of culture. The simulation results indicated that maximum neotissue growth will occur for a high frequency and medium replacement percentage, a finding that is corroborated by reports in the literature. This study demonstrates an in silico strategy for bioprocess optimization paying particular attention to the reduction of the associated computational cost. © 2017 Wiley Periodicals, Inc.

Dynamic 3D culture promotes spontaneous embryonic stem cell differentiation in vitro.

Science.gov (United States)

Gerlach, Jörg C; Hout, Mariah; Edsbagge, Josefina; Björquist, Petter; Lübberstedt, Marc; Miki, Toshio; Stachelscheid, Harald; Schmelzer, Eva; Schatten, Gerald; Zeilinger, Katrin

2010-02-01

Spontaneous in vitro differentiation of mouse embryonic stem cells (mESC) is promoted by a dynamic, three-dimensional (3D), tissue-density perfusion technique with continuous medium perfusion and exchange in a novel four-compartment, interwoven capillary bioreactor. We compared ectodermal, endodermal, and mesodermal immunoreactive tissue structures formed by mESC at culture day 10 with mouse fetal tissue development at gestational day E9.5. The results show that the bioreactor cultures more closely resemble mouse fetal tissue development at gestational day E9.5 than control mESC cultured in Petri dishes.

Bioreactor design for successive culture of anchorage-dependent cells operated in an automated manner.

Science.gov (United States)

Kino-Oka, Masahiro; Ogawa, Natsuki; Umegaki, Ryota; Taya, Masahito

2005-01-01

A novel bioreactor system was designed to perform a series of batchwise cultures of anchorage-dependent cells by means of automated operations of medium change and passage for cell transfer. The experimental data on contamination frequency ensured the biological cleanliness in the bioreactor system, which facilitated the operations in a closed environment, as compared with that in flask culture system with manual handlings. In addition, the tools for growth prediction (based on growth kinetics) and real-time growth monitoring by measurement of medium components (based on small-volume analyzing machinery) were installed into the bioreactor system to schedule the operations of medium change and passage and to confirm that culture proceeds as scheduled, respectively. The successive culture of anchorage-dependent cells was conducted with the bioreactor running in an automated way. The automated bioreactor gave a successful culture performance with fair accordance to preset scheduling based on the information in the latest subculture, realizing 79- fold cell expansion for 169 h. In addition, the correlation factor between experimental data and scheduled values through the bioreactor performance was 0.998. It was concluded that the proposed bioreactor with the integration of the prediction and monitoring tools could offer a feasible system for the manufacturing process of cultured tissue products.

Method and Apparatus for a Miniature Bioreactor System for Long-Term Cell Culture

Science.gov (United States)

Kleis, Stanley J. (Inventor); Geffert, Sandra K. (Inventor); Gonda, Steve R. (Inventor)

2015-01-01

A bioreactor and method that permits continuous and simultaneous short, moderate, or long term cell culturing of one or more cell types or tissue in a laminar flow configuration is disclosed, where the bioreactor supports at least two laminar flow zones, which are isolated by laminar flow without the need for physical barriers between the zones. The bioreactors of this invention are ideally suited for studying short, moderate and long term studies of cell cultures and the response of cell cultures to one or more stressors such as pharmaceuticals, hypoxia, pathogens, or any other stressor. The bioreactors of this invention are also ideally suited for short, moderate or long term cell culturing with periodic cell harvesting and/or medium processing for secreted cellular components.

Differentiation of cartilaginous anlage in entire embryonic mouse limbs cultured in a rotating bioreactor.

Science.gov (United States)

Duke, P.; Oakley, C.; Montufar-Solis, D.

The embryonic mammalian limb is sensitive both in vivo and in vitro to changes in gravitational force. Hypergravity of centrifugation and microgravity of space decreased size of elements due to precocious or delayed chondrogenesis respectively. In recapitulating spaceflight experiments, premetatarsals were cultured in suspension in a low stress, low sheer rotating bioreactor, and found to be shorter than those cultured in standard culture dishes, and cartilage development was delayed. This study only measured length of the metatarsals, and did not account for possible changes in width and/or in form of the skeletal elements. Shorter cartilage elements in limbbuds cultured in the bioreactor may be due to the ability of the system to reproduce a more in vivo 3D shape than traditional organ cultures. Tissues subjected to traditional organ cultures become flattened by their own weight, attachment to the filter, and restrictions imposed by nutrient diffusion. The purpose of the current experiment was to determine if entire limb buds could be successfully cultured in the bioreactor, and to compare the effects on 3D shape with that of culturing in a culture dish system. Fore and hind limbs from E11-E13 ICR mouse embryos were placed either in the bioreactor, in Trowell culture, or fixed as controls. Limbbuds were cultured for six days, fixed, and processed either as whole mounts or embedded for histology. Qualitative analysis revealed that the Trowell culture specimens were flattened, while bioreactor culture specimens had a more in vivo-like 3D limb shape. Sections of limbbuds from both types of cultures had excellent cartilage differentiation, with apparently more cell maturation, and hypertrophy in the specimens cultured in the bioreactor. Morphometric quantitation of the cartilaginous elements for comparisons of the two culture systems was complicated due to some limb buds fusing together during culture. This problem was especially noticeable in the younger limbs, and

Visualizing feasible operating ranges within tissue engineering systems using a "windows of operation" approach: a perfusion-scaffold bioreactor case study.

Science.gov (United States)

McCoy, Ryan J; O'Brien, Fergal J

2012-12-01

Tissue engineering approaches to developing functional substitutes are often highly complex, multivariate systems where many aspects of the biomaterials, bio-regulatory factors or cell sources may be controlled in an effort to enhance tissue formation. Furthermore, success is based on multiple performance criteria reflecting both the quantity and quality of the tissue produced. Managing the trade-offs between different performance criteria is a challenge. A "windows of operation" tool that graphically represents feasible operating spaces to achieve user-defined levels of performance has previously been described by researchers in the bio-processing industry. This paper demonstrates the value of "windows of operation" to the tissue engineering field using a perfusion-scaffold bioreactor system as a case study. In our laboratory, perfusion bioreactor systems are utilized in the context of bone tissue engineering to enhance the osteogenic differentiation of cell-seeded scaffolds. A key challenge of such perfusion bioreactor systems is to maximize the induction of osteogenesis but minimize cell detachment from the scaffold. Two key operating variables that influence these performance criteria are the mean scaffold pore size and flow-rate. Using cyclooxygenase-2 and osteopontin gene expression levels as surrogate indicators of osteogenesis, we employed the "windows of operation" methodology to rapidly identify feasible operating ranges for the mean scaffold pore size and flow-rate that achieved user-defined levels of performance for cell detachment and differentiation. Incorporation of such tools into the tissue engineer's armory will hopefully yield a greater understanding of the highly complex systems used and help aid decision making in future translation of products from the bench top to the market place. Copyright © 2012 Wiley Periodicals, Inc.

Cell culture experiments planned for the space bioreactor

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Morrison, Dennis R.; Cross, John H.

1987-01-01

Culturing of cells in a pilot-scale bioreactor remains to be done in microgravity. An approach is presented based on several studies of cell culture systems. Previous and current cell culture research in microgravity which is specifically directed towards development of a space bioprocess is described. Cell culture experiments planned for a microgravity sciences mission are described in abstract form.

Metabolic and Kinetic analyses of influenza production in perfusion HEK293 cell culture

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Lohr Verena

2011-09-01

Full Text Available Abstract Background Cell culture-based production of influenza vaccine remains an attractive alternative to egg-based production. Short response time and high production yields are the key success factors for the broader adoption of cell culture technology for industrial manufacturing of pandemic and seasonal influenza vaccines. Recently, HEK293SF cells have been successfully used to produce influenza viruses, achieving hemagglutinin (HA and infectious viral particle (IVP titers in the highest ranges reported to date. In the same study, it was suggested that beyond 4 × 106 cells/mL, viral production was limited by a lack of nutrients or an accumulation of toxic products. Results To further improve viral titers at high cell densities, perfusion culture mode was evaluated. Productivities of both perfusion and batch culture modes were compared at an infection cell density of 6 × 106 cells/mL. The metabolism, including glycolysis, glutaminolysis and amino acids utilization as well as physiological indicators such as viability and apoptosis were extensively documented for the two modes of culture before and after viral infection to identify potential metabolic limitations. A 3 L bioreactor with a perfusion rate of 0.5 vol/day allowed us to reach maximal titers of 3.3 × 1011 IVP/mL and 4.0 logHA units/mL, corresponding to a total production of 1.0 × 1015 IVP and 7.8 logHA units after 3 days post-infection. Overall, perfusion mode titers were higher by almost one order of magnitude over the batch culture mode of production. This improvement was associated with an activation of the cell metabolism as seen by a 1.5-fold and 4-fold higher consumption rates of glucose and glutamine respectively. A shift in the viral production kinetics was also observed leading to an accumulation of more viable cells with a higher specific production and causing an increase in the total volumetric production of infectious influenza particles. Conclusions These results

Hydrogel/poly-dimethylsiloxane hybrid bioreactor facilitating 3D cell culturing

NARCIS (Netherlands)

Schurink, B.; Luttge, R.

2013-01-01

The authors present a hydrogel/poly-dimethylsiloxane (PDMS) hybrid bioreactor. The bioreactor enables a low shear stress 3D culture by integrating a hydrogel as a barrier into a PDMS casing. The use of PDMS allows the reversible adhesion of the device to a commercially available microelectrode

High-throughput miniaturized bioreactors for cell culture process development: reproducibility, scalability, and control.

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Rameez, Shahid; Mostafa, Sigma S; Miller, Christopher; Shukla, Abhinav A

2014-01-01

Decreasing the timeframe for cell culture process development has been a key goal toward accelerating biopharmaceutical development. Advanced Microscale Bioreactors (ambr™) is an automated micro-bioreactor system with miniature single-use bioreactors with a 10-15 mL working volume controlled by an automated workstation. This system was compared to conventional bioreactor systems in terms of its performance for the production of a monoclonal antibody in a recombinant Chinese Hamster Ovary cell line. The miniaturized bioreactor system was found to produce cell culture profiles that matched across scales to 3 L, 15 L, and 200 L stirred tank bioreactors. The processes used in this article involve complex feed formulations, perturbations, and strict process control within the design space, which are in-line with processes used for commercial scale manufacturing of biopharmaceuticals. Changes to important process parameters in ambr™ resulted in predictable cell growth, viability and titer changes, which were in good agreement to data from the conventional larger scale bioreactors. ambr™ was found to successfully reproduce variations in temperature, dissolved oxygen (DO), and pH conditions similar to the larger bioreactor systems. Additionally, the miniature bioreactors were found to react well to perturbations in pH and DO through adjustments to the Proportional and Integral control loop. The data presented here demonstrates the utility of the ambr™ system as a high throughput system for cell culture process development. © 2014 American Institute of Chemical Engineers.

Validation of an in vitro 3D bone culture model with perfused and mechanically stressed ceramic scaffold

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G Bouet

2015-05-01

Full Text Available An engineered three dimensional (3D in vitro cell culture system was designed with the goal of inducing and controlling in vitro osteogenesis in a reproducible manner under conditions more similar to the in vivo bone microenvironment than traditional two-dimensional (2D models. This bioreactor allows efficient mechanical loading and perfusion of an original cubic calcium phosphate bioceramic of highly controlled composition and structure. This bioceramic comprises an internal portion containing homogeneously interconnected macropores surrounded by a dense layer, which minimises fluid flow bypass around the scaffold. This dense and flat layer permits the application of a homogeneous loading on the bioceramic while also enhancing its mechanical strength. Numerical modelling of constraints shows that the system provides direct mechanical stimulation of cells within the scaffold. Experimental results establish that under perfusion at a steady flow of 2 µL/min, corresponding to 3 ≤ Medium velocity ≤ 23 µm/s, mouse calvarial cells grow and differentiate as osteoblasts in a reproducible manner, and lay down a mineralised matrix. Moreover, cells respond to mechanical loading by increasing C-fos expression, which demonstrates the effective mechanical stimulation of the culture within the scaffold. In summary, we provide a “proof-of-concept” for osteoblastic cell culture in a controlled 3D culture system under perfusion and mechanical loading. This model will be a tool to analyse bone cell functions in vivo, and will provide a bench testing system for the clinical assessment of bioactive bone-targeting molecules under load.

Differentiation of cartilaginous anlagen in entire embryonic mouse limbs cultured in a rotating bioreactor

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Montufar-Solis, D.; Oakley, C. R.; Jefferson, Y.; Duke, P. J.

2003-10-01

Mechanisms involved in development of the embryonic limb have remained the same throughout eons of genetic and environmental evolution under Earth gravity (lg). During the spaceflight era it has been of interest to explore the ancient theory that form of the skeleton develops in response to gravity, and that changes in gravitational forces can change the developmental pattern of the limb. This has been shown in vivo and in vitro, allowing the hypergravity of centrifugation and microgravity of space to be used as tools to increase our knowledge of limb development. In recapitulations of spaceflight experiments, premetatarsals were cultured in suspension in a bioreactor, and found to be shorter and less differentiated than those cultured in standard culture dishes. This study only measured length of the metatarsals, and did not account for possible changes due to the skeletal elements having a more in vivo 3D shape while in suspension vs. flattened tissues compressed by their own weight. A culture system with an outcome closer to in vivo and that supports growth of younger limb buds than traditional systems will allow studies of early Hox gene expression, and contribute to the understanding of very early stages of development. The purpose of the current experiment was to determine if entire limb buds could be cultured in the bioreactor, and to compare the growth and differentiation with that of culturing in a culture dish system. Fore and hind limbs from E11-E13 ICR mouse embryos were cultured for six days, either in the bioreactor or in center-well organ culture dishes, fixed, and embedded for histology. E13 specimens grown in culture dishes were flat, while bioreactor culture specimens had a more in vivo-like 3D limb shape. Sections showed excellent cartilage differentiation in both culture systems, with more cell maturation, and hypertrophy in the specimens cultured in the bioreactor. Younger limb buds fused together during culture, so an additional set of El 1

Detachably assembled microfluidic device for perfusion culture and post-culture analysis of a spheroid array.

Science.gov (United States)

Sakai, Yusuke; Hattori, Koji; Yanagawa, Fumiki; Sugiura, Shinji; Kanamori, Toshiyuki; Nakazawa, Kohji

2014-07-01

Microfluidic devices permit perfusion culture of three-dimensional (3D) tissue, mimicking the flow of blood in vascularized 3D tissue in our body. Here, we report a microfluidic device composed of a two-part microfluidic chamber chip and multi-microwell array chip able to be disassembled at the culture endpoint. Within the microfluidic chamber, an array of 3D tissue aggregates (spheroids) can be formed and cultured under perfusion. Subsequently, detailed post-culture analysis of the spheroids collected from the disassembled device can be performed. This device facilitates uniform spheroid formation, growth analysis in a high-throughput format, controlled proliferation via perfusion flow rate, and post-culture analysis of spheroids. We used the device to culture spheroids of human hepatocellular carcinoma (HepG2) cells under two controlled perfusion flow rates. HepG2 spheroids exhibited greater cell growth at higher perfusion flow rates than at lower perfusion flow rates, and exhibited different metabolic activity and mRNA and protein expression under the different flow rate conditions. These results show the potential of perfusion culture to precisely control the culture environment in microfluidic devices. The construction of spheroid array chambers allows multiple culture conditions to be tested simultaneously, with potential applications in toxicity and drug screening. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Model-Based Optimization of Scaffold Geometry and Operating Conditions of Radial Flow Packed-Bed Bioreactors for Therapeutic Applications

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Danilo Donato

2014-01-01

Full Text Available Radial flow perfusion of cell-seeded hollow cylindrical porous scaffolds may overcome the transport limitations of pure diffusion and direct axial perfusion in the realization of bioengineered substitutes of failing or missing tissues. Little has been reported on the optimization criteria of such bioreactors. A steady-state model was developed, combining convective and dispersive transport of dissolved oxygen with Michaelis-Menten cellular consumption kinetics. Dimensional analysis was used to combine more effectively geometric and operational variables in the dimensionless groups determining bioreactor performance. The effectiveness of cell oxygenation was expressed in terms of non-hypoxic fractional construct volume. The model permits the optimization of the geometry of hollow cylindrical constructs, and direction and magnitude of perfusion flow, to ensure cell oxygenation and culture at controlled oxygen concentration profiles. This may help engineer tissues suitable for therapeutic and drug screening purposes.

Bioreactor principles

Science.gov (United States)

2001-01-01

Cells cultured on Earth (left) typically settle quickly on the bottom of culture vessels due to gravity. In microgravity (right), cells remain suspended and aggregate to form three-dimensional tissue. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. The Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. Due to their high level of cellular organization and specialization, samples constructed in the bioreactor more closely resemble the original tumor or tissue found in the body. The work is sponsored by NASA's Office of Biological and Physical Research. The bioreactor is managed by the Biotechnology Cell Science Program at NASA's Johnson Space Center (JSC). NASA-sponsored bioreactor research has been instrumental in helping scientists to better understand normal and cancerous tissue development. In cooperation with the medical community, the bioreactor design is being used to prepare better models of human colon, prostate, breast and ovarian tumors. Cartilage, bone marrow, heart muscle, skeletal muscle, pancreatic islet cells, liver and kidney are just a few of the normal tissues being cultured in rotating bioreactors by investigators.

In vitro evaluation of a novel bioreactor based on an integral oxygenator and a spirally wound nonwoven polyester matrix for hepatocyte culture as small aggregates

NARCIS (Netherlands)

Flendrig, L. M.; la Soe, J. W.; Jörning, G. G.; Steenbeek, A.; Karlsen, O. T.; Bovée, W. M.; Ladiges, N. C.; te Velde, A. A.; Chamuleau, R. A.

1997-01-01

BACKGROUND/AIMS: The development of custom-made bioreactors for use as a bioartificial liver (BAL) is considered to be one of the last challenges on the road to successful temporary extracorporeal liver support therapy. We devised a novel bioreactor (patent pending) which allows individual perfusion

Spaceflight bioreactor studies of cells and tissues.

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Freed, Lisa E; Vunjak-Novakovic, Gordana

2002-01-01

well-being (loss of muscle and skeletal tissues [15-17]) and gene- and cell-level responses to the mechanical environment [13,14,18]. All five of the spaceflight bioreactor studies described above utilized three-dimensional cell culture systems in which the cells were associated with biodegradable polymer scaffolds [17], collagen gel [16], or microcarrier beads [13-15,18] in order to promote the expression of differentiated cell function. In four of the five spaceflight bioreactor studies [15-18], cells were cultured in perfused vessels (cartridges or rotating bioreactors) within recirculating loops designed to maintain medium composition within target ranges by a combination of gas exchange and fresh medium supply. Future spaceflight studies of cells and tissues are likely to involve a three-dimensional culture system, to promote cellular differentiation, and perfusion with or without rotation, to provide a gravity-independent mechanism for fluid mixing and mass transport. Previous spaceflight studies have guided the ongoing development of NASA flight hardware for the ISS (e.g. the EDU-2 and the CCU). This next generation of hardware will have extended operational capabilities including on-line microscopy, in-line sensors for the monitoring and control of metabolic parameters, modular design for replicate cultures, and, perhaps most importantly of all, compatibility with the ISS centrifuge. The latter will permit in-flight, 1 g control cultures, and thereby allow the experimental variable to be gravity itself rather than the more general "spaceflight environment". Technical limitations of spaceflight studies (e.g. allowable size, mass, and power) continue to motivate a creative approach to system design and to result in "spin-off" technologies (e.g. the STLV) for ground-based cell and tissue culture research. The increasing scientific and medical relevance of this work is evidenced by the growing number of publications in which advanced bioreactors are used for in

Evaluation of a Multi-Parameter Sensor for Automated, Continuous Cell Culture Monitoring in Bioreactors

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Pappas, D.; Jeevarajan, A.; Anderson, M. M.

2004-01-01

Compact and automated sensors are desired for assessing the health of cell cultures in biotechnology experiments in microgravity. Measurement of cell culture medium allows for the optirn.jzation of culture conditions on orbit to maximize cell growth and minimize unnecessary exchange of medium. While several discrete sensors exist to measure culture health, a multi-parameter sensor would simplify the experimental apparatus. One such sensor, the Paratrend 7, consists of three optical fibers for measuring pH, dissolved oxygen (p02), dissolved carbon dioxide (pC02) , and a thermocouple to measure temperature. The sensor bundle was designed for intra-arterial placement in clinical patients, and potentially can be used in NASA's Space Shuttle and International Space Station biotechnology program bioreactors. Methods: A Paratrend 7 sensor was placed at the outlet of a rotating-wall perfused vessel bioreactor system inoculated with BHK-21 (baby hamster kidney) cells. Cell culture medium (GTSF-2, composed of 40% minimum essential medium, 60% L-15 Leibovitz medium) was manually measured using a bench top blood gas analyzer (BGA, Ciba-Corning). Results: A Paratrend 7 sensor was used over a long-term (>120 day) cell culture experiment. The sensor was able to track changes in cell medium pH, p02, and pC02 due to the consumption of nutrients by the BHK-21. When compared to manually obtained BGA measurements, the sensor had good agreement for pH, p02, and pC02 with bias [and precision] of 0.02 [0.15], 1 mm Hg [18 mm Hg], and -4.0 mm Hg [8.0 mm Hg] respectively. The Paratrend oxygen sensor was recalibrated (offset) periodically due to drift. The bias for the raw (no offset or recalibration) oxygen measurements was 42 mm Hg [38 mm Hg]. The measured response (rise) time of the sensor was 20 +/- 4s for pH, 81 +/- 53s for pC02, 51 +/- 20s for p02. For long-term cell culture measurements, these response times are more than adequate. Based on these findings , the Paratrend sensor could

Development of a microfluidic perfusion 3D cell culture system

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Park, D. H.; Jeon, H. J.; Kim, M. J.; Nguyen, X. D.; Morten, K.; Go, J. S.

2018-04-01

Recently, 3-dimensional in vitro cell cultures have gained much attention in biomedical sciences because of the closer relevance between in vitro cell cultures and in vivo environments. This paper presents a microfluidic perfusion 3D cell culture system with consistent control of long-term culture conditions to mimic an in vivo microenvironment. It consists of two sudden expansion reservoirs to trap incoming air bubbles, gradient generators to provide a linear concentration, and microchannel mixers. Specifically, the air bubbles disturb a flow in the microfluidic channel resulting in the instability of the perfusion cell culture conditions. For long-term stable operation, the sudden expansion reservoir is designed to trap air bubbles by using buoyancy before they enter the culture system. The performance of the developed microfluidic perfusion 3D cell culture system was examined experimentally and compared with analytical results. Finally, it was applied to test the cytotoxicity of cells infected with Ewing’s sarcoma. Cell death was observed for different concentrations of H2O2. For future work, the developed microfluidic perfusion 3D cell culture system can be used to examine the behavior of cells treated with various drugs and concentrations for high-throughput drug screening.

Developing a Customized Perfusion Bioreactor Prototype with Controlled Positional Variability in Oxygen Partial Pressure for Bone and Cartilage Tissue Engineering.

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Lee, Poh Soo; Eckert, Hagen; Hess, Ricarda; Gelinsky, Michael; Rancourt, Derrick; Krawetz, Roman; Cuniberti, Gianaurelio; Scharnweber, Dieter

2017-05-01

Skeletal development is a multistep process that involves the complex interplay of multiple cell types at different stages of development. Besides biochemical and physical cues, oxygen tension also plays a pivotal role in influencing cell fate during skeletal development. At physiological conditions, bone cells generally reside in a relatively oxygenated environment whereas chondrocytes reside in a hypoxic environment. However, it is technically challenging to achieve such defined, yet diverse oxygen distribution on traditional in vitro cultivation platforms. Instead, engineered osteochondral constructs are commonly cultivated in a homogeneous, stable environment. In this study, we describe a customized perfusion bioreactor having stable positional variability in oxygen tension at defined regions. Further, engineered collagen constructs were coaxed into adopting the shape and dimensions of defined cultivation platforms that were precasted in 1.5% agarose bedding. After cultivating murine embryonic stem cells that were embedded in collagen constructs for 50 days, mineralized constructs of specific dimensions and a stable structural integrity were achieved. The end-products, specifically constructs cultivated without chondroitin sulfate A (CSA), showed a significant increase in mechanical stiffness compared with their initial gel-like constructs. More importantly, the localization of osteochondral cell types was specific and corresponded to the oxygen tension gradient generated in the bioreactor. In addition, CSA in complementary with low oxygen tension was also found to be a potent inducer of chondrogenesis in this system. In summary, we have demonstrated a customized perfusion bioreactor prototype that is capable of generating a more dynamic, yet specific cultivation environment that could support propagation of multiple osteochondral lineages within a single engineered construct in vitro. Our system opens up new possibilities for in vitro research on human

Antioxidant compounds in Salvia officinalis L. shoot and hairy root cultures in the nutrient sprinkle bioreactor

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Izabela Grzegorczyk

2011-01-01

Full Text Available The study focused on the production of compounds with antioxidant activity in hairy root and shoot cultures of Salvia officinalis grown in laboratory-scale sprinkle nutrient bioreactors. HPLC analysis showed that production of rosmarinic acid in transformed roots (34.65 ±1.07 mg l-1 was higher that in shoot culture (26.24 ±0.48 mg l-1. In the latter diterpenoids: carnosic acid (1.74 ±0.02 mg l-1 and carnosol (1.34 ±0.01 mg l-1 were also found. Biomass accumulation after a growth period in the bioreactor was also studied. An 18-fold increase in hairy root biomass was recorded after 40 days of culture. In sage shoot culture, biomass increased 43 times after 21 days of bioreactor run. The current operating conditions of the bioreactor were not suitable for the propagation of Salvia officinalis mainly due to the hyperhydricity problem of leaves and stems.

Bioreactors for high cell density and continuous multi-stage cultivations: options for process intensification in cell culture-based viral vaccine production.

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Tapia, Felipe; Vázquez-Ramírez, Daniel; Genzel, Yvonne; Reichl, Udo

2016-03-01

With an increasing demand for efficacious, safe, and affordable vaccines for human and animal use, process intensification in cell culture-based viral vaccine production demands advanced process strategies to overcome the limitations of conventional batch cultivations. However, the use of fed-batch, perfusion, or continuous modes to drive processes at high cell density (HCD) and overextended operating times has so far been little explored in large-scale viral vaccine manufacturing. Also, possible reductions in cell-specific virus yields for HCD cultivations have been reported frequently. Taking into account that vaccine production is one of the most heavily regulated industries in the pharmaceutical sector with tough margins to meet, it is understandable that process intensification is being considered by both academia and industry as a next step toward more efficient viral vaccine production processes only recently. Compared to conventional batch processes, fed-batch and perfusion strategies could result in ten to a hundred times higher product yields. Both cultivation strategies can be implemented to achieve cell concentrations exceeding 10(7) cells/mL or even 10(8) cells/mL, while keeping low levels of metabolites that potentially inhibit cell growth and virus replication. The trend towards HCD processes is supported by development of GMP-compliant cultivation platforms, i.e., acoustic settlers, hollow fiber bioreactors, and hollow fiber-based perfusion systems including tangential flow filtration (TFF) or alternating tangential flow (ATF) technologies. In this review, these process modes are discussed in detail and compared with conventional batch processes based on productivity indicators such as space-time yield, cell concentration, and product titers. In addition, options for the production of viral vaccines in continuous multi-stage bioreactors such as two- and three-stage systems are addressed. While such systems have shown similar virus titers compared to

A carbon dioxide stripping model for mammalian cell culture in manufacturing scale bioreactors.

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Xing, Zizhuo; Lewis, Amanda M; Borys, Michael C; Li, Zheng Jian

2017-06-01

Control of carbon dioxide within the optimum range is important in mammalian bioprocesses at the manufacturing scale in order to ensure robust cell growth, high protein yields, and consistent quality attributes. The majority of bioprocess development work is done in laboratory bioreactors, in which carbon dioxide levels are more easily controlled. Some challenges in carbon dioxide control can present themselves when cell culture processes are scaled up, because carbon dioxide accumulation is a common feature due to longer gas-residence time of mammalian cell culture in large scale bioreactors. A carbon dioxide stripping model can be used to better understand and optimize parameters that are critical to cell culture processes at the manufacturing scale. The prevailing carbon dioxide stripping models in literature depend on mass transfer coefficients and were applicable to cell culture processes with low cell density or at stationary/cell death phase. However, it was reported that gas bubbles are saturated with carbon dioxide before leaving the culture, which makes carbon dioxide stripping no longer depend on a mass transfer coefficient in the new generation cell culture processes characterized by longer exponential growth phase, higher peak viable cell densities, and higher specific production rate. Here, we present a new carbon dioxide stripping model for manufacturing scale bioreactors, which is independent of carbon dioxide mass transfer coefficient, but takes into account the gas-residence time and gas CO 2 saturation time. The model was verified by CHO cell culture processes with different peak viable cell densities (7 to 12 × 10 6  cells mL -1 ) for two products in 5,000-L and 25,000-L bioreactors. The model was also applied to a next generation cell culture process to optimize cell culture conditions and reduce carbon dioxide levels at manufacturing scale. The model provides a useful tool to understand and better control cell culture carbon dioxide

Microfluidic perfusion culture of human induced pluripotent stem cells under fully defined culture conditions.

Science.gov (United States)

Yoshimitsu, Ryosuke; Hattori, Koji; Sugiura, Shinji; Kondo, Yuki; Yamada, Rotaro; Tachikawa, Saoko; Satoh, Taku; Kurisaki, Akira; Ohnuma, Kiyoshi; Asashima, Makoto; Kanamori, Toshiyuki

2014-05-01

Human induced pluripotent stem cells (hiPSCs) are a promising cell source for drug screening. For this application, self-renewal or differentiation of the cells is required, and undefined factors in the culture conditions are not desirable. Microfluidic perfusion culture allows the production of small volume cultures with precisely controlled microenvironments, and is applicable to high-throughput cellular environment screening. Here, we developed a microfluidic perfusion culture system for hiPSCs that uses a microchamber array chip under defined extracellular matrix (ECM) and culture medium conditions. By screening various ECMs we determined that fibronectin and laminin are appropriate for microfluidic devices made out of the most popular material, polydimethylsiloxane (PDMS). We found that the growth rate of hiPSCs under pressure-driven perfusion culture conditions was higher than under static culture conditions in the microchamber array. We applied our new system to self-renewal and differentiation cultures of hiPSCs, and immunocytochemical analysis showed that the state of the hiPSCs was successfully controlled. The effects of three antitumor drugs on hiPSCs were comparable between microchamber array and 96-well plates. We believe that our system will be a platform technology for future large-scale screening of fully defined conditions for differentiation cultures on integrated microfluidic devices. © 2013 Wiley Periodicals, Inc.

Fabrication and perfusion culture of anatomically shaped artificial bone using stereolithography

International Nuclear Information System (INIS)

Du, Dajiang; Asaoka, Teruo; Ushida, Takashi; Furukawa, Katsuko S

2014-01-01

Because patient bone defects are usually varied and complicated in geometry, it would be preferred to fabricate custom-made artificial bone grafts that are anatomically specific to individual patient defects. Using a rabbit femoral segment as a bone reconstruction model, we successfully produced customized ceramic scaffolds by stereolithography, which not only had an anatomically correct external shape according to computed tomography data but also contained an interconnecting internal network of channels designed for perfusion culture. Rabbit bone marrow stromal cells were isolated and cultured with these scaffolds using a novel oscillatory perfusion system that was stereolithographically fabricated to fit well to the unique scaffold shapes. After five days of three-dimensional culture with oscillatory perfusion, the cells attached and proliferated homogenously in the scaffolds. However, control cells inside the scaffolds cultured under static conditions were dead after prolonged in vitro culture. Cellular DNA content and alkaline phosphatase activities were significantly higher in the perfusion group versus the static group. Therefore, anatomically correct artificial bone can be successfully constructed using stereolithography and oscillatory culture technology, and could be useful for bone engraftment and defect repair. (paper)

Streamlined bioreactor-based production of human cartilage tissues.

Science.gov (United States)

Tonnarelli, B; Santoro, R; Adelaide Asnaghi, M; Wendt, D

2016-05-27

Engineered tissue grafts have been manufactured using methods based predominantly on traditional labour-intensive manual benchtop techniques. These methods impart significant regulatory and economic challenges, hindering the successful translation of engineered tissue products to the clinic. Alternatively, bioreactor-based production systems have the potential to overcome such limitations. In this work, we present an innovative manufacturing approach to engineer cartilage tissue within a single bioreactor system, starting from freshly isolated human primary chondrocytes, through the generation of cartilaginous tissue grafts. The limited number of primary chondrocytes that can be isolated from a small clinically-sized cartilage biopsy could be seeded and extensively expanded directly within a 3D scaffold in our perfusion bioreactor (5.4 ± 0.9 doublings in 2 weeks), bypassing conventional 2D expansion in flasks. Chondrocytes expanded in 3D scaffolds better maintained a chondrogenic phenotype than chondrocytes expanded on plastic flasks (collagen type II mRNA, 18-fold; Sox-9, 11-fold). After this "3D expansion" phase, bioreactor culture conditions were changed to subsequently support chondrogenic differentiation for two weeks. Engineered tissues based on 3D-expanded chondrocytes were more cartilaginous than tissues generated from chondrocytes previously expanded in flasks. We then demonstrated that this streamlined bioreactor-based process could be adapted to effectively generate up-scaled cartilage grafts in a size with clinical relevance (50 mm diameter). Streamlined and robust tissue engineering processes, as the one described here, may be key for the future manufacturing of grafts for clinical applications, as they facilitate the establishment of compact and closed bioreactor-based production systems, with minimal automation requirements, lower operating costs, and increased compliance to regulatory guidelines.

Transcriptomics as a tool for assessing the scalability of mammalian cell perfusion systems.

Science.gov (United States)

Jayapal, Karthik P; Goudar, Chetan T

2014-01-01

DNA microarray-based transcriptomics have been used to determine the time course of laboratory and manufacturing-scale perfusion bioreactors in an attempt to characterize cell physiological state at these two bioreactor scales. Given the limited availability of genomic data for baby hamster kidney (BHK) cells, a Chinese hamster ovary (CHO)-based microarray was used following a feasibility assessment of cross-species hybridization. A heat shock experiment was performed using both BHK and CHO cells and resulting DNA microarray data were analyzed using a filtering criteria of perfect match (PM)/single base mismatch (MM) > 1.5 and PM-MM > 50 to exclude probes with low specificity or sensitivity for cross-species hybridizations. For BHK cells, 8910 probe sets (39 %) passed the cutoff criteria, whereas 12,961 probe sets (56 %) passed the cutoff criteria for CHO cells. Yet, the data from BHK cells allowed distinct clustering of heat shock and control samples as well as identification of biologically relevant genes as being differentially expressed, indicating the utility of cross-species hybridization. Subsequently, DNA microarray analysis was performed on time course samples from laboratory- and manufacturing-scale perfusion bioreactors that were operated under the same conditions. A majority of the variability (37 %) was associated with the first principal component (PC-1). Although PC-1 changed monotonically with culture duration, the trends were very similar in both the laboratory and manufacturing-scale bioreactors. Therefore, despite time-related changes to the cell physiological state, transcriptomic fingerprints were similar across the two bioreactor scales at any given instance in culture. Multiple genes were identified with time-course expression profiles that were very highly correlated (> 0.9) with bioprocess variables of interest. Although the current incomplete annotation limits the biological interpretation of these observations, their full potential may be

In Vivo-Like Culture Conditions in a Bioreactor Facilitate Improved Tissue Quality in Corneal Storage.

Science.gov (United States)

Schmid, Richard; Tarau, Ioana-Sandra; Rossi, Angela; Leonhardt, Stefan; Schwarz, Thomas; Schuerlein, Sebastian; Lotz, Christian; Hansmann, Jan

2018-01-01

The cornea is the most-transplanted tissue worldwide. However, the availability and quality of grafts are limited due to the current methods of corneal storage. In this study, a dynamic bioreactor system is employed to enable the control of intraocular pressure and the culture at the air-liquid interface. Thereby, in vivo-like storage conditions are achieved. Different media combinations for endothelium and epithelium are tested in standard and dynamic conditions to enhance the viability of the tissue. In contrast to culture conditions used in eye banks, the combination of the bioreactor and biochrom medium 1 allows to preserve the corneal endothelium and the epithelium. Assessment of transparency, swelling, and the trans-epithelial-electrical-resistance (TEER) strengthens the impact of the in vivo-like tissue culture. For example, compared to corneas stored under static conditions, significantly lower optical densities and significantly higher TEER values were measured (p-value <0.05). Furthermore, healing of epithelial defects is enabled in the bioreactor, characterized by re-epithelialization and initiated stromal regeneration. Based on the obtained results, an easy-to-use 3D-printed bioreactor composed of only two parts was derived to translate the technology from the laboratory to the eye banks. This optimized bioreactor facilitates noninvasive microscopic monitoring. The improved storage conditions ameliorate the quality of corneal grafts and the storage time in the eye banks to increase availability and reduce re-grafting. © 2017 The Authors. Biotechnology Journal Published by Wiley-VCH Verlag GmbH & Co. KGaA.

Production of oncolytic adenovirus and human mesenchymal stem cells in a single-use, Vertical-Wheel bioreactor system: Impact of bioreactor design on performance of microcarrier-based cell culture processes.

Science.gov (United States)

Sousa, Marcos F Q; Silva, Marta M; Giroux, Daniel; Hashimura, Yas; Wesselschmidt, Robin; Lee, Brian; Roldão, António; Carrondo, Manuel J T; Alves, Paula M; Serra, Margarida

2015-01-01

Anchorage-dependent cell cultures are used for the production of viruses, viral vectors, and vaccines, as well as for various cell therapies and tissue engineering applications. Most of these applications currently rely on planar technologies for the generation of biological products. However, as new cell therapy product candidates move from clinical trials towards potential commercialization, planar platforms have proven to be inadequate to meet large-scale manufacturing demand. Therefore, a new scalable platform for culturing anchorage-dependent cells at high cell volumetric concentrations is urgently needed. One promising solution is to grow cells on microcarriers suspended in single-use bioreactors. Toward this goal, a novel bioreactor system utilizing an innovative Vertical-Wheel™ technology was evaluated for its potential to support scalable cell culture process development. Two anchorage-dependent human cell types were used: human lung carcinoma cells (A549 cell line) and human bone marrow-derived mesenchymal stem cells (hMSC). Key hydrodynamic parameters such as power input, mixing time, Kolmogorov length scale, and shear stress were estimated. The performance of Vertical-Wheel bioreactors (PBS-VW) was then evaluated for A549 cell growth and oncolytic adenovirus type 5 production as well as for hMSC expansion. Regarding the first cell model, higher cell growth and number of infectious viruses per cell were achieved when compared with stirred tank (ST) bioreactors. For the hMSC model, although higher percentages of proliferative cells could be reached in the PBS-VW compared with ST bioreactors, no significant differences in the cell volumetric concentration and expansion factor were observed. Noteworthy, the hMSC population generated in the PBS-VW showed a significantly lower percentage of apoptotic cells as well as reduced levels of HLA-DR positive cells. Overall, these results showed that process transfer from ST bioreactor to PBS-VW, and scale-up was

Expansion of Bone Marrow Mesenchymal Stromal Cells in Perfused 3D Ceramic Scaffolds Enhances In Vivo Bone Formation.

Science.gov (United States)

Hoch, Allison I; Duhr, Ralph; Di Maggio, Nunzia; Mehrkens, Arne; Jakob, Marcel; Wendt, David

2017-12-01

Bone marrow-derived mesenchymal stromal cells (BMSC), when expanded directly within 3D ceramic scaffolds in perfusion bioreactors, more reproducibly form bone when implanted in vivo as compared to conventional expansion on 2D polystyrene dishes/flasks. Since the bioreactor-based expansion on 3D ceramic scaffolds encompasses multiple aspects that are inherently different from expansion on 2D polystyrene, we aimed to decouple the effects of specific parameters among these two model systems. We assessed the effects of the: 1) 3D scaffold vs. 2D surface; 2) ceramic vs. polystyrene materials; and 3) BMSC niche established within the ceramic pores during in vitro culture, on subsequent in vivo bone formation. While BMSC expanded on 3D polystyrene scaffolds in the bioreactor could maintain their in vivo osteogenic potential, results were similar as BMSC expanded in monolayer on 2D polystyrene, suggesting little influence of the scaffold 3D environment. Bone formation was most reproducible when BMSC are expanded on 3D ceramic, highlighting the influence of the ceramic substrate. The presence of a pre-formed niche within the scaffold pores had negligible effects on the in vivo bone formation. The results of this study allow a greater understanding of the parameters required for perfusion bioreactor-based manufacturing of osteogenic grafts for clinical applications. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Comparing effects of perfusion and hydrostatic pressure on gene profiles of human chondrocyte.

Science.gov (United States)

Zhu, Ge; Mayer-Wagner, Susanne; Schröder, Christian; Woiczinski, Matthias; Blum, Helmut; Lavagi, Ilaria; Krebs, Stefan; Redeker, Julia I; Hölzer, Andreas; Jansson, Volkmar; Betz, Oliver; Müller, Peter E

2015-09-20

Hydrostatic pressure and perfusion have been shown to regulate the chondrogenic potential of articular chondrocytes. In order to compare the effects of hydrostatic pressure plus perfusion (HPP) and perfusion (P) we investigated the complete gene expression profiles of human chondrocytes under HPP and P. A simplified bioreactor was constructed to apply loading (0.1 MPa for 2 h) and perfusion (2 ml) through the same piping by pressurizing the medium directly. High-density monolayer cultures of human chondrocytes were exposed to HPP or P for 4 days. Controls (C) were maintained in static cultures. Gene expression was evaluated by sequencing (RNAseq) and quantitative real-time PCR analysis. Both treatments changed gene expression levels of human chondrocytes significantly. Specifically, HPP and P increased COL2A1 expression and decreased COL1A1 and MMP-13 expression. Despite of these similarities, RNAseq revealed a list of cartilage genes including ACAN, ITGA10 and TNC, which were differentially expressed by HPP and P. Of these candidates, adhesion related molecules were found to be upregulated in HPP. Both HPP and P treatment had beneficial effects on chondrocyte differentiation and decreased catabolic enzyme expression. The study provides new insight into how hydrostatic pressure and perfusion enhance cartilage differentiation and inhibit catabolic effects. Copyright © 2015 Elsevier B.V. All rights reserved.

Bioreactor technology for herbal plants

International Nuclear Information System (INIS)

Sobri Hussein; Rusli Ibrahim; Abdul Rahim Harun; Azhar Mohamad; Hawa Abdul Aziz; Wan Nazirah Wan Ali

2010-01-01

Plants have been an important source of medicine for thousands of years and herbs are hot currency in the world today. During the last decade, popularity of alternative medicine increased significantly worldwide with noticeable trend. This in turn accelerated the global trade of herbal raw materials and herbal products and created greater scope for Asian countries that possess the major supply of herbal raw materials within their highly diversified tropical rain forest. As such, advanced bioreactor culture system possesses a great potential for large scale production than the traditional tissue culture system. Bioreactor cultures have many advantages over conventional cultures. Plant cells in bioreactors can grow fast and vigorously in shorter period as the culture conditions in bioreactor such as temperature, pH, concentrations of dissolved oxygen, carbon dioxide and nutrients can be optimised by on-line manipulation. Nutrient uptake can also be enhanced by continuous medium circulation, which ultimately increased cell proliferation rate. Consequently, production period and cost are substantially reduced, product quality is controlled and standardized as well as free of pesticide contamination and production of raw material can be conducted all year round. Taking all these into consideration, current research efforts were focused on varying several parameters such as inoculation density, air flow, medium formulation, PGRs etc. for increased production of cell and organ cultures of high market demand herbal and medicinal plants, particularly Eurycoma longifolia, Panax ginseng and Labisia pumila. At present, the production of cell and organ culture of these medicinal plants have also been applied in airlift bioreactor with different working volumes. It is hope that the investment of research efforts into this advanced bioreactor technology will open up a bright future for the modernization of agriculture and commercialisation of natural product. (author)

Perfusion directed 3D mineral formation within cell-laden hydrogels.

Science.gov (United States)

Sawyer, Stephen William; Shridhar, Shivkumar Vishnempet; Zhang, Kairui; Albrecht, Lucas; Filip, Alex; Horton, Jason; Soman, Pranav

2018-06-08

Despite the promise of stem cell engineering and the new advances in bioprinting technologies, one of the major challenges in the manufacturing of large scale bone tissue scaffolds is the inability to perfuse nutrients throughout thick constructs. Here, we report a scalable method to create thick, perfusable bone constructs using a combination of cell-laden hydrogels and a 3D printed sacrificial polymer. Osteoblast-like Saos-2 cells were encapsulated within a gelatin methacrylate (GelMA) hydrogel and 3D printed polyvinyl alcohol (PVA) pipes were used to create perfusable channels. A custom-built bioreactor was used to perfuse osteogenic media directly through the channels in order to induce mineral deposition which was subsequently quantified via microCT. Histological staining was used to verify mineral deposition around the perfused channels, while COMSOL modeling was used to simulate oxygen diffusion between adjacent channels. This information was used to design a scaled-up construct containing a 3D array of perfusable channels within cell-laden GelMA. Progressive matrix mineralization was observed by cells surrounding perfused channels as opposed to random mineral deposition in static constructs. MicroCT confirmed that there was a direct relationship between channel mineralization within perfused constructs and time within the bioreactor. Furthermore, the scalable method presented in this work serves as a model on how large-scale bone tissue replacement constructs could be made using commonly available 3D printers, sacrificial materials, and hydrogels. © 2018 IOP Publishing Ltd.

MELiSSA third compartment: Nitrosomonas europaea and Nitrobacter winogradskyi axenic cultures in bioreactors

Science.gov (United States)

Cruvellier, Nelly; Lasseur, Christophe; Poughon, Laurent; Creuly, Catherine; Dussap, Gilles

Nitrogen is a key element for the life and its balance on Earth is regulated by the nitrogen cycle. This loop includes several steps among which nitrification that permits the transformation of the ammonium into nitrate. The MELiSSA loop is an artificial ecosystem designed for life support systems (LSS). It is based on the carbon and nitrogen cycles and the recycling of the non-edible part of the higher plants and the waste produced by the crew. In this order, all the wastes are collected in the first compartment to degrade them into organic acids and CO2. These compounds are joining the second compartment which is a photoheterotrophic compartment where at the outlet an organic-free medium containing ammonium is produced. This solution will be the substrate of the third compartment where nitrification is done. This compartment has to oxidize the ammonium into nitrate, and this biological reaction needs two steps. In the MELiSSA loop, the nitrification is carried out by two bacteria: Nitrosomonas europaea ATCC® 19718™ which is oxidizing ammonia into nitrite and Nitrobacter winogradskyi ATCC® 25391™ which is producing nitrate from nitrite in the third compartment. These two bacteria are growing in axenic conditions on a fixed bed bioreactor filled with Biostyr® beads. The nitrogen compounds are controlled by Ionic Chromatography and colorimetric titration for each sample. The work presented here deals with the culture of both bacteria in pure cultures and mixed cultures in stirred and aerated bioreactors of different volumes. The first aim of our work is the characterization of the bacteria growth in bioreactors and in the nitrifying fixed-bed column. The experimental results confirm that the growth is slow; the maximal growth rate in suspended cultures is 0.054h-1 for Nitrosomonas europaea and 0.022h-1 for Nitrobacter winogradskyi. Mixed cultures are difficult to control and operate but one could be done for more than 500 hours. The characterization of the

Feasibility of direct oxygenation of primary-cultured rat hepatocytes using polyethylene glycol-decorated liposome-encapsulated hemoglobin (LEH).

Science.gov (United States)

Naruto, Hirosuke; Huang, Hongyun; Nishikawa, Masaki; Kojima, Nobuhiko; Mizuno, Atsushi; Ohta, Katsuji; Sakai, Yasuyuki

2007-10-01

We tested the short-term efficacy of liposome-encapsulated hemoglobin (LEH) in cultured rat hepatocytes. Supplementation with LEH (20% of the hemoglobin concentration of blood) did not lower albumin production in static culture, and completely reversed the cell death and deterioration in albumin production caused by an oxygen shortage in 2D flat-plate perfusion bioreactors.

Hepatic Differentiation of Human Induced Pluripotent Stem Cells in a Perfused 3D Porous Polymer Scaffold for Liver Tissue Engineering

DEFF Research Database (Denmark)

Hemmingsen, Mette; Muhammad, Haseena Bashir; Mohanty, Soumyaranjan

A huge shortage of liver organs for transplantation has motivated the research field of tissue engineering to develop bioartificial liver tissue and even a whole liver. The goal of NanoBio4Trans is to create a vascularized bioartificial liver tissue, initially as a liver-support system. Due...... to limitations of primary hepatocytes regarding availability and maintenance of functionality, stem cells and especially human induced pluripotent stem cells (hIPS cells) are an attractive cell source for liver tissue engineering. The aim of this part of NanoBio4Trans is to optimize culture and hepatic...... differentiation of hIPS-derived definitive endoderm (DE) cells in a 3D porous polymer scaffold built-in a perfusable bioreactor. The use of a microfluidic bioreactor array enables the culture of 16 independent tissues in one experimental run and thereby an optimization study to be performed....

Comparison of spectroscopy technologies for improved monitoring of cell culture processes in miniature bioreactors

DEFF Research Database (Denmark)

Rowland-Jones, Ruth C.; van der Berg, Franciscus Winfried J; Racher, Andrew J.

2017-01-01

Cell culture process development requires the screening of large numbers of cell lines and process conditions. The development of miniature bioreactor systems has increased the throughput of such studies; however, there are limitations with their use. One important constraint is the limited numbe...

Scaling down of a clinical three-dimensional perfusion multicompartment hollow fiber liver bioreactor developed for extracorporeal liver support to an analytical scale device useful for hepatic pharmacological in vitro studies.

Science.gov (United States)

Zeilinger, Katrin; Schreiter, Thomas; Darnell, Malin; Söderdahl, Therese; Lübberstedt, Marc; Dillner, Birgitta; Knobeloch, Daniel; Nüssler, Andreas K; Gerlach, Jörg C; Andersson, Tommy B

2011-05-01

Within the scope of developing an in vitro culture model for pharmacological research on human liver functions, a three-dimensional multicompartment hollow fiber bioreactor proven to function as a clinical extracorporeal liver support system was scaled down in two steps from 800 mL to 8 mL and 2 mL bioreactors. Primary human liver cells cultured over 14 days in 800, 8, or 2 mL bioreactors exhibited comparable time-course profiles for most of the metabolic parameters in the different bioreactor size variants. Major drug-metabolizing cytochrome P450 activities analyzed in the 2 mL bioreactor were preserved over up to 23 days. Immunohistochemical studies revealed tissue-like structures of parenchymal and nonparenchymal cells in the miniaturized bioreactor, indicating physiological reorganization of the cells. Moreover, the canalicular transporters multidrug-resistance-associated protein 2, multidrug-resistance protein 1 (P-glycoprotein), and breast cancer resistance protein showed a similar distribution pattern to that found in human liver tissue. In conclusion, the down-scaled multicompartment hollow fiber technology allows stable maintenance of primary human liver cells and provides an innovative tool for pharmacological and kinetic studies of hepatic functions with small cell numbers.

The effect of hydroxylamine on the activity and aggregate structure of autotrophic nitrifying bioreactor cultures

DEFF Research Database (Denmark)

Harper, W.F.; Terada, Akihiko; Poly, F.

2009-01-01

Addition of hydroxylamine (NH2OH) to autotrophic biomass in nitrifying bioreactors affected the activity, physical structure, and microbial ecology of nitrifying aggregates. When NH2OH is added to nitrifying cultures in 6-h batch experiments, the initial NH3-N uptake rates were physiologically...... accelerated by a factor of 1.4-13. NH2OH addition caused a 20-40% decrease in the median aggregate size, broadened the shape of the aggregate size distribution by up to 230%, and caused some of the microcolonies to appear slightly more dispersed. Longer term NH2OH addition in fed batch bioreactors decreased...

Production of poly-beta-hydroxybutyric acid by microorganisms accumulated from river water using a two-stage perfusion culture system.

Science.gov (United States)

Morimoto, T; Tashiro, F; Nagashima, H; Nishizawa, K; Nagata, F; Yokogawa, Y; Suzuki, T

2000-01-01

The perfusion culture system using a shaken ceramic membrane flask (SCMF) was employed to accumulate microorganisms separated from river water and to produce poly-beta-hydroxybutyric acid (PHB). Using a two-step culture method with a single SCMF, river microorganisms were cultured by separately feeding four representative carbon sources, n-propanol, lactic acid, methanol, and formic acid. After 140 h culture, the cell concentration and PHB content respectively reached 43 g/l and 35% when a propanol medium was fed. Using a two-stage perfusion culture with twin SCMFs, the seed cell mass was increased in the first SCMF and then supplied to the second flask for PHB production. As a consequence, the cellular PHB content rose to 51% in the second SCMF, while the cell concentration gradually increased to 25 g/l after 175 h perfusion culture. These results demonstrated the utility of the two-stage perfusion culture system for developing a cheap means of producing PHB coincident with wastewater treatment.

A 3D printed microfluidic perfusion device for multicellular spheroid cultures.

Science.gov (United States)

Ong, Louis Jun Ye; Islam, Anik; DasGupta, Ramanuj; Iyer, Narayanan Gopalakkrishna; Leo, Hwa Liang; Toh, Yi-Chin

2017-09-11

The advent of 3D printing technologies promises to make microfluidic organ-on-chip technologies more accessible for the biological research community. To date, hydrogel-encapsulated cells have been successfully incorporated into 3D printed microfluidic devices. However, there is currently no 3D printed microfluidic device that can support multicellular spheroid culture, which facilitates extensive cell-cell contacts important for recapitulating many multicellular functional biological structures. Here, we report a first instance of fabricating a 3D printed microfluidic cell culture device capable of directly immobilizing and maintaining the viability and functionality of 3D multicellular spheroids. We evaluated the feasibility of two common 3D printing technologies i.e. stereolithography (SLA) and PolyJet printing, and found that SLA could prototype a device comprising of cell immobilizing micro-structures that were housed within a microfluidic network with higher fidelity. We have also implemented a pump-free perfusion system, relying on gravity-driven flow to perform medium perfusion in order to reduce the complexity and footprint of the device setup, thereby improving its adaptability into a standard biological laboratory. Finally, we demonstrated the biological performance of the 3D printed device by performing pump-free perfusion cultures of patient-derived parental and metastatic oral squamous cell carcinoma tumor and liver cell (HepG2) spheroids with good cell viability and functionality. This paper presents a proof-of-concept in simplifying and integrating the prototyping and operation of a microfluidic spheroid culture device, which will facilitate its applications in various drug efficacy, metabolism and toxicity studies.

Improved Performance in Mammalian Cell Perfusion Cultures by Growth Inhibition.

Science.gov (United States)

Wolf, Moritz K F; Closet, Aurélie; Bzowska, Monika; Bielser, Jean-Marc; Souquet, Jonathan; Broly, Hervé; Morbidelli, Massimo

2018-05-21

Mammalian cell perfusion cultures represent a promising alternative to the current fed-batch technology for the production of various biopharmaceuticals. Long-term operation at a fixed viable cell density (VCD) requires a viable culture and a constant removal of excessive cells. Product loss in the cell removing bleed stream deteriorates the process yield. In this study, the authors investigate the use of chemical and environmental growth inhibition on culture performance by either adding valeric acid (VA) to the production media or by reducing the culture temperature (33.0 °C) with respect to control conditions (36.5 °C, no VA). Low temperature significantly reduces cellular growth, thus, resulting in lower bleed rates accompanied by a reduced product loss of 11% compared to 26% under control conditions. Additionally, the cell specific productivity of the target protein improves and maintained stable leading to media savings per mass of product. VA shows initially an inhibitory effect on cellular growth. However, cells seemed to adapt to the presence of the inhibitor resulting in a recovery of the cellular growth. Cell cycle and Western blot analyses support the observed results. This work underlines the role of temperature as a key operating variable for the optimization of perfusion cultures. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Production Pattern of Ajmalicine in Catharanthus roseus (L. G. Don. Cell Aggregates Culture in the Airlift Bioreactor

Directory of Open Access Journals (Sweden)

RIZKITA RACHMI ESYANTI

2006-12-01

Full Text Available A research has been conducted to optimize the rate of aeration and initial weight of cell aggregates in the production of ajmalicine in Catharanthus roseus cell culture in airlift bioreactor. Catharanthus roseus culture were grown in Zenk medium with the addition of 2.50 x 10-6 M naphthalene acetic acid (NAA and 10-5 M benzyl amino purine (BAP. Cell aggregates were sub-cultured two times before transferring 20 and 30 g/fw of cell aggregates into bioreactor, respectively, and aerated with the rate of 0.25 l min-1 and 0.34 l min-1, respectively. The pattern of ajmalicine production in bioreactor were observed in every three days within 24 days. Qualitative and quantitative analysis were conducted using HPLC connected to Cromatopac CL-7A Plus. The results showed that the cell aggregates and medium contain ajmalicine. The highest concentration was obtained in combination of 30 g/fw and 0.34 l min-1 aeration compare to 20 g/fw - 0.25 l min-1, 20 g/fw - 0.34 l min-1, as well as 30 g/fw – 0.25 l min-1. The highest ajmalicine content in cell aggregates was obtained on the 12 days (79.23 µg g-1 whilst in medium was obtained in the 18th days (981.15 µg l-1.

A Novel bioreactor with mechanical stimulation for skeletal tissue engineering

Directory of Open Access Journals (Sweden)

M. Petrović

2009-01-01

Full Text Available The provision of mechanical stimulation is believed to be necessary for the functional assembly of skeletal tissues, which are normally exposed to a variety of biomechanical signals in vivo. In this paper, we present a development and validation of a novel bioreactor aimed for skeletal tissue engineering that provides dynamic compression and perfusion of cultivated tissues. Dynamic compression can be applied at frequencies up to 67.5 Hz and displacements down to 5 m thus suitable for the simulation of physiological conditions in a native cartilage tissue (0.1-1 Hz, 5-10 % strain. The bioreactor also includes a load sensor that was calibrated so to measure average loads imposed on tissue samples. Regimes of the mechanical stimulation and acquisition of load sensor outputs are directed by an automatic control system using applications developed within the LabView platform. In addition, perfusion of tissue samples at physiological velocities (10–100 m/s provides efficient mass transfer, as well as the possibilities to expose the cells to hydrodynamic shear and simulate the conditions in a native bone tissue. Thus, the novel bioreactor is suited for studies of the effects of different biomechanical signals on in vitro regeneration of skeletal tissues, as well as for the studies of newly formulated biomaterials and cell biomaterial interactions under in vivo-like settings.

Microfluidically supported biochip design for culture of endothelial cell layers with improved perfusion conditions.

Science.gov (United States)

Raasch, Martin; Rennert, Knut; Jahn, Tobias; Peters, Sven; Henkel, Thomas; Huber, Otmar; Schulz, Ingo; Becker, Holger; Lorkowski, Stefan; Funke, Harald; Mosig, Alexander

2015-03-02

Hemodynamic forces generated by the blood flow are of central importance for the function of endothelial cells (ECs), which form a biologically active cellular monolayer in blood vessels and serve as a selective barrier for macromolecular permeability. Mechanical stimulation of the endothelial monolayer induces morphological remodeling in its cytoskeleton. For in vitro studies on EC biology culture devices are desirable that simulate conditions of flow in blood vessels and allow flow-based adhesion/permeability assays under optimal perfusion conditions. With this aim we designed a biochip comprising a perfusable membrane that serves as cell culture platform multi-organ-tissue-flow (MOTiF biochip). This biochip allows an effective supply with nutrition medium, discharge of catabolic cell metabolites and defined application of shear stress to ECs under laminar flow conditions. To characterize EC layers cultured in the MOTiF biochip we investigated cell viability, expression of EC marker proteins and cell adhesion molecules of ECs dynamically cultured under low and high shear stress, and compared them with an endothelial culture in established two-dimensionally perfused flow chambers and under static conditions. We show that ECs cultured in the MOTiF biochip form a tight EC monolayer with increased cellular density, enhanced cell layer thickness, presumably as the result of a rapid and effective adaption to shear stress by remodeling of the cytoskeleton. Moreover, endothelial layers in the MOTiF biochip express higher amounts of EC marker proteins von-Willebrand-factor and PECAM-1. EC layers were highly responsive to stimulation with TNFα as detected at the level of ICAM-1, VCAM-1 and E-selectin expression and modulation of endothelial permeability in response to TNFα/IFNγ treatment under flow conditions. Compared to static and two-dimensionally perfused cell culture condition we consider MOTiF biochips as a valuable tool for studying EC biology in vitro under

Functional study on two artificial liver bioreactors with collagen gel

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XU Bing

2014-10-01

Full Text Available ObjectiveTo improve the hollow fiber bioreactor of artificial liver. MethodsRat hepatocytes mixed with collagen solution were injected into the external cavity of a hollow fiber reactor to construct a bioreactor of hepatocytes suspended in collagen gel (group Ⅰ. Other rat hepatocytes suspended in solution were injected into the external cavity of a hollow fiber reactor with a layer of collagen on the wall of the external cavity to construct a bioreactor of collagen layer and hepatocytes (group Ⅱ. For each group, the culture solution circulated through the internal cavity of the hollow fiber bioreactor; the bioreactor was put in a culture box for 9 d, and the culture solution in the internal cavity was exchanged for new one every 24 h; the concentrations of albumin (Alb, urea, and lactate dehydrogenase (LDH in the culture solution samples were measured to examine the hepatocyte function of the bioreactor. Statistical analysis was performed using SPSS 130. Continuous data were expressed as mean±SD, and comparison between groups was made by paired t test. ResultsFor groups Ⅰ and Ⅱ, Alb levels reached peak values on day 3 of culture (1.41±0.08 g/L and 0.65±0.05 g/L; from day 3 to 9, group I had a significantly higher Alb level than group Ⅱ (t＞7.572, P＜0.01. For groups Ⅰ and Ⅱ, urea levels reached peak values on days 3 and 5 of culture (1.73±0.14 mmol/L and 1.56±0.18 mmol/L; from days 5 to 9, group I had a significantly higher urea level than group Ⅱ (t＞8.418, P＜0.01. For groups Ⅰ and Ⅱ, LDH levels reached peak values on day 9 of culture (32.03±9.13 U/L and 70.17±25.28 U/L; from days 1 to 9, group I had a significantly lower LDH level than group Ⅱ(t＞5.633, P＜0.01. Therefore, the bioreactor of hepatocytes suspended in collagen gel (group Ⅰ showed a better hepatocyte function and less hepatic enzyme leakage compared with the bioreactor of collagen layer and hepatocytes (group Ⅱ. Conclusion

Genetic Algorithmic Optimization of PHB Production by a Mixed Culture in an Optimally Dispersed Fed-batch Bioreactor

Directory of Open Access Journals (Sweden)

Pratap R. Patnaik

2009-10-01

Full Text Available Poly-β-hydroxybutyrate (PHB is an energy-storage polymer whose properties are similar to those of chemical polymers such as polyethylene and polypropylene. Moreover, PHB is biodegradable, absorbed by human tissues and less energy-consuming than synthetic polymers. Although Ralstonia eutropha is widely used to synthesize PHB, it is inefficient in utilizing glucose and similar sugars. Therefore a co-culture of R. eutropha and Lactobacillus delbrueckii is preferred since the latter can convert glucose to lactate, which R. eutropha can metabolize easily. Tohyama et al. [24] maximized PHB production in a well-mixed fed-batch bioreactor with glucose and (NH42SO4 as the primary substrates. Since production-scale bioreactors often deviate from ideal laboratory-scale reactors, a large bioreactor was simulated by means of a dispersion model with the kinetics determined by Tohyama et al. [24] and dispersion set at an optimum Peclet number of 20 [32]. The time-dependent feed rates of the two substrates were determined through a genetic algorithm (GA to maximize PHB production. This bioreactor produced 22.2% more PHB per liter and 12.8% more cell mass than achieved by Tohyama et al. [24]. These results, and similar observations with other fermentations, indicate the feasibility of enhancing the efficiency of large nonideal bioreactors through GA optimizations.

A novel bioreactor to simulate urinary bladder mechanical properties and compliance for bladder functional tissue engineering.

Science.gov (United States)

Wei, Xin; Li, Dao-bing; Xu, Feng; Wang, Yan; Zhu, Yu-chun; Li, Hong; Wang, Kun-jie

2011-02-01

Bioreactors are pivotal tools for generating mechanical stimulation in functional tissue engineering study. This study aimed to create a bioreactor that can simulate urinary bladder mechanical properties, and to investigate the effects of a mechanically stimulated culture on urothelial cells and bladder smooth muscle cells. We designed a bioreactor to simulate the mechanical properties of bladder. A pressure-record system was used to evaluate the mechanical properties of the bioreactor by measuring the pressure in culture chambers. To test the biocompatibility of the bioreactor, viabilities of urothelial cells and smooth muscle cells cultured in the bioreactor under static and mechanically changed conditions were measured after 7-day culture. To evaluate the effect of mechanical stimulations on the vital cells, urethral cells and smooth muscle cells were cultured in the simulated mechanical conditions. After that, the viability and the distribution pattern of the cells were observed and compared with cells cultured in non-mechanical stimulated condition. The bioreactor system successfully generated waveforms similar to the intended programmed model while maintaining a cell-seeded elastic membrane between the chambers. There were no differences between viabilities of urothelial cells ((91.90 ± 1.22)% vs. (93.14 ± 1.78)%, P > 0.05) and bladder smooth muscle cells ((93.41 ± 1.49)% vs. (92.61 ± 1.34)%, P > 0.05). The viability of cells and tissue structure observation after cultured in simulated condition showed that mechanical stimulation was the only factor affected cells in the bioreactor and improved the arrangement of cells on silastic membrane. This bioreactor can effectively simulate the physiological and mechanical properties of the bladder. Mechanical stimulation is the only factor that affected the viability of cells cultured in the bioreactor. The bioreactor can change the growth behavior of urothelial cells and bladder smooth muscle cells, resulting in

Schisandra lignans production regulated by different bioreactor type.

Science.gov (United States)

Szopa, Agnieszka; Kokotkiewicz, Adam; Luczkiewicz, Maria; Ekiert, Halina

2017-04-10

Schisandra chinensis (Chinese magnolia vine) is a rich source of therapeutically relevant dibenzocyclooctadiene lignans with anticancer, immunostimulant and hepatoprotective activities. In this work, shoot cultures of S. chinensis were grown in different types of bioreactors with the aim to select a system suitable for the large scale in vitro production of schisandra lignans. The cultures were maintained in Murashige-Skoog (MS) medium supplemented with 3mg/l 6-benzylaminopurine (BA) and 1mg/l 1-naphthaleneacetic acid (NAA). Five bioreactors differing with respect to cultivation mode were tested: two liquid-phase systems (baloon-type bioreactor and bubble-column bioreactor with biomass immobilization), the gas-phase spray bioreactor and two commercially available temporary immersion systems: RITA ® and Plantform. The experiments were run for 30 and 60 days in batch mode. The harvested shoots were evaluated for growth and lignan content determined by LC-DAD and LC-DAD-ESI-MS. Of the tested bioreactors, temporary immersion systems provided the best results with respect to biomass production and lignan accumulation: RITA ® bioreactor yielded 17.86g/l (dry weight) during 60 day growth period whereas shoots grown for 30 days in Plantform bioreactor contained the highest amount of lignans (546.98mg/100g dry weight), with schisandrin, deoxyschisandrin and gomisin A as the major constituents (118.59, 77.66 and 67.86mg/100g dry weight, respectively). Copyright © 2017 Elsevier B.V. All rights reserved.

Fed-batch and perfusion culture processes: economic, environmental, and operational feasibility under uncertainty.

Science.gov (United States)

Pollock, James; Ho, Sa V; Farid, Suzanne S

2013-01-01

This article evaluates the current and future potential of batch and continuous cell culture technologies via a case study based on the commercial manufacture of monoclonal antibodies. The case study compares fed-batch culture to two perfusion technologies: spin-filter perfusion and an emerging perfusion technology utilizing alternating tangential flow (ATF) perfusion. The operational, economic, and environmental feasibility of whole bioprocesses based on these systems was evaluated using a prototype dynamic decision-support tool built at UCL encompassing process economics, discrete-event simulation and uncertainty analysis, and combined with a multi-attribute decision-making technique so as to enable a holistic assessment. The strategies were compared across a range of scales and titres so as to visualize how their ranking changes in different industry scenarios. The deterministic analysis indicated that the ATF perfusion strategy has the potential to offer cost of goods savings of 20% when compared to conventional fed-batch manufacturing processes when a fivefold increase in maximum viable cell densities was assumed. Savings were also seen when the ATF cell density dropped to a threefold increase over the fed-batch strategy for most combinations of titres and production scales. In contrast, the fed-batch strategy performed better in terms of environmental sustainability with a lower water and consumable usage profile. The impact of uncertainty and failure rates on the feasibility of the strategies was explored using Monte Carlo simulation. The risk analysis results demonstrated the enhanced robustness of the fed-batch process but also highlighted that the ATF process was still the most cost-effective option even under uncertainty. The multi-attribute decision-making analysis provided insight into the limited use of spin-filter perfusion strategies in industry. The resulting sensitivity spider plots enabled identification of the critical ratio of weightings of

Dynamic Compression Promotes the Matrix Synthesis of Nucleus Pulposus Cells Through Up-Regulating N-CDH Expression in a Perfusion Bioreactor Culture.

Science.gov (United States)

Xu, Yichun; Yao, Hui; Li, Pei; Xu, Wenbin; Zhang, Junbin; Lv, Lulu; Teng, Haijun; Guo, Zhiliang; Zhao, Huiqing; Hou, Gang

2018-01-01

An adequate matrix production of nucleus pulposus (NP) cells is an important tissue engineering-based strategy to regenerate degenerative discs. Here, we mainly aimed to investigate the effects and mechanism of mechanical compression (i.e., static compression vs. dynamic compression) on the matrix synthesis of three-dimensional (3D) cultured NP cells in vitro. Rat NP cells seeded on small intestinal submucosa (SIS) cryogel scaffolds were cultured in the chambers of a self-developed, mechanically active bioreactor for 10 days. Meanwhile, the NP cells were subjected to compression (static compression or dynamic compression at a 10% scaffold deformation) for 6 hours once per day. Unloaded NP cells were used as controls. The cellular phenotype and matrix biosynthesis of NP cells were investigated by real-time PCR and Western blotting assays. Lentivirus-mediated N-cadherin (N-CDH) knockdown and an inhibitor, LY294002, were used to further investigate the role of N-CDH and the PI3K/Akt pathway in this process. Dynamic compression better maintained the expression of cell-specific markers (keratin-19, FOXF1 and PAX1) and matrix macromolecules (aggrecan and collagen II), as well as N-CDH expression and the activity of the PI3K/Akt pathway, in the 3D-cultured NP cells compared with those expression levels and activity in the cells grown under static compression. Further analysis showed that the N-CDH knockdown significantly down-regulated the expression of NP cell-specific markers and matrix macromolecules and inhibited the activation of the PI3K/Akt pathway under dynamic compression. However, inhibition of the PI3K/Akt pathway had no effects on N-CDH expression but down-regulated the expression of NP cell-specific markers and matrix macromolecules under dynamic compression. Dynamic compression increases the matrix synthesis of 3D-cultured NP cells compared with that of the cells under static compression, and the N-CDH-PI3K/Akt pathway is involved in this regulatory process

Mass culture of mountain Ginseng roots using rare earth elements in bioreactor cultures

Energy Technology Data Exchange (ETDEWEB)

Hwang, Sung Jin; Kim, Chang Hyun; Kim, Ha Lim [Chonnam National University, Gwangju (Korea, Republic of)

2010-01-15

An adventitious roots of mountain ginseng (Panax ginseng C. A. Meyer) was used in this experiments. Various concentration of lanthanide were tested to find out optimal conditions for biomass and ginsenoside contents in mountain ginseng roots. The MS basal medium with 100 {mu}g/L lanthanide created the most optimum condition for growth of adventitious roots of mountain ginseng. Batch culture with 100 {mu}g/L lanthanide and 0.5 g (F.W) inoculation volume produced maximum final biomass of 1.89 g(F.W/flask) within 4 weeks. However, lanthanide was not effect the ginsenoside contents in adventitious roots of mountain ginseng. In bioreactors, 3.23 g F.W./L of biomass were obtained when 100 {mu}g/L lanthanide were added to the MS basal medium at 26 .deg. C

Bioreactors to influence stem cell fate: augmentation of mesenchymal stem cell signaling pathways via dynamic culture systems.

Science.gov (United States)

Yeatts, Andrew B; Choquette, Daniel T; Fisher, John P

2013-02-01

Mesenchymal stem cells (MSCs) are a promising cell source for bone and cartilage tissue engineering as they can be easily isolated from the body and differentiated into osteoblasts and chondrocytes. A cell based tissue engineering strategy using MSCs often involves the culture of these cells on three-dimensional scaffolds; however the size of these scaffolds and the cell population they can support can be restricted in traditional static culture. Thus dynamic culture in bioreactor systems provides a promising means to culture and differentiate MSCs in vitro. This review seeks to characterize key MSC differentiation signaling pathways and provides evidence as to how dynamic culture is augmenting these pathways. Following an overview of dynamic culture systems, discussion will be provided on how these systems can effectively modify and maintain important culture parameters including oxygen content and shear stress. Literature is reviewed for both a highlight of key signaling pathways and evidence for regulation of these signaling pathways via dynamic culture systems. The ability to understand how these culture systems are affecting MSC signaling pathways could lead to a shear or oxygen regime to direct stem cell differentiation. In this way the efficacy of in vitro culture and differentiation of MSCs on three-dimensional scaffolds could be greatly increased. Bioreactor systems have the ability to control many key differentiation stimuli including mechanical stress and oxygen content. The further integration of cell signaling investigations within dynamic culture systems will lead to a quicker realization of the promise of tissue engineering and regenerative medicine. This article is part of a Special Issue entitled Biochemistry of Stem Cells. Copyright © 2012 Elsevier B.V. All rights reserved.

Sensing in tissue bioreactors

Science.gov (United States)

Rolfe, P.

2006-03-01

Specialized sensing and measurement instruments are under development to aid the controlled culture of cells in bioreactors for the fabrication of biological tissues. Precisely defined physical and chemical conditions are needed for the correct culture of the many cell-tissue types now being studied, including chondrocytes (cartilage), vascular endothelial cells and smooth muscle cells (blood vessels), fibroblasts, hepatocytes (liver) and receptor neurones. Cell and tissue culture processes are dynamic and therefore, optimal control requires monitoring of the key process variables. Chemical and physical sensing is approached in this paper with the aim of enabling automatic optimal control, based on classical cell growth models, to be achieved. Non-invasive sensing is performed via the bioreactor wall, invasive sensing with probes placed inside the cell culture chamber and indirect monitoring using analysis within a shunt or a sampling chamber. Electroanalytical and photonics-based systems are described. Chemical sensing for gases, ions, metabolites, certain hormones and proteins, is under development. Spectroscopic analysis of the culture medium is used for measurement of glucose and for proteins that are markers of cell biosynthetic behaviour. Optical interrogation of cells and tissues is also investigated for structural analysis based on scatter.

Bioreactor design for tendon/ligament engineering.

Science.gov (United States)

Wang, Tao; Gardiner, Bruce S; Lin, Zhen; Rubenson, Jonas; Kirk, Thomas B; Wang, Allan; Xu, Jiake; Smith, David W; Lloyd, David G; Zheng, Ming H

2013-04-01

Tendon and ligament injury is a worldwide health problem, but the treatment options remain limited. Tendon and ligament engineering might provide an alternative tissue source for the surgical replacement of injured tendon. A bioreactor provides a controllable environment enabling the systematic study of specific biological, biochemical, and biomechanical requirements to design and manufacture engineered tendon/ligament tissue. Furthermore, the tendon/ligament bioreactor system can provide a suitable culture environment, which mimics the dynamics of the in vivo environment for tendon/ligament maturation. For clinical settings, bioreactors also have the advantages of less-contamination risk, high reproducibility of cell propagation by minimizing manual operation, and a consistent end product. In this review, we identify the key components, design preferences, and criteria that are required for the development of an ideal bioreactor for engineering tendons and ligaments.

Nano-ceramic composite scaffolds for bioreactor-based bone engineering.

Science.gov (United States)

Lv, Qing; Deng, Meng; Ulery, Bret D; Nair, Lakshmi S; Laurencin, Cato T

2013-08-01

Composites of biodegradable polymers and bioactive ceramics are candidates for tissue-engineered scaffolds that closely match the properties of bone. We previously developed a porous, three-dimensional poly (D,L-lactide-co-glycolide) (PLAGA)/nanohydroxyapatite (n-HA) scaffold as a potential bone tissue engineering matrix suitable for high-aspect ratio vessel (HARV) bioreactor applications. However, the physical and cellular properties of this scaffold are unknown. The present study aims to evaluate the effect of n-HA in modulating PLAGA scaffold properties and human mesenchymal stem cell (HMSC) responses in a HARV bioreactor. By comparing PLAGA/n-HA and PLAGA scaffolds, we asked whether incorporation of n-HA (1) accelerates scaffold degradation and compromises mechanical integrity; (2) promotes HMSC proliferation and differentiation; and (3) enhances HMSC mineralization when cultured in HARV bioreactors. PLAGA/n-HA scaffolds (total number = 48) were loaded into HARV bioreactors for 6 weeks and monitored for mass, molecular weight, mechanical, and morphological changes. HMSCs were seeded on PLAGA/n-HA scaffolds (total number = 38) and cultured in HARV bioreactors for 28 days. Cell migration, proliferation, osteogenic differentiation, and mineralization were characterized at four selected time points. The same amount of PLAGA scaffolds were used as controls. The incorporation of n-HA did not alter the scaffold degradation pattern. PLAGA/n-HA scaffolds maintained their mechanical integrity throughout the 6 weeks in the dynamic culture environment. HMSCs seeded on PLAGA/n-HA scaffolds showed elevated proliferation, expression of osteogenic phenotypic markers, and mineral deposition as compared with cells seeded on PLAGA scaffolds. HMSCs migrated into the scaffold center with nearly uniform cell and extracellular matrix distribution in the scaffold interior. The combination of PLAGA/n-HA scaffolds with HMSCs in HARV bioreactors may allow for the generation of engineered

Generation of Neural Progenitor Spheres from Human Pluripotent Stem Cells in a Suspension Bioreactor.

Science.gov (United States)

Yan, Yuanwei; Song, Liqing; Tsai, Ang-Chen; Ma, Teng; Li, Yan

2016-01-01

Conventional two-dimensional (2-D) culture systems cannot provide large numbers of human pluripotent stem cells (hPSCs) and their derivatives that are demanded for commercial and clinical applications in in vitro drug screening, disease modeling, and potentially cell therapy. The technologies that support three-dimensional (3-D) suspension culture, such as a stirred bioreactor, are generally considered as promising approaches to produce the required cells. Recently, suspension bioreactors have also been used to generate mini-brain-like structure from hPSCs for disease modeling, showing the important role of bioreactor in stem cell culture. This chapter describes a detailed culture protocol for neural commitment of hPSCs into neural progenitor cell (NPC) spheres using a spinner bioreactor. The basic steps to prepare hPSCs for bioreactor inoculation are illustrated from cell thawing to cell propagation. The method for generating NPCs from hPSCs in the spinner bioreactor along with the static control is then described. The protocol in this study can be applied to the generation of NPCs from hPSCs for further neural subtype specification, 3-D neural tissue development, or potential preclinical studies or clinical applications in neurological diseases.

Bioreactors in tissue engineering - principles, applications and commercial constraints.

Science.gov (United States)

Hansmann, Jan; Groeber, Florian; Kahlig, Alexander; Kleinhans, Claudia; Walles, Heike

2013-03-01

Bioreactor technology is vital for tissue engineering. Usually, bioreactors are used to provide a tissue-specific physiological in vitro environment during tissue maturation. In addition to this most obvious application, bioreactors have the potential to improve the efficiency of the overall tissue-engineering concept. To date, a variety of bioreactor systems for tissue-specific applications have been developed. Of these, some systems are already commercially available. With bioreactor technology, various functional tissues of different types were generated and cultured in vitro. Nevertheless, these efforts and achievements alone have not yet led to many clinically successful tissue-engineered implants. We review possible applications for bioreactor systems within a tissue-engineering process and present basic principles and requirements for bioreactor development. Moreover, the use of bioreactor systems for the expansion of clinically relevant cell types is addressed. In contrast to cell expansion, for the generation of functional three-dimensional tissue equivalents, additional physical cues must be provided. Therefore, bioreactors for musculoskeletal tissue engineering are discussed. Finally, bioreactor technology is reviewed in the context of commercial constraints. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Optimising cell aggregate expansion in a perfused hollow fibre bioreactor via mathematical modelling.

KAUST Repository

Chapman, Lloyd A C

2014-08-26

The need for efficient and controlled expansion of cell populations is paramount in tissue engineering. Hollow fibre bioreactors (HFBs) have the potential to meet this need, but only with improved understanding of how operating conditions and cell seeding strategy affect cell proliferation in the bioreactor. This study is designed to assess the effects of two key operating parameters (the flow rate of culture medium into the fibre lumen and the fluid pressure imposed at the lumen outlet), together with the cell seeding distribution, on cell population growth in a single-fibre HFB. This is achieved using mathematical modelling and numerical methods to simulate the growth of cell aggregates along the outer surface of the fibre in response to the local oxygen concentration and fluid shear stress. The oxygen delivery to the cell aggregates and the fluid shear stress increase as the flow rate and pressure imposed at the lumen outlet are increased. Although the increased oxygen delivery promotes growth, the higher fluid shear stress can lead to cell death. For a given cell type and initial aggregate distribution, the operating parameters that give the most rapid overall growth can be identified from simulations. For example, when aggregates of rat cardiomyocytes that can tolerate shear stresses of up to 0:05 Pa are evenly distributed along the fibre, the inlet flow rate and outlet pressure that maximise the overall growth rate are predicted to be in the ranges 2.75 x 10(-5) m(2) s(-1) to 3 x 10(-5) m(2) s(-1) (equivalent to 2.07 ml min(-1) to 2.26 ml min(-1)) and 1.077 x 10(5) Pa to 1.083 x 10(5) Pa (or 15.6 psi to 15.7 psi) respectively. The combined effects of the seeding distribution and flow on the growth are also investigated and the optimal conditions for growth found to depend on the shear tolerance and oxygen demands of the cells.

Simulation of temperature effect on microalgae culture in a tubular photo bioreactor for local solar irradiance

Science.gov (United States)

Shahriar, M.; Deb, Ujjwal Kumar; Rahman, Kazi Afzalur

2017-06-01

Microalgae based biofuel is now an emerging source of renewable energy alternative to the fossil fuel. This paper aims to present computational model of microalgae culture taking effect of solar irradiance and corresponding temperature in a photo bioreactor (PBR). As microalgae is a photosynthetic microorganism, so irradiance of sunlight is one of the important limiting factors for the proper growth of microalgae cells as temperature is associated with it. We consider the transient behaviour of temperature inside the photo bioreactor for a microalgae culture. The optimum range of temperature for outdoor cultivation of microalgae is about 16-35°c and out of this range the cell growth inhibits. Many correlations have already been established to investigate the heat transfer phenomena inside a tubular PBR. However, none of them are validated yet numerically by using a user defined function in a simulated model. A horizontal tubular PBR length 20.5m with radius 0.05m has taken account to investigate the temperature effect for the growth of microalgae cell. As the solar irradiance varies at any geographic latitude for a year so an empirical relation is established between local solar irradiance and temperature to simulate the effect. From our simulation, we observed that the growth of microalgae has a significant effect of temperature and the solar irradiance of our locality is suitable for the culture of microalgae.

Construction and evaluation of urinary bladder bioreactor for urologic tissue-engineering purposes.

LENUS (Irish Health Repository)

Davis, Niall F

2012-01-31

OBJECTIVE: To design and construct a urinary bladder bioreactor for urologic tissue-engineering purposes and to compare the viability and proliferative activity of cell-seeded extracellular matrix scaffolds cultured in the bioreactor with conventional static growth conditions. MATERIALS AND METHODS: A urinary bladder bioreactor was designed and constructed to replicate physiologic bladder dynamics. The bioreactor mimicked the filling pressures of the human bladder by way of a cyclical low-delivery pressure regulator. In addition, cell growth was evaluated by culturing human urothelial cells (UCs) on porcine extracellular matrix scaffolds in the bioreactor and in static growth conditions for 5 consecutive days. The attachment, viability, and proliferative potential were assessed and compared with quantitative viability indicators and by fluorescent markers for intracellular esterase activity and plasma membrane integrity. Scaffold integrity was characterized with scanning electron microscopy and 4\\

Repair of segmental bone defect using Totally Vitalized tissue engineered bone graft by a combined perfusion seeding and culture system.

Directory of Open Access Journals (Sweden)

Lin Wang

Full Text Available BACKGROUND: The basic strategy to construct tissue engineered bone graft (TEBG is to combine osteoblastic cells with three dimensional (3D scaffold. Based on this strategy, we proposed the "Totally Vitalized TEBG" (TV-TEBG which was characterized by abundant and homogenously distributed cells with enhanced cell proliferation and differentiation and further investigated its biological performance in repairing segmental bone defect. METHODS: In this study, we constructed the TV-TEBG with the combination of customized flow perfusion seeding/culture system and β-tricalcium phosphate (β-TCP scaffold fabricated by Rapid Prototyping (RP technique. We systemically compared three kinds of TEBG constructed by perfusion seeding and perfusion culture (PSPC method, static seeding and perfusion culture (SSPC method, and static seeding and static culture (SSSC method for their in vitro performance and bone defect healing efficacy with a rabbit model. RESULTS: Our study has demonstrated that TEBG constructed by PSPC method exhibited better biological properties with higher daily D-glucose consumption, increased cell proliferation and differentiation, and better cell distribution, indicating the successful construction of TV-TEBG. After implanted into rabbit radius defects for 12 weeks, PSPC group exerted higher X-ray score close to autograft, much greater mechanical property evidenced by the biomechanical testing and significantly higher new bone formation as shown by histological analysis compared with the other two groups, and eventually obtained favorable healing efficacy of the segmental bone defect that was the closest to autograft transplantation. CONCLUSION: This study demonstrated the feasibility of TV-TEBG construction with combination of perfusion seeding, perfusion culture and RP technique which exerted excellent biological properties. The application of TV-TEBG may become a preferred candidate for segmental bone defect repair in orthopedic and

Feasibility of using sodium chloride as a tracer for the characterization of the distribution of matter in complex multi-compartment 3D bioreactors for stem cell culture.

Science.gov (United States)

Gerlach, Jörg C; Witaschek, Tom; Strobel, Catrin; Brayfield, Candace A; Bornemann, Reinhard; Catapano, Gerardo; Zeilinger, Katrin

2010-06-01

The experimental characterization of the distribution of matter in complex multi-compartment three-dimensional membrane bioreactors for human cell culture is complicated by tracer interactions with the membranes and other bioreactor constituents. This is due to the fact that membranes with a high specific surface area often feature a hydrophobic chemical backbone that may adsorb tracers often used to this purpose, such as proteins and dyes. Membrane selectivity, and its worsening caused by protein adsorption, may also hinder tracer transfer across neighboring compartments, thus preventing effective characterization of the distribution of matter in the whole bioreactor. Tracer experiments with sodium chloride (NaCl) may overcome some of these limitations and be effectively used to characterize the distribution of matter in complex 3D multi-compartments membrane bioreactors for stem cell culture. NaCl freely permeates most used membranes, it does not adsorb on uncharged membranes, and its concentration may be accurately measured in terms of solution conductivity. In this preliminary study, the feasibility of complex multi-compartment membrane bioreactors was investigated with a NaCl concentration pulse challenge to characterize how their distribution of matter changes when they are operated under different conditions. In particular, bioreactors consisting of three different membrane types stacked on top of one another to form a 3D network were characterized under different feed conditions.

Oxygen Limited Bioreactors System For Nitrogen Removal Using Immobilized Mix Culture

Science.gov (United States)

Pathak, B. K.; Sumino, T.; Saiki, Y.; Kazama, F.

2005-12-01

Recently nutrients concentrations especially nitrogen in natural water is alarming in the world wide. Most of the effort is being done on the removal of high concentration of nitrogen especially from the wastewater treatment plants. The removal efficiency is targeted in all considering the effluent discharge standard set by the national environment agency. In many cases, it does not meet the required standard and receiving water is being polluted. Eutrophication in natural water bodies has been reported even if the nitrogen concentration is low and self purification of natural systems itself is not sufficient to remove the nitrogen due to complex phenomenon. In order to recover the pristine water environment, it is very essential to explore bioreactor systems for natural water systems using immobilized mix culture. Microorganism were entrapped in Polyethylene glycol (PEG) prepolymer gel and cut into 3mm cubic immobilized pellets. Four laboratory scale micro bio-reactors having 0.1 L volumes were packed with immobilized pellets with 50% compact ratio. RUN1, RUN2, RUN3 and RUN4 were packed with immobilized pellets from reservoirs sediments, activated sludge (AS), mixed of AS, AG and biodegradable plastic and anaerobic granules (AG) respectively. Water from Shiokawa Reservoirs was feed to all reactors with supplemental ammonia and nitrite nitrogen as specified in the results and discussions. The reactors were operated dark incubated room in continuous flow mode with hydraulic retention time of 12 hours under oxygen limiting condition. Ammonium, nitrate nitrite nitrogen and total organic carbon (TOC) concentrations were measured as described in APWA and AWWA (1998). Laboratory scale four bioreactors containing different combination of immobilized cell were monitored for 218 days. Influent NH4+-N and NO2--N concentration were 2.27±0.43 and 2.05±0.41 mg/l respectively. Average dissolved oxygen concentration and pH in the reactors were 0.40-2.5 mg/l and pH 6

Universal lab-on-a-chip platform for complex, perfused 3D cell cultures

Science.gov (United States)

Sonntag, F.; Schmieder, F.; Ströbel, J.; Grünzner, S.; Busek, M.; Günther, K.; Steege, T.; Polk, C.; Klotzbach, U.

2016-03-01

The miniaturization, rapid prototyping and automation of lab-on-a-chip technology play nowadays a very important role. Lab-on-a-chip technology is successfully implemented not only for environmental analysis and medical diagnostics, but also as replacement of animals used for the testing of substances in the pharmaceutical and cosmetics industries. For that purpose the Fraunhofer IWS and partners developed a lab-on-a-chip platform for perfused cell-based assays in the last years, which includes different micropumps, valves, channels, reservoirs and customized cell culture modules. This technology is already implemented for the characterization of different human cell cultures and organoids, like skin, liver, endothelium, hair follicle and nephron. The advanced universal lab-on-a-chip platform for complex, perfused 3D cell cultures is divided into a multilayer basic chip with integrated micropump and application-specific 3D printed cell culture modules. Moreover a technology for surface modification of the printed cell culture modules by laser micro structuring and a complex and flexibly programmable controlling device based on an embedded Linux system was developed. A universal lab-on-a-chip platform with an optional oxygenator and a cell culture module for cubic scaffolds as well as first cell culture experiments within the cell culture device will be presented. The module is designed for direct interaction with robotic dispenser systems. This offers the opportunity to combine direct organ printing of cells and scaffolds with the microfluidic cell culture module. The characterization of the developed system was done by means of Micro-Particle Image Velocimetry (μPIV) and an optical oxygen measuring system.

Construction and characterization of a novel vocal fold bioreactor.

Science.gov (United States)

Zerdoum, Aidan B; Tong, Zhixiang; Bachman, Brendan; Jia, Xinqiao

2014-08-01

In vitro engineering of mechanically active tissues requires the presentation of physiologically relevant mechanical conditions to cultured cells. To emulate the dynamic environment of vocal folds, a novel vocal fold bioreactor capable of producing vibratory stimulations at fundamental phonation frequencies is constructed and characterized. The device is composed of a function generator, a power amplifier, a speaker selector and parallel vibration chambers. Individual vibration chambers are created by sandwiching a custom-made silicone membrane between a pair of acrylic blocks. The silicone membrane not only serves as the bottom of the chamber but also provides a mechanism for securing the cell-laden scaffold. Vibration signals, generated by a speaker mounted underneath the bottom acrylic block, are transmitted to the membrane aerodynamically by the oscillating air. Eight identical vibration modules, fixed on two stationary metal bars, are housed in an anti-humidity chamber for long-term operation in a cell culture incubator. The vibration characteristics of the vocal fold bioreactor are analyzed non-destructively using a Laser Doppler Vibrometer (LDV). The utility of the dynamic culture device is demonstrated by culturing cellular constructs in the presence of 200-Hz sinusoidal vibrations with a mid-membrane displacement of 40 µm. Mesenchymal stem cells cultured in the bioreactor respond to the vibratory signals by altering the synthesis and degradation of vocal fold-relevant, extracellular matrix components. The novel bioreactor system presented herein offers an excellent in vitro platform for studying vibration-induced mechanotransduction and for the engineering of functional vocal fold tissues.

Microfluidic culture chamber for the long-term perfusion and precise chemical stimulation of organotypic brain tissue slices

DEFF Research Database (Denmark)

Caicedo, H. H.; Vignes, M.; Brugg, B.

2010-01-01

We have developed a microfluidic perfusion-based culture system to study long-term in-vitro responses of organo-typic brain slices exposed to localized neurochemical stimulation. Using this microperfusion chamber we show that hip-pocampal organotypic brain slices cultures grown on nitrocellulose ...

3D perfusion bioreactor-activated porous granules on implant fixation and early bone formation in sheep

DEFF Research Database (Denmark)

Ding, Ming; Snoek Henriksen, Susan; Martinetti, Roberta

2017-01-01

allograft, granules, granules with bone marrow aspirate or bioreactor-activated graft material. Following an observation time of 6 weeks, early implant fixation and bone formation were assessed by micro-CT scanning, mechanical testing, and histomorphometry. Bone formations were seen in all groups, while......, bone formation was observed in all groups, while the bioreactor-activated graft material did not reveal additional effects on early implant fixation comparable to allograft in this model. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2016....

Transparent polymeric cell culture chip with integrated temperature control and uniform media perfusion

DEFF Research Database (Denmark)

Petronis, Sarunas; Stangegaard, Michael; Christensen, C.

2006-01-01

Modern microfabrication and microfluidic technologies offer new opportunities in the design and fabrication of miniaturized cell culture systems for online monitoring of living cells. We used laser micromachining and thermal bonding to fabricate an optically transparent, low-cost polymeric chip...... for long-term online cell culture observation under controlled conditions. The chip incorporated a microfluidic flow equalization system, assuring uniform perfusion of the cell culture media throughout the cell culture chamber. The integrated indium-tin-oxide heater and miniature temperature probe linked....... HeLa cells were cultured for up to 2 weeks within the cell culture chip and monitored using a time-lapse video recording microscopy setup. Cell attachment and spreading was observed during the first 10-20 h (lag phase). After approximately 20 h, cell growth gained exponential character...

Growth of plant root cultures in liquid- and gas-dispersed reactor environments.

Science.gov (United States)

McKelvey, S A; Gehrig, J A; Hollar, K A; Curtis, W R

1993-01-01

The growth of Agrobacterium transformed "hairy root" cultures of Hyoscyamus muticus was examined in various liquid- and gas-dispersed bioreactor configurations. Reactor runs were replicated to provide statistical comparisons of nutrient availability on culture performance. Accumulated tissue mass in submerged air-sparged reactors was 31% of gyratory shake-flask controls. Experiments demonstrate that poor performance of sparged reactors is not due to bubble shear damage, carbon dioxide stripping, settling, or flotation of roots. Impaired oxygen transfer due to channeling and stagnation of the liquid phase are the apparent causes of poor growth. Roots grown on a medium-perfused inclined plane grew at 48% of gyratory controls. This demonstrates the ability of cultures to partially compensate for poor liquid distribution through vascular transport of nutrients. A reactor configuration in which the medium is sprayed over the roots and permitted to drain down through the root tissue was able to provide growth rates which are statistically indistinguishable (95% T-test) from gyratory shake-flask controls. In this type of spray/trickle-bed configuration, it is shown that distribution of the roots becomes a key factor in controlling the rate of growth. Implications of these results regarding design and scale-up of bioreactors to produce fine chemicals from root cultures are discussed.

A recapitulative three-dimensional model of breast carcinoma requires perfusion for multi-week growth

Directory of Open Access Journals (Sweden)

Kayla F Goliwas

2016-07-01

Full Text Available Breast carcinomas are complex, three-dimensional tissues composed of cancer epithelial cells and stromal components, including fibroblasts and extracellular matrix. In vitro models that more faithfully recapitulate this dimensionality and stromal microenvironment should more accurately elucidate the processes driving carcinogenesis, tumor progression, and therapeutic response. Herein, novel in vitro breast carcinoma surrogates, distinguished by a relevant dimensionality and stromal microenvironment, are described and characterized. A perfusion bioreactor system was used to deliver medium to surrogates containing engineered microchannels and the effects of perfusion, medium composition, and the method of cell incorporation and density of initial cell seeding on the growth and morphology of surrogates were assessed. Perfused surrogates demonstrated significantly greater cell density and proliferation and were more histologically recapitulative of human breast carcinoma than surrogates maintained without perfusion. Although other parameters of the surrogate system, such as medium composition and cell seeding density, affected cell growth, perfusion was the most influential parameter.

High-yield secretion of recombinant proteins expressed in tobacco cell culture with a designer glycopeptide tag: Process development.

Science.gov (United States)

Zhang, Ningning; Gonzalez, Maria; Savary, Brett; Xu, Jianfeng

2016-03-01

Low-yield protein production remains the most significant economic hurdle with plant cell culture technology. Fusions of recombinant proteins with hydroxyproline-O-glycosylated designer glycopeptide tags have consistently boosted secreted protein yields. This prompted us to study the process development of this technology aiming to achieve productivity levels necessary for commercial viability. We used a tobacco BY-2 cell culture expressing EGFP as fusion with a glycopeptide tag comprised of 32 repeat of "Ser-Pro" dipeptide, or (SP)32 , to study cell growth and protein secretion, culture scale-up, and establishment of perfusion cultures for continuous production. The BY-2 cells accumulated low levels of cell biomass (~7.5 g DW/L) in Schenk & Hildebrandt medium, but secreted high yields of (SP)32 -tagged EGFP (125 mg/L). Protein productivity of the cell culture has been stable for 6.0 years. The BY-2 cells cultured in a 5-L bioreactor similarly produced high secreted protein yield at 131 mg/L. Successful operation of a cell perfusion culture for 30 days was achieved under the perfusion rate of 0.25 and 0.5 day(-1) , generating a protein volumetric productivity of 17.6 and 28.9 mg/day/L, respectively. This research demonstrates the great potential of the designer glycopeptide technology for use in commercial production of valuable proteins with plant cell cultures. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Mechanobiologic Research in a Microgravity Environment Bioreactor

Science.gov (United States)

Guidi, A.; Dubini, G.; Tominetti, F.; Raimondi, M.

A current problem in tissue culturing technology is the unavailability of an effective Bioreactor for the in vitro cultivation of cells and explants. It has, in fact, proved extremely difficult to promote the high-density three-dimensional in vitro growth of human tissues that have been removed from the body and deprived of their normal in vivo vascular sources of nutrients and gas exchange. A variety of tissue explants can be maintained for a short period of time on a supportive collagen matrix surrounded by culture medium. But this system provides only limited mass transfer of nutrients and wastes through the tissue, and gravity-induced sedimentation prevents complete three- dimensional cell-cell and cell-matrix interactions. Several devices presently on the market have been used with only limited success since each has limitations, which restrict usefulness and versatility. Further, no Bioreactor or culture vessel is known that will allow for unimpeded growth of three dimensional cellular aggregates or tissue. Extensive research on the effect of mechanical stimuli on cell metabolism suggests that tissues may respond to mechanical stimulation via loading-induced flow of the interstitial fluids. During the culture, cells are subject to a flow of culture medium. Flow properties such as flow field, flow regime (e.g. turbulent or laminar), flow pattern (e.g. circular), entity and distribution of the shear stress acting on the cells greatly influence fundamental aspects of cell function, such as regulation and gene expression. This has been demonstrated for endothelial cells and significant research efforts are underway to elucidate these mechanisms in various other biological systems. Local fluid dynamics is also responsible of the mass transfer of nutrients and catabolites as well as oxygenation through the tissue. Most of the attempts to culture tissue-engineered constructs in vitro have utilized either stationary cultures or systems generating relatively small

In Vitro Model for Hepatotoxicity Studies Based on Primary Human Hepatocyte Cultivation in a Perfused 3D Bioreactor System.

Science.gov (United States)

Knöspel, Fanny; Jacobs, Frank; Freyer, Nora; Damm, Georg; De Bondt, An; van den Wyngaert, Ilse; Snoeys, Jan; Monshouwer, Mario; Richter, Marco; Strahl, Nadja; Seehofer, Daniel; Zeilinger, Katrin

2016-04-16

Accurate prediction of the potential hepatotoxic nature of new pharmaceuticals remains highly challenging. Therefore, novel in vitro models with improved external validity are needed to investigate hepatic metabolism and timely identify any toxicity of drugs in humans. In this study, we examined the effects of diclofenac, as a model substance with a known risk of hepatotoxicity in vivo, in a dynamic multi-compartment bioreactor using primary human liver cells. Biotransformation pathways of the drug and possible effects on metabolic activities, morphology and cell transcriptome were evaluated. Formation rates of diclofenac metabolites were relatively stable over the application period of seven days in bioreactors exposed to 300 µM diclofenac (300 µM bioreactors (300 µM BR)), while in bioreactors exposed to 1000 µM diclofenac (1000 µM BR) metabolite concentrations declined drastically. The biochemical data showed a significant decrease in lactate production and for the higher dose a significant increase in ammonia secretion, indicating a dose-dependent effect of diclofenac application. The microarray analyses performed revealed a stable hepatic phenotype of the cells over time and the observed transcriptional changes were in line with functional readouts of the system. In conclusion, the data highlight the suitability of the bioreactor technology for studying the hepatotoxicity of drugs in vitro.

In Vitro Model for Hepatotoxicity Studies Based on Primary Human Hepatocyte Cultivation in a Perfused 3D Bioreactor System

Directory of Open Access Journals (Sweden)

Fanny Knöspel

2016-04-01

Full Text Available Accurate prediction of the potential hepatotoxic nature of new pharmaceuticals remains highly challenging. Therefore, novel in vitro models with improved external validity are needed to investigate hepatic metabolism and timely identify any toxicity of drugs in humans. In this study, we examined the effects of diclofenac, as a model substance with a known risk of hepatotoxicity in vivo, in a dynamic multi-compartment bioreactor using primary human liver cells. Biotransformation pathways of the drug and possible effects on metabolic activities, morphology and cell transcriptome were evaluated. Formation rates of diclofenac metabolites were relatively stable over the application period of seven days in bioreactors exposed to 300 µM diclofenac (300 µM bioreactors (300 µM BR, while in bioreactors exposed to 1000 µM diclofenac (1000 µM BR metabolite concentrations declined drastically. The biochemical data showed a significant decrease in lactate production and for the higher dose a significant increase in ammonia secretion, indicating a dose-dependent effect of diclofenac application. The microarray analyses performed revealed a stable hepatic phenotype of the cells over time and the observed transcriptional changes were in line with functional readouts of the system. In conclusion, the data highlight the suitability of the bioreactor technology for studying the hepatotoxicity of drugs in vitro.

A mini-scale mass production and separation system for secretory heterologous proteins by perfusion culture of recombinant Pichia pastoris using a shaken ceramic membrane flask.

Science.gov (United States)

Ohashi, R; Mochizuki, E; Suzuki, T

1999-01-01

The perfusion culture technique using a shaken ceramic membrane flask (SCM flask) was applied to the production of a secretory heterologous protein. A recombinant methylotrophic yeast strain, Pichia pastoris, was cultured aerobically on a reciprocal shaker using an SCM flask. High-level production of human serum albumin (HSA) was attempted by increasing both the cell concentration and the expression level of the recombinant gene. In the two-stage culture method, the cell concentration was first raised to 17 g/l by feeding glycerol, after which the expression of HSA was induced by feeding methanol. However, the concentration of HSA in the effluent filtrate was as low as 0.15 g/l, while the cell concentration continued to increase. In contrast, HSA was effectively produced by feeding methanol from an early stage of the culture. In this case, the HSA concentration reached 0.24 and 0.46 g/l, respectively, using the growth-associated production method without and with aeration into the head space of the SCM flask. The results showed that supplying sufficient oxygen together with the growth-associated induction method are effective for obtaining high-level expression of the methanol-inducible recombinant gene of P. pastoris. An HSA concentration in the filtrate of 1.5 g/l was finally achieved when the cell concentration was increased to 53 g/l by supplying oxygen-enriched gas to the SCM flask. The yield and productivity of HSA reached 2.6-fold and 10-fold those obtained in an ordinary fed-batch culture using a shake flask, and these levels were readily achieved by continuous replenishment of the culture supernatant. The achievements made in this study should contribute to the development of a handy bioreactor system for mini-scale mass production of target proteins with separation at high purity.

Preparation of kombucha from winter savory (Satureja Montana L. in the laboratory bioreactor

Directory of Open Access Journals (Sweden)

Cvetković Dragoljub D.

2005-01-01

Full Text Available The possibility of obtaining kombucha from winter savory tea has been tested in the laboratory bioreactor by applying starter cultures and traditional way of inoculation. On the basis of the obtained results, it can be concluded that applying the inoculating method with the beverage from the previous process of biotransformation yielded kombucha beverage (capacity 15 I from winter savory tea in the laboratory bioreactor. The application of defined starter culture from the isolate of yeast and acetic acid bacteria of local tea in the glass jar (capacity 5 I gave 3 litres of kombucha beverage, which is acceptable according to the basic parameters and sensory characteristics. However, the application of the same starter culture in the laboratory bioreactor did not result in synchronized activity of yeast and bacteria.

Simultaneous environmental manipulations in semi-perfusion cultures of CHO cells producing rh-tPA

OpenAIRE

Vergara,Mauricio; Becerra,Silvana; Díaz-Barrera,Alvaro; Berrios,Julio; Altamirano,Claudia

2012-01-01

We evaluated the combined effect of decreasing the temperature to a mild hypothermia range (34 and 31ºC) and switching to a slowly metabolizable carbon source (glucose substituted by galactose) on the growth and production of a recombinant human tissue plasminogen activator (rh-tPA) by Chinese hamster ovary cells in batch and semi-perfusion cultures. In batch cultures using glucose as a carbon source, decreasing the temperature caused a reduction in cell growth and an increase in specific pro...

Bioreactor activated graft material for early implant fixation in bone

DEFF Research Database (Denmark)

Snoek Henriksen, Susan; Ding, Ming; Overgaard, Søren

2011-01-01

from the iliac crest. For both groups, mononuclear cells were isolated, and injected into a perfusion bioreactor (Millenium Biologix AG, Switzerland). Scaffold granules (Ø~900-1500 µm, ~88% porosity) in group 1, consisted of hydroxyapatite (HA, 70%) with β-tricalcium-phosphate (β-TCP, 30%) (Danish....... The superficial part was used for mechanical testing and micro-CT scanning, and the profound part for histomorphometry. Push-out tests were performed on an 858 Bionix MTS hydraulic materials testing machine (MTS Systems Corporation, USA). Shear mechanical properties between implant and newly generated bone were...

Biomimetic fetal rotation bioreactor for engineering bone tissues-Effect of cyclic strains on upregulation of osteogenic gene expression.

Science.gov (United States)

Ravichandran, Akhilandeshwari; Wen, Feng; Lim, Jing; Chong, Mark Seow Khoon; Chan, Jerry K Y; Teoh, Swee-Hin

2018-04-01

Cells respond to physiological mechanical stresses especially during early fetal development. Adopting a biomimetic approach, it is necessary to develop bioreactor systems to explore the effects of physiologically relevant mechanical strains and shear stresses for functional tissue growth and development. This study introduces a multimodal bioreactor system that allows application of cyclic compressive strains on premature bone grafts that are cultured under biaxial rotation (chamber rotation about 2 axes) conditions for bone tissue engineering. The bioreactor is integrated with sensors for dissolved oxygen levels and pH that allow real-time, non-invasive monitoring of the culture parameters. Mesenchymal stem cells-seeded polycaprolactone-β-tricalcium phosphate scaffolds were cultured in this bioreactor over 2 weeks in 4 different modes-static, cyclic compression, biaxial rotation, and multimodal (combination of cyclic compression and biaxial rotation). The multimodal culture resulted in 1.8-fold higher cellular proliferation in comparison with the static controls within the first week. Two weeks of culture in the multimodal bioreactor utilizing the combined effects of optimal fluid flow conditions and cyclic compression led to the upregulation of osteogenic genes alkaline phosphatase (3.2-fold), osteonectin (2.4-fold), osteocalcin (10-fold), and collagen type 1 α1 (2-fold) in comparison with static cultures. We report for the first time, the independent and combined effects of mechanical stimulation and biaxial rotation for bone tissue engineering using a bioreactor platform with non-invasive sensing modalities. The demonstrated results show leaning towards the futuristic vision of using a physiologically relevant bioreactor system for generation of autologous bone grafts for clinical implantation. Copyright © 2018 John Wiley & Sons, Ltd.

Cyclic Loading of Growing Tissue in a Bioreactor: Mathematical Model and Asymptotic Analysis

KAUST Repository

Pohlmeyer, J. V.

2013-10-24

A simplified 2D mathematical model for tissue growth within a cyclically-loaded tissue engineering scaffold is presented and analyzed. Such cyclic loading has the potential to improve yield and functionality of tissue such as bone and cartilage when grown on a scaffold within a perfusion bioreactor. The cyclic compression affects the flow of the perfused nutrient, leading to flow properties that are inherently unsteady, though periodic, on a timescale short compared with that of tissue proliferation. A two-timescale analysis based on these well-separated timescales is exploited to derive a closed model for the tissue growth on the long timescale of proliferation. Some sample numerical results are given for the final model, and discussed. © 2013 Society for Mathematical Biology.

Trickle-bed root culture bioreactor design and scale-up: growth, fluid-dynamics, and oxygen mass transfer.

Science.gov (United States)

Ramakrishnan, Divakar; Curtis, Wayne R

2004-10-20

Trickle-bed root culture reactors are shown to achieve tissue concentrations as high as 36 g DW/L (752 g FW/L) at a scale of 14 L. Root growth rate in a 1.6-L reactor configuration with improved operational conditions is shown to be indistinguishable from the laboratory-scale benchmark, the shaker flask (mu=0.33 day(-1)). These results demonstrate that trickle-bed reactor systems can sustain tissue concentrations, growth rates and volumetric biomass productivities substantially higher than other reported bioreactor configurations. Mass transfer and fluid dynamics are characterized in trickle-bed root reactors to identify appropriate operating conditions and scale-up criteria. Root tissue respiration goes through a minimum with increasing liquid flow, which is qualitatively consistent with traditional trickle-bed performance. However, liquid hold-up is much higher than traditional trickle-beds and alternative correlations based on liquid hold-up per unit tissue mass are required to account for large changes in biomass volume fraction. Bioreactor characterization is sufficient to carry out preliminary design calculations that indicate scale-up feasibility to at least 10,000 liters.

Transfection in perfused microfluidic cell culture devices: A case study.

Science.gov (United States)

Raimes, William; Rubi, Mathieu; Super, Alexandre; Marques, Marco P C; Veraitch, Farlan; Szita, Nicolas

2017-08-01

Automated microfluidic devices are a promising route towards a point-of-care autologous cell therapy. The initial steps of induced pluripotent stem cell (iPSC) derivation involve transfection and long term cell culture. Integration of these steps would help reduce the cost and footprint of micro-scale devices with applications in cell reprogramming or gene correction. Current examples of transfection integration focus on maximising efficiency rather than viable long-term culture. Here we look for whole process compatibility by integrating automated transfection with a perfused microfluidic device designed for homogeneous culture conditions. The injection process was characterised using fluorescein to establish a LabVIEW-based routine for user-defined automation. Proof-of-concept is demonstrated by chemically transfecting a GFP plasmid into mouse embryonic stem cells (mESCs). Cells transfected in the device showed an improvement in efficiency (34%, n = 3) compared with standard protocols (17.2%, n = 3). This represents a first step towards microfluidic processing systems for cell reprogramming or gene therapy.

Aujeszky's disease virus production in disposable bioreactor

Indian Academy of Sciences (India)

Madhu

1Laboratory for Cell Culture Technology and Biotransformations, 2Laboratory for ... A novel, disposable-bag bioreactor system that uses wave action for mixing and transferring ... consisted of 95% of air + 5% of CO2 using gas mixing module.

Biomimetic Culture Reactor for Whole-Lung Engineering.

Science.gov (United States)

Raredon, Micha Sam Brickman; Rocco, Kevin A; Gheorghe, Ciprian P; Sivarapatna, Amogh; Ghaedi, Mahboobe; Balestrini, Jenna L; Raredon, Thomas L; Calle, Elizabeth A; Niklason, Laura E

2016-01-01

Decellularized organs are now established as promising scaffolds for whole-organ regeneration. For this work to reach therapeutic practice, techniques and apparatus are necessary for doing human-scale clinically applicable organ cultures. We have designed and constructed a bioreactor system capable of accommodating whole human or porcine lungs, and we describe in this study relevant technical details, means of assembly and operation, and validation. The reactor has an artificial diaphragm that mimics the conditions found in the chest cavity in vivo, driving hydraulically regulated negative pressure ventilation and custom-built pulsatile perfusion apparatus capable of driving pressure-regulated or volume-regulated vascular flow. Both forms of mechanical actuation can be tuned to match specific physiologic profiles. The organ is sealed in an elastic artificial pleura that mounts to a support architecture. This pleura reduces the fluid volume required for organ culture, maintains the organ's position during mechanical conditioning, and creates a sterile barrier allowing disassembly and maintenance outside of a biosafety cabinet. The combination of fluid suspension, negative-pressure ventilation, and physiologic perfusion allows the described system to provide a biomimetic mechanical environment not found in existing technologies and especially suited to whole-organ regeneration. In this study, we explain the design and operation of this apparatus and present data validating intended functions.

Bioreactor cultivation enhances NTEB formation and differentiation of NTES cells into cardiomyocytes.

Science.gov (United States)

Lü, Shuanghong; Liu, Sheng; He, Wenjun; Duan, Cuimi; Li, Yanmin; Liu, Zhiqiang; Zhang, Ye; Hao, Tong; Wang, Yanmeng; Li, Dexue; Wang, Changyong; Gao, Shaorong

2008-09-01

Autogenic embryonic stem cells established from somatic cell nuclear transfer (SCNT) embryos have been proposed as unlimited cell sources for cell transplantation-based treatment of many genetic and degenerative diseases, which can eliminate the immune rejection that occurs after transplantation. In the present study, pluripotent nuclear transfer ES (NTES) cell lines were successfully established from different strains of mice. One NTES cell line, NT1, with capacity of germline transmission, was used to investigate in vitro differentiation into cardiomyocytes. To optimize differentiation conditions for mass production of embryoid bodies (NTEBs) from NTES cells, a slow-turning lateral vessel (STLV) rotating bioreactor was used for culturing the NTES cells to produce NTEBs compared with a conventional static cultivation method. Our results demonstrated that the NTEBs formed in STLV bioreactor were more uniform in size, and no large necrotic centers with most of the cells in NTEBs were viable. Differentiation of the NTEBs formed in both the STLV bioreactor and static culture into cardiomyocytes was induced by ascorbic acid, and the results demonstrated that STLV-produced NTEBs differentiated into cardiomyocytes more efficiently. Taken together, our results suggested that STLV bioreactor provided a more ideal culture condition, which can facilitate the formation of better quality NTEBs and differentiation into cardiomyocytes more efficiently in vitro.

Commissioning of Research Bioreactor made in Korea with Malaysian Environment Adaptation

International Nuclear Information System (INIS)

Mohd Jamil Hashim; Mohd Azmi Sidid Omar

2011-01-01

Bioreactor is equipment used by researcher in Agrotechnology and Biosciences department (BAB) as a scientific approach to get a scale up of product. Headed by one of the senior researcher in the department, an effort has been made to upscale the project by using MTDC fund. The technology platform has been acquired from South Korea. Some modification has to be made to cater for the need of a research bioreactor to be established for Nuclear Malaysia Agency. This research bioreactor is to emulate a tissue culture product in a bigger scale bio processing, pharmaceutical biotechnology and industrial production. (author)

Technical and theoretical considerations about gradient perfusion culture for epithelia used in tissue engineering, biomaterial testing and pharmaceutical research

International Nuclear Information System (INIS)

Minuth, Will W; Strehl, Raimund

2007-01-01

Epithelia act as biological barriers, which are exposed to different environments at the luminal and basal sides. To simulate this situation and to improve functional features an in vitro gradient perfusion culture technique was developed in our laboratory. This innovative technique appears to be simple at first sight, but the performance needs practical and theoretical knowledge. To harvest intact epithelia after a long-term gradient culture period of many days, leakage, edge damage and pressure differences in the system have to be avoided so that the epithelial barrier function is maintained continuously. Unexpectedly, one of the major obstacles are micro-injuries in the epithelia caused by gas bubbles, which arise during transportation of the medium or due to respiration of the cultured tissue. Gas bubbles randomly accumulate either at the luminal or basal fluid flow of the gradient perfusion culture container. This phenomenon results in fluid pressure differences between the luminal and basal perfusion compartments of the gradient container, which in turn leads to damage of the barrier function. Consequently, the content of gas bubbles in the transported culture medium has to be minimized. Thus, our technical concept is the reduction of gas bubbles while keeping the content of oxygen constant. To follow this strategy we developed a new type of screw cap for media bottles specifically designed to allow fluid contact only with tube and not with cap material. Furthermore, a gas expander module separates gas bubbles from the liquid phase during transportation of the medium. Finally, a new type of gradient culture container allows a permanent elimination of transported gas bubbles. Application of this innovative equipment optimizes the parallel transportation of fluid in the luminal and basal compartments of a gradient culture container. (topical review)

Technical and theoretical considerations about gradient perfusion culture for epithelia used in tissue engineering, biomaterial testing and pharmaceutical research

Energy Technology Data Exchange (ETDEWEB)

Minuth, Will W [Department of Molecular and Cellular Anatomy, University of Regensburg, D-93053 Regensburg, University Street 31 (Germany); Strehl, Raimund [Cellartis AB, S-41346 Goeteborg, Arvid Wallgrens Backe 20 (Sweden)

2007-06-01

Epithelia act as biological barriers, which are exposed to different environments at the luminal and basal sides. To simulate this situation and to improve functional features an in vitro gradient perfusion culture technique was developed in our laboratory. This innovative technique appears to be simple at first sight, but the performance needs practical and theoretical knowledge. To harvest intact epithelia after a long-term gradient culture period of many days, leakage, edge damage and pressure differences in the system have to be avoided so that the epithelial barrier function is maintained continuously. Unexpectedly, one of the major obstacles are micro-injuries in the epithelia caused by gas bubbles, which arise during transportation of the medium or due to respiration of the cultured tissue. Gas bubbles randomly accumulate either at the luminal or basal fluid flow of the gradient perfusion culture container. This phenomenon results in fluid pressure differences between the luminal and basal perfusion compartments of the gradient container, which in turn leads to damage of the barrier function. Consequently, the content of gas bubbles in the transported culture medium has to be minimized. Thus, our technical concept is the reduction of gas bubbles while keeping the content of oxygen constant. To follow this strategy we developed a new type of screw cap for media bottles specifically designed to allow fluid contact only with tube and not with cap material. Furthermore, a gas expander module separates gas bubbles from the liquid phase during transportation of the medium. Finally, a new type of gradient culture container allows a permanent elimination of transported gas bubbles. Application of this innovative equipment optimizes the parallel transportation of fluid in the luminal and basal compartments of a gradient culture container. (topical review)

Long-term outdoor cultivation by perfusing spent medium for biodiesel production from Chlorella minutissima.

Science.gov (United States)

Oh, Sung Ho; Kwon, Min Chul; Choi, Woon Yong; Seo, Yong Chang; Kim, Ga Bin; Kang, Do Hyung; Lee, Shin Young; Lee, Hyeon Yong

2010-08-01

A unique perfusion process was developed to maintain high concentrations of marine alga, Chlorella minutissima. This method is based on recycling cells by continuous feeding with warm spent sea water from nuclear power plants, which has very similar properties as sea water. A temperature of at least 30 degrees C in a 200 L photo-bioreactor was maintained in this system by perfusion of the thermal plume for 80 days in the coldest season. The maximum cell concentration and total lipid content was 8.3 g-dry wt./L and 23.2 %, w/w, respectively, under mixotrophic conditions. Lipid production was found to be due to a partially or non-growth related process, which implies that large amounts of biomass are needed for a high accumulation of lipids within the cells. At perfusion rates greater than 1.5 L/h, the temperature of the medium inside the reactor was around 30 degrees C, which was optimal for cell growth. For this system, a perfusion rate of 2.8 L/h was determined to be optimal for maintaining rapid cell growth and lipid production during outdoor cultivation. It was absolutely necessary to maintain the appropriate perfusion rate so that the medium temperature was optimal for cell growth. In addition, the lipids produced using this process were shown to be feasible for biodiesel production since the lipid composition of C. minutissima grown under these conditions consisted of 17 % (w/w) of C(16) and 47% (w/w) of C(18). The combined results of this study clearly demonstrated that the discharged energy of the thermal plume could be reused to cultivate marine alga by maintaining a relatively constant temperature in an outdoor photo-bioreactor without the need for supplying any extra energy, which could allow for cheap production of biodiesel from waste energy. Copyright 2010 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Scale up of diesel oil biodegradation in a baffled roller bioreactor.

Science.gov (United States)

Nikakhtari, Hossein; Song, Wanning; Kumar, Pardeep; Nemati, Mehdi; Hill, Gordon A

2010-05-01

Diesel oil is a suitable substance to represent petroleum contamination from accidental spills in operating and transportation facilities. Using a microbial culture enriched from a petroleum contaminated soil, biodegradation of diesel oil was carried out in 2.2, 55, and 220 L roller baffled bioreactors. The effects of bioreactor rotation speed (from 5 to 45 rpm) and liquid loading (from 18% to 73% of total volume) on the biodegradation of diesel oil were studied. In the small scale bioreactor (2.2L), the maximum rotation speed of 45 rpm resulted in the highest biodegradation rate with a first order biodegradation kinetic constant of 0.095 d(-1). In the larger scale bioreactors, rotation speed did not affect the biodegradation rate. Liquid loadings higher than 64% resulted in reduced biodegradation rates in the small scale bioreactor; however, in the larger roller bioreactors liquid loading did not affect the biodegradation rate. Biodegradation of diesel oil at 5 rpm and 73% loading is recommended for operating large scale roller baffled bioreactors. Under these conditions, high diesel oil concentrations up to 50 gL(-1) can be bioremediated at a rate of 1.61 gL(-1)d(-1). Copyright 2010 Elsevier Ltd. All rights reserved.

A fluid dynamics approach to bioreactor design for cell and tissue culture.

Science.gov (United States)

Dusting, Jonathan; Sheridan, John; Hourigan, Kerry

2006-08-20

The problem of controlling cylindrical tank bioreactor conditions for cell and tissue culture purposes has been considered from a flow dynamics perspective. Simple laminar flows in the vortex breakdown region are proposed as being a suitable alternative to turbulent spinner flask flows and horizontally oriented rotational flows. Vortex breakdown flows have been measured using three-dimensional Stereoscopic particle image velocimetry, and non-dimensionalized velocity and stress distributions are presented. Regions of locally high principal stress occur in the vicinity of the impeller and the lower sidewall. Topological changes in the vortex breakdown region caused by an increase in Reynolds number are reflected in a redistribution of the peak stress regions. The inclusion of submerged scaffold models adds complexity to the flow, although vortex breakdown may still occur. Relatively large stresses occur along the edge of disks jutting into the boundary of the vortex breakdown region. Copyright 2006 Wiley Periodicals, Inc.

A novel customizable modular bioreactor system for whole-heart cultivation under controlled 3D biomechanical stimulation.

Science.gov (United States)

Hülsmann, Jörn; Aubin, Hug; Kranz, Alexander; Godehardt, Erhardt; Munakata, Hiroshi; Kamiya, Hiroyuki; Barth, Mareike; Lichtenberg, Artur; Akhyari, Payam

2013-09-01

In the last decade, cardiovascular tissue engineering has made great progress developing new strategies for regenerative medicine applications. However, while tissue engineered heart valves are already entering the clinical routine, tissue engineered myocardial substitutes are still restrained to experimental approaches. In contrast to the heart valves, tissue engineered myocardium cannot be repopulated in vivo because of its biological complexity, requiring elaborate cultivation conditions ex vivo. Although new promising approaches-like the whole-heart decellularization concept-have entered the myocardial tissue engineering field, bioreactor technology needed for the generation of functional myocardial tissue still lags behind in the sense of user-friendly, flexible and low cost systems. Here, we present a novel customizable modular bioreactor system that can be used for whole-heart cultivation. Out of a commercially obtainable original equipment manufacturer platform we constructed a modular bioreactor system specifically aimed at the cultivation of decellularized whole-hearts through perfusion and controlled 3D biomechanical stimulation with a simple but highly flexible operation platform based on LabVIEW. The modular setup not only allows a wide range of variance regarding medium conditioning under controlled 3D myocardial stretching but can also easily be upgraded for e.g. electrophysiological monitoring or stimulation, allowing for a tailor-made low-cost myocardial bioreactor system.

The importance of bicarbonate and nonbicarbonate buffer systems in batch and continuous flow bioreactors for articular cartilage tissue engineering.

Science.gov (United States)

Khan, Aasma A; Surrao, Denver C

2012-05-01

In cartilage tissue engineering an optimized culture system, maintaining an appropriate extracellular environment (e.g., pH of media), can increase cell proliferation and extracellular matrix (ECM) accumulation. We have previously reported on a continuous-flow bioreactor that improves tissue growth by supplying the cells with a near infinite supply of medium. Previous studies have observed that acidic environments reduce ECM synthesis and chondrocyte proliferation. Hence, in this study we investigated the combined effects of a continuous culture system (bioreactor) together with additional buffering agents (e.g., sodium bicarbonate [NaHCO₃]) on cartilaginous tissue growth in vitro. Isolated bovine chondrocytes were grown in three-dimensional cultures, either in static conditions or in a continuous-flow bioreactor, in media with or without NaHCO₃. Tissue constructs cultivated in the bioreactor with NaHCO₃-supplemented media were characterized with significantly increased (p<0.05) ECM accumulation (glycosaminoglycans a 98-fold increase; collagen a 25-fold increase) and a 13-fold increase in cell proliferation, in comparison with static cultures. Additionally, constructs grown in the bioreactor with NaHCO₃-supplemented media were significantly thicker than all other constructs (p<0.05). Further, the chondrocytes from the primary construct expanded and synthesized ECM, forming a secondary construct without a separate expansion phase, with a diameter and thickness of 4 mm and 0.72 mm respectively. Tissue outgrowth was negligible in all other culturing conditions. Thus this study demonstrates the advantage of employing a continuous flow bioreactor coupled with NaHCO₃ supplemented media for articular cartilage tissue engineering.

A perfusion culture system using a stirred ceramic membrane reactor for hyperproduction of IgG2a monoclonal antibody by hybridoma cells.

Science.gov (United States)

Dong, Haodi; Tang, Ya-Jie; Ohashi, Ryo; Hamel, Jean-François P

2005-01-01

A novel perfusion culture system for efficient production of IgG2a monoclonal antibody (mAb) by hybridoma cells was developed. A ceramic membrane module was constructed and used as a cell retention device installed in a conventional stirred-tank reactor during the perfusion culture. Furthermore, the significance of the control strategy of perfusion rate (volume of fresh medium/working volume of reactor/day, vvd) was investigated. With the highest increasing rate (deltaD, vvd per day, vvdd) of perfusion rate, the maximal viable cell density of 3.5 x 10(7) cells/mL was obtained within 6 days without any limitation and the cell viability was maintained above 95%. At lower deltaD's, the cell growth became limited. Under nutrient-limited condition, the specific cell growth rate (mu) was regulated by deltaD. During the nonlimited growth phase, the specific mAb production rate (qmAb) remained constant at 0.26 +/- 0.02 pg/cell x h in all runs. During the cell growth-limited phase, qmAb was regulated by deltaD within the range of 0.25-0.65 vvdd. Under optimal conditions, qmAb of 0.80 and 2.15 pg/cell x h was obtained during the growth-limited phase and stationary phase, respectively. The overall productivity and yield were 690 mg/L x day and 340 mg/L x medium, respectively. This study demonstrated that this novel perfusion culture system for suspension mammalian cells can support high cell density and efficient mAb production and that deltaD is an important control parameter to regulate and achieve high mAb production.

On-line monitoring of monoclonal antibody formation in high density perfusion culture using FIA.

Science.gov (United States)

Fenge, C; Fraune, E; Freitag, R; Scheper, T; Schügerl, K

1991-05-01

An automated flow injection system for on-line analysis of proteins in real fermentation fluids was developed by combining the principles of stopped-flow, merging zones flow injection analysis (FIA) with antigen-antibody reactions. IgG in the sample reacted with its corresponding antibody (a-IgG) in the reagent solution. Formation of insoluble immunocomplexes resulted in an increase of the turbidity which was determined photometrically. This system was used to monitor monoclonal antibody production in high cell density perfusion culture of hybridoma cells. Perfusion was performed with a newly developed static filtration unit equipped with hydrophilic microporous tubular membranes. Different sampling devices were tested to obtain a cell-free sample stream for on-line product analysis of high molecular weight (e.g., monoclonal antibodies) and low molecular weight (e.g., glucose, lactate) medium components. In fermentation fluids a good correlation (coefficient: 0.996) between the FIA method and an ELISA test was demonstrated. In a high density perfusion cultivation process mAb formation was successfully monitored on-line over a period of 400 h using a reliable sampling system. Glucose and lactate were measured over the same period of time using a commercially available automatic analyser based on immobilized enzyme technology.

Design considerations and challenges for mechanical stretch bioreactors in tissue engineering.

Science.gov (United States)

Lei, Ying; Ferdous, Zannatul

2016-05-01

With the increase in average life expectancy and growing aging population, lack of functional grafts for replacement surgeries has become a severe problem. Engineered tissues are a promising alternative to this problem because they can mimic the physiological function of the native tissues and be cultured on demand. Cyclic stretch is important for developing many engineered tissues such as hearts, heart valves, muscles, and bones. Thus a variety of stretch bioreactors and corresponding scaffolds have been designed and tested to study the underlying mechanism of tissue formation and to optimize the mechanical conditions applied to the engineered tissues. In this review, we look at various designs of stretch bioreactors and common scaffolds and offer insights for future improvements in tissue engineering applications. First, we summarize the requirements and common configuration of stretch bioreactors. Next, we present the features of different actuating and motion transforming systems and their applications. Since most bioreactors must measure detailed distributions of loads and deformations on engineered tissues, techniques with high accuracy, precision, and frequency have been developed. We also cover the key points in designing culture chambers, nutrition exchanging systems, and regimens used for specific tissues. Since scaffolds are essential for providing biophysical microenvironments for residing cells, we discuss materials and technologies used in fabricating scaffolds to mimic anisotropic native tissues, including decellularized tissues, hydrogels, biocompatible polymers, electrospinning, and 3D bioprinting techniques. Finally, we present the potential future directions for improving stretch bioreactors and scaffolds. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:543-553, 2016. © 2016 American Institute of Chemical Engineers.

Towards a Tissue-Engineered Ligament: Design and Preliminary Evaluation of a Dedicated Multi-Chamber Tension-Torsion Bioreactor

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Cédric P. Laurent

2014-02-01

Full Text Available Tissue engineering may constitute a promising alternative to current strategies in ligament repair, providing that suitable scaffolds and culture conditions are proposed. The objective of the present contribution is to present the design and instrumentation of a novel multi-chamber tension-torsion bioreactor dedicated to ligament tissue engineering. A preliminary biological evaluation of a new braided scaffold within this bioreactor under dynamic loading is reported, starting with the development of a dedicated seeding protocol validated from static cultures. The results of these preliminary biological characterizations confirm that the present combination of scaffold, seeding protocol and bioreactor may enable us to head towards a suitable ligament tissue-engineered construct.

Bioreactors as Engineering Support to Treat Cardiac Muscle and Vascular Disease

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Diana Massai

2013-01-01

Full Text Available Cardiovascular disease is the leading cause of morbidity and mortality in the Western World. The inability of fully differentiated, load-bearing cardiovascular tissues to in vivo regenerate and the limitations of the current treatment therapies greatly motivate the efforts of cardiovascular tissue engineering to become an effective clinical strategy for injured heart and vessels. For the effective production of organized and functional cardiovascular engineered constructs in vitro, a suitable dynamic environment is essential, and can be achieved and maintained within bioreactors. Bioreactors are technological devices that, while monitoring and controlling the culture environment and stimulating the construct, attempt to mimic the physiological milieu. In this study, a review of the current state of the art of bioreactor solutions for cardiovascular tissue engineering is presented, with emphasis on bioreactors and biophysical stimuli adopted for investigating the mechanisms influencing cardiovascular tissue development, and for eventually generating suitable cardiovascular tissue replacements.

Characterizing steady states of genome-scale metabolic networks in continuous cell cultures.

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Jorge Fernandez-de-Cossio-Diaz

2017-11-01

Full Text Available In the continuous mode of cell culture, a constant flow carrying fresh media replaces culture fluid, cells, nutrients and secreted metabolites. Here we present a model for continuous cell culture coupling intra-cellular metabolism to extracellular variables describing the state of the bioreactor, taking into account the growth capacity of the cell and the impact of toxic byproduct accumulation. We provide a method to determine the steady states of this system that is tractable for metabolic networks of arbitrary complexity. We demonstrate our approach in a toy model first, and then in a genome-scale metabolic network of the Chinese hamster ovary cell line, obtaining results that are in qualitative agreement with experimental observations. We derive a number of consequences from the model that are independent of parameter values. The ratio between cell density and dilution rate is an ideal control parameter to fix a steady state with desired metabolic properties. This conclusion is robust even in the presence of multi-stability, which is explained in our model by a negative feedback loop due to toxic byproduct accumulation. A complex landscape of steady states emerges from our simulations, including multiple metabolic switches, which also explain why cell-line and media benchmarks carried out in batch culture cannot be extrapolated to perfusion. On the other hand, we predict invariance laws between continuous cell cultures with different parameters. A practical consequence is that the chemostat is an ideal experimental model for large-scale high-density perfusion cultures, where the complex landscape of metabolic transitions is faithfully reproduced.

Differentiation of mammalian skeletal muscle cells cultured on microcarrier beads in a rotating cell culture system

Science.gov (United States)

Torgan, C. E.; Burge, S. S.; Collinsworth, A. M.; Truskey, G. A.; Kraus, W. E.

2000-01-01

The growth and repair of adult skeletal muscle are due in part to activation of muscle precursor cells, commonly known as satellite cells or myoblasts. These cells are responsive to a variety of environmental cues, including mechanical stimuli. The overall goal of the research is to examine the role of mechanical signalling mechanisms in muscle growth and plasticity through utilisation of cell culture systems where other potential signalling pathways (i.e. chemical and electrical stimuli) are controlled. To explore the effects of decreased mechanical loading on muscle differentiation, mammalian myoblasts are cultured in a bioreactor (rotating cell culture system), a model that has been utilised to simulate microgravity. C2C12 murine myoblasts are cultured on microcarrier beads in a bioreactor and followed throughout differentiation as they form a network of multinucleated myotubes. In comparison with three-dimensional control cultures that consist of myoblasts cultured on microcarrier beads in teflon bags, myoblasts cultured in the bioreactor exhibit an attenuation in differentiation. This is demonstrated by reduced immunohistochemical staining for myogenin and alpha-actinin. Western analysis shows a decrease, in bioreactor cultures compared with control cultures, in levels of the contractile proteins myosin (47% decrease, p < 0.01) and tropomyosin (63% decrease, p < 0.01). Hydrodynamic measurements indicate that the decrease in differentiation may be due, at least in part, to fluid stresses acting on the myotubes. In addition, constraints on aggregate size imposed by the action of fluid forces in the bioreactor affect differentiation. These results may have implications for muscle growth and repair during spaceflight.

Effect of changing temperature on anaerobic hydrogen production and microbial community composition in an open-mixed culture bioreactor

Energy Technology Data Exchange (ETDEWEB)

Karadag, Dogan; Puhakka, Jaakko A. [Department of Chemistry and Bioengineering, Tampere University of Technology, Tampere (Finland)

2010-10-15

The temperature effect (37-65 C) on H{sub 2} production from glucose in an open-mixed culture bioreactor using an enrichment culture from a hot spring was studied. The dynamics of microbial communities was investigated by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). At 45 and 60 C the H{sub 2} production was the highest i.e. 1.71 and 0.85 mol H{sub 2}/mol glucose, respectively. No H{sub 2} was produced at temperatures 50 and 55 C. At 37-45 C, H{sub 2} production was produced by butyrate type fermentation while fermentation mechanism changed to ethanol type at 60 C. Clostridium species were dominant at 37-45 C while at 50-55 C and 60 C the culture was dominated by Bacillus coagulans and Thermoanaerobacterium, respectively. In the presence of B. Coagulans the metabolism was directed to lactate production. The results show that the mixed culture had two optima for H{sub 2} production and that the microbial communities and metabolic patterns promptly changed according to changing temperatures. (author)

Catalytic bioreactors and methods of using same

Science.gov (United States)

Worden, Robert Mark; Liu, Yangmu Chloe

2017-07-25

Various embodiments provide a bioreactor for producing a bioproduct comprising one or more catalytically active zones located in a housing and adapted to keep two incompatible gaseous reactants separated when in a gas phase, wherein each of the one or more catalytically active zones may comprise a catalytic component retainer and a catalytic component retained within and/or thereon. Each of the catalytically active zones may additionally or alternatively comprise a liquid medium located on either side of the catalytic component retainer. Catalytic component may include a microbial cell culture located within and/or on the catalytic component retainer, a suspended catalytic component suspended in the liquid medium, or a combination thereof. Methods of using various embodiments of the bioreactor to produce a bioproduct, such as isobutanol, are also provided.

Liver tissue engineering based on aggregate assembly: efficient formation of endothelialized rat hepatocyte aggregates and their immobilization with biodegradable fibres

International Nuclear Information System (INIS)

Pang, Y; Shinohara, M; Komori, K; Sakai, Y; Montagne, K

2012-01-01

To realize long-term in vitro culture of hepatocytes at a high density while maintaining a high hepatic function for aggregate-based liver tissue engineering, we report here a novel culture method whereby endothelialized rat hepatocyte aggregates were formed using a PDMS microwell device and cultured in a perfusion bioreactor by introducing spacers between aggregates to improve oxygen and nutrient supply. Primary rat hepatocyte aggregates around 100 µm in diameter coated with human umbilical vein endothelial cells were spontaneously and quickly formed after 12 h of incubation, thanks to the continuous supply of oxygen by diffusion through the PDMS honeycomb microwell device. Then, the recovered endothelialized rat hepatocyte aggregates were mixed with biodegradable poly-l-lactic acid fibres in suspension and packed into a PDMS-based bioreactor. Perfusion culture of 7 days was successfully achieved with more than 73.8% cells retained in the bioreactor. As expected, the fibres acted as spacers between aggregates, which was evidenced from the enhanced albumin production and more spherical morphology compared with fibre-free packing. In summary, this study shows the advantages of using PDMS-based microwells to form heterotypic aggregates and also demonstrates the feasibility of spacing tissue elements for improving oxygen and nutrient supply to tissue engineering based on modular assembly. (paper)

Bacterial community dynamics during start-up of a trickle-bed bioreactor degrading aromatic compounds.

Science.gov (United States)

Stoffels, M; Amann, R; Ludwig, W; Hekmat, D; Schleifer, K H

1998-03-01

This study was performed with a laboratory-scale fixed-bed bioreactor degrading a mixture of aromatic compounds (Solvesso100). The starter culture for the bioreactor was prepared in a fermentor with a wastewater sample of a care painting facility as the inoculum and Solvesso100 as the sole carbon source. The bacterial community dynamics in the fermentor and the bioreactor were examined by a conventional isolation procedure and in situ hybridization with fluorescently labeled rRNA-targeted oligonucleotides. Two significant shifts in the bacterial community structure could be demonstrated. The original inoculum from the wastewater of the car factory was rich in proteobacteria of the alpha and beta subclasses, while the final fermentor enrichment was dominated by bacteria closely related to Pseudomonas putida or Pseudomonas mendocina, which both belong to the gamma subclass of the class Proteobacteria. A second significant shift was observed when the fermentor culture was transferred as inoculum to the trickle-bed bioreactor. The community structure in the bioreactor gradually returned to a higher complexity, with the dominance of beta and alpha subclass proteobacteria, whereas the gamma subclass proteobacteria sharply declined. Obviously, the preceded pollutant adaptant did not lead to a significant enrichment of bacteria that finally dominated in the trickle-bed bioreactor. In the course of experiments, three new 16S as well as 23S rRNA-targeted probes for beta subclass proteobacteria were designed, probe SUBU1237 for the genera Burkholderia and Sutterella, probe ALBO34a for the genera Alcaligenes and Bordetella, and probe Bcv13b for Burkholderia cepacia and Burkholderia vietnamiensis. Bacteria hybridizing with the probe Bcv13b represented the main Solvesso100-degrading population in the reactor.

Bioreactors for Tissue Engineering of Cartilage

Science.gov (United States)

Concaro, S.; Gustavson, F.; Gatenholm, P.

The cartilage regenerative medicine field has evolved during the last decades. The first-generation technology, autologous chondrocyte transplantation (ACT) involved the transplantation of in vitro expanded chondrocytes to cartilage defects. The second generation involves the seeding of chondrocytes in a three-dimensional scaffold. The technique has several potential advantages such as the ability of arthroscopic implantation, in vitro pre-differentiation of cells and implant stability among others (Brittberg M, Lindahl A, Nilsson A, Ohlsson C, Isaksson O, Peterson L, N Engl J Med 331(14):889-895, 1994; Henderson I, Francisco R, Oakes B, Cameron J, Knee 12(3):209-216, 2005; Peterson L, Minas T, Brittberg M, Nilsson A, Sjogren-Jansson E, Lindahl A, Clin Orthop (374):212-234, 2000; Nagel-Heyer S, Goepfert C, Feyerabend F, Petersen JP, Adamietz P, Meenen NM, et al. Bioprocess Biosyst Eng 27(4):273-280, 2005; Portner R, Nagel-Heyer S, Goepfert C, Adamietz P, Meenen NM, J Biosci Bioeng 100(3):235-245, 2005; Nagel-Heyer S, Goepfert C, Adamietz P, Meenen NM, Portner R, J Biotechnol 121(4):486-497, 2006; Heyland J, Wiegandt K, Goepfert C, Nagel-Heyer S, Ilinich E, Schumacher U, et al. Biotechnol Lett 28(20):1641-1648, 2006). The nutritional requirements of cells that are synthesizing extra-cellular matrix increase along the differentiation process. The mass transfer must be increased according to the tissue properties. Bioreactors represent an attractive tool to accelerate the biochemical and mechanical properties of the engineered tissues providing adequate mass transfer and physical stimuli. Different reactor systems have been [5] developed during the last decades based on different physical stimulation concepts. Static and dynamic compression, confined and nonconfined compression-based reactors have been described in this review. Perfusion systems represent an attractive way of culturing constructs under dynamic conditions. Several groups showed increased matrix

Production of diosgenin from Dioscorea zingiberensis with mixed culture in a new tray bioreactor

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Yutong Cheng

2016-01-01

Full Text Available A new tray bioreactor was developed for the production of diosgenin from Dioscorea zingiberensis with Trichoderma reesei and Aspergillus fumigatus. The influence of initial moisture content, temperature, tray bed depth and mixing times was investigated. The best fermentation condition is initial moisture content of 75%, bioreactor temperature of 35°C, solid bed depth of 1.5 cm and three mixings carrying out on the first, third and fifth day. Under the optimized fermentation conditions, after 144 h incubation, maximum diogenin concentration of 68.2 μmol/g was detected.

Solvent Fermentation From Palm Oil Mill Effluent Using Clostridium acetobutylicum In Oscillatory Flow Bioreactor

International Nuclear Information System (INIS)

Takriff, M.S.; Masngut, N.; Kadhum, A.A.H.; Kalil, M.S.; Mohammad, A.W.

2009-01-01

Acetone-butanol-ethanol (ABE) fermentation from Palm Oil Mill Effluent (POME) by C. acetobutylicum NCIMB 13357 in an oscillatory flow bioreactor was investigated. Experimental works were conducted in a U-shaped stainless steel oscillatory flow bioreactor at oscillation frequency between 0.45-0.78 Hz and a constant amplitude of 12.5 mm. Fermentations were carried out for 72 hr at 35 degree Celsius using palm oil mill effluent and reinforced clostridia medium as a growth medium in batch culture. Result of this investigation showed that POME is a viable media for ABE fermentation and oscillatory flow bioreactor has an excellent potential as an alternative fermentation device. (author)

High cell density suppresses BMP4-induced differentiation of human pluripotent stem cells to produce macroscopic spatial patterning in a unidirectional perfusion culture chamber.

Science.gov (United States)

Tashiro, Shota; Le, Minh Nguyen Tuyet; Kusama, Yuta; Nakatani, Eri; Suga, Mika; Furue, Miho K; Satoh, Taku; Sugiura, Shinji; Kanamori, Toshiyuki; Ohnuma, Kiyoshi

2018-04-19

Spatial pattern formation is a critical step in embryogenesis. Bone morphogenetic protein 4 (BMP4) and its inhibitors are major factors for the formation of spatial patterns during embryogenesis. However, spatial patterning of the human embryo is unclear because of ethical issues and isotropic culture environments resulting from conventional culture dishes. Here, we utilized human pluripotent stem cells (hiPSCs) and a simple anisotropic (unidirectional perfusion) culture chamber, which creates unidirectional conditions, to measure the cell community effect. The influence of cell density on BMP4-induced differentiation was explored during static culture using a conventional culture dish. Immunostaining of the early differentiation marker SSEA-1 and the mesendoderm marker BRACHYURY revealed that high cell density suppressed differentiation, with small clusters of differentiated and undifferentiated cells formed. Addition of five-fold higher concentration of BMP4 showed similar results, suggesting that suppression was not caused by depletion of BMP4 but rather by high cell density. Quantitative RT-PCR array analysis showed that BMP4 induced multi-lineage differentiation, which was also suppressed under high-density conditions. We fabricated an elongated perfusion culture chamber, in which proteins were transported unidirectionally, and hiPSCs were cultured with BMP4. At low density, the expression was the same throughout the chamber. However, at high density, SSEA-1 and BRACHYURY were expressed only in upstream cells, suggesting that some autocrine/paracrine factors inhibited the action of BMP4 in downstream cells to form the spatial pattern. Human iPSCs cultured in a perfusion culture chamber might be useful for studying in vitro macroscopic pattern formation in human embryogenesis. Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Farm Deployable Microbial Bioreactor for Fuel Ethanol Production

Energy Technology Data Exchange (ETDEWEB)

Okeke, Benedict [Auburn Univ., Montgomery AL (United States)

2016-03-30

Research was conducted to develop a farm and field deployable microbial bioreactor for bioethanol production from biomass. Experiments were conducted to select the most efficient microorganisms for conversion of plant fiber to sugars for fermentation to ethanol. Mixtures of biomass and surface soil samples were collected from selected sites in Alabama black belt counties (Macon, Sumter, Choctaw, Dallas, Montgomery, Lowndes) and other areas within the state of Alabama. Experiments were conducted to determine the effects of culture parameters on key biomass saccharifying enzymes (cellulase, beta-glucosidase, xylanase and beta-xylosidase). A wide-scale sampling of locally-grown fruits in Central Alabama was embarked to isolate potential xylose fermenting microorganisms. Yeast isolates were evaluated for xylose fermentation. Selected microorganisms were characterized by DNA based methods. Factors affecting enzyme production and biomass saccharification were examined and optimized in the laboratory. Methods of biomass pretreatment were compared. Co-production of amylolytic enzymes with celluloytic-xylanolytic enzymes was evaluated; and co-saccharification of a combination of biomass, and starch-rich materials was examined. Simultaneous saccharification and fermentation with and without pre-saccharifcation was studied. Whole culture broth and filtered culture broth simultaneous saccahrifcation and fermentation were compared. A bioreactor system was designed and constructed to employ laboratory results for scale up of biomass saccharification.

Assessing the culture of safety in cardiovascular perfusion: attitudes and perceptions.

Science.gov (United States)

Lawson, Chad; Predella, Megan; Rowden, Allison; Goldstein, Jamie; Sistino, Joseph J; Fitzgerald, David C

2017-10-01

The Hospital Survey on Patient Safety Culture was developed by the Agency for Healthcare Research and Quality (AHRQ) to assess the culture of safety in hospitals. The purpose of this study was to identify specific domains of perfusion that are indicators of a high quality culture of safety. Perfusionists were recruited to participate in the survey through email invitation through Perflist, Perfmail and LinkedIn. The survey consisted of 37 questions across six safety domains. Questions were developed using the AHRQ Hospital Survey on Patient Safety Culture. 'Positive scores' were defined as a response that either agreed or strongly agreed with a safety standard. Survey responses that resulted in a 75 percent or higher positive response rate were identified as vital components of a high culture of safety. Logistic regression analysis was used to determine importance components of perceived safety. Four responses were found to have a significant predictive level of a positive safety environment in the work unit: (1) in this unit, we discuss ways to prevent errors from happening again; OR=3.09, (2) in this unit, we treat others with respect; OR=1.09 (3) my supervisor/manager seriously considers staff suggestions for improving patient safety; OR=1.89 and (4) there is good cooperation among hospital units that need to work together; OR=1.77. There were two predictors of a negative work unit safety environment: (1) staff are afraid to ask questions when something does not seem right; OR=0.62 and (2) it is just by chance that more serious mistakes don't happen around here; OR=0.55. The results from this survey indicate that effective communication secondary to both incident and near-miss reporting is associated with a higher perceived culture of safety. A positive safety environment is associated with being able to speak up regarding safety issues without fear of negative repercussions.

Bioreactors for plant cells: hardware configuration and internal environment optimization as tools for wider commercialization.

Science.gov (United States)

Georgiev, Milen I; Weber, Jost

2014-07-01

Mass production of value-added molecules (including native and heterologous therapeutic proteins and enzymes) by plant cell culture has been demonstrated as an efficient alternative to classical technologies [i.e. natural harvest and chemical (semi)synthesis]. Numerous proof-of-concept studies have demonstrated the feasibility of scaling up plant cell culture-based processes (most notably to produce paclitaxel) and several commercial processes have been established so far. The choice of a suitable bioreactor design (or modification of an existing commercially available reactor) and the optimization of its internal environment have been proven as powerful tools toward successful mass production of desired molecules. This review highlights recent progress (mostly in the last 5 years) in hardware configuration and optimization of bioreactor culture conditions for suspended plant cells.

Design and Validation of a Cyclic Strain Bioreactor to Condition Spatially-Selective Scaffolds in Dual Strain Regimes

Directory of Open Access Journals (Sweden)

J. Matthew Goodhart

2014-03-01

Full Text Available The objective of this study was to design and validate a unique bioreactor design for applying spatially selective, linear, cyclic strain to degradable and non-degradable polymeric fabric scaffolds. This system uses a novel three-clamp design to apply cyclic strain via a computer controlled linear actuator to a specified zone of a scaffold while isolating the remainder of the scaffold from strain. Image analysis of polyethylene terephthalate (PET woven scaffolds subjected to a 3% mechanical stretch demonstrated that the stretched portion of the scaffold experienced 2.97% ± 0.13% strain (mean ± standard deviation while the unstretched portion experienced 0.02% ± 0.18% strain. NIH-3T3 fibroblast cells were cultured on the PET scaffolds and half of each scaffold was stretched 5% at 0.5 Hz for one hour per day for 14 days in the bioreactor. Cells were checked for viability and proliferation at the end of the 14 day period and levels of glycosaminoglycan (GAG and collagen (hydroxyproline were measured as indicators of extracellular matrix production. Scaffolds in the bioreactor showed a seven-fold increase in cell number over scaffolds cultured statically in tissue culture plastic petri dishes (control. Bioreactor scaffolds showed a lower concentration of GAG deposition per cell as compared to the control scaffolds largely due to the great increase in cell number. A 75% increase in hydroxyproline concentration per cell was seen in the bioreactor stretched scaffolds as compared to the control scaffolds. Surprisingly, little differences were experienced between the stretched and unstretched portions of the scaffolds for this study. This was largely attributed to the conditioned and shared media effect. Results indicate that the bioreactor system is capable of applying spatially-selective, linear, cyclic strain to cells growing on polymeric fabric scaffolds and evaluating the cellular and matrix responses to the applied strains.

AMC-Bio-Artificial Liver culturing enhances mitochondrial biogenesis in human liver cell lines: The role of oxygen, medium perfusion and 3D configuration

NARCIS (Netherlands)

Adam, Aziza A. A.; van Wenum, Martien; van der Mark, Vincent A.; Jongejan, Aldo; Moerland, Perry D.; Houtkooper, Riekelt H.; Wanders, Ronald J. A.; Oude Elferink, Ronald P.; Chamuleau, Robert A. F. M.; Hoekstra, Ruurdtje

2017-01-01

Human liver cell lines, like HepaRG and C3A, acquire higher functionality when cultured in the AMC-Bio-Artificial Liver (AMC-BAL). The three main differences between BAL and monolayer culture are the oxygenation (40% vs 20%O2), dynamic vs absent medium perfusion and 3D vs 2D configuration. Here, we

Advances in biotreatment of acid mine drainage and biorecovery of metals: 2. Membrane bioreactor system for sulfate reduction.

Science.gov (United States)

Tabak, Henry H; Govind, Rakesh

2003-12-01

Several biotreatmemt techniques for sulfate conversion by the sulfate reducing bacteria (SRB) have been proposed in the past, however few of them have been practically applied to treat sulfate containing acid mine drainage (AMD). This research deals with development of an innovative polypropylene hollow fiber membrane bioreactor system for the treatment of acid mine water from the Berkeley Pit, Butte, MT, using hydrogen consuming SRB biofilms. The advantages of using the membrane bioreactor over the conventional tall liquid phase sparged gas bioreactor systems are: large microporous membrane surface to the liquid phase; formation of hydrogen sulfide outside the membrane, preventing the mixing with the pressurized hydrogen gas inside the membrane; no requirement of gas recycle compressor; membrane surface is suitable for immobilization of active SRB, resulting in the formation of biofilms, thus preventing washout problems associated with suspended culture reactors; and lower operating costs in membrane bioreactors, eliminating gas recompression and gas recycle costs. Information is provided on sulfate reduction rate studies and on biokinetic tests with suspended SRB in anaerobic digester sludge and sediment master culture reactors and with SRB biofilms in bench-scale SRB membrane bioreactors. Biokinetic parameters have been determined using biokinetic models for the master culture and membrane bioreactor systems. Data are presented on the effect of acid mine water sulfate loading at 25, 50, 75 and 100 ml/min in scale-up SRB membrane units, under varied temperatures (25, 35 and 40 degrees C) to determine and optimize sulfate conversions for an effective AMD biotreatment. Pilot-scale studies have generated data on the effect of flow rates of acid mine water (MGD) and varied inlet sulfate concentrations in the influents on the resultant outlet sulfate concentration in the effluents and on the number of SRB membrane modules needed for the desired sulfate conversion in

Animal cell cultures: recent achievements and perspectives in the production of biopharmaceuticals.

Science.gov (United States)

Butler, Michael

2005-08-01

There has been a rapid increase in the number and demand for approved biopharmaceuticals produced from animal cell culture processes over the last few years. In part, this has been due to the efficacy of several humanized monoclonal antibodies that are required at large doses for therapeutic use. There have also been several identifiable advances in animal cell technology that has enabled efficient biomanufacture of these products. Gene vector systems allow high specific protein expression and some minimize the undesirable process of gene silencing that may occur in prolonged culture. Characterization of cellular metabolism and physiology has enabled the design of fed-batch and perfusion bioreactor processes that has allowed a significant improvement in product yield, some of which are now approaching 5 g/L. Many of these processes are now being designed in serum-free and animal-component-free media to ensure that products are not contaminated with the adventitious agents found in bovine serum. There are several areas that can be identified that could lead to further improvement in cell culture systems. This includes the down-regulation of apoptosis to enable prolonged cell survival under potentially adverse conditions. The characterization of the critical parameters of glycosylation should enable process control to reduce the heterogeneity of glycoforms so that production processes are consistent. Further improvement may also be made by the identification of glycoforms with enhanced biological activity to enhance clinical efficacy. The ability to produce the ever-increasing number of biopharmaceuticals by animal cell culture is dependent on sufficient bioreactor capacity in the industry. A recent shortfall in available worldwide culture capacity has encouraged commercial activity in contract manufacturing operations. However, some analysts indicate that this still may not be enough and that future manufacturing demand may exceed production capacity as the number

Tubular bioreactor and its application; Tubular bioreactor to sono tekiyo

Energy Technology Data Exchange (ETDEWEB)

Endo, I.; Nagamune, T. [The University of Tokyo, Tokyo (Japan). Faculty of Engineering; Yuki, K. [Nikka Whisky Distilling Co. Ltd. Tokyo (Japan); Inaba, H. [Sumitomo Heavy Industries, Ltd., Tokyo (Japan)

1994-09-05

The loop type tubular bioreactor (TBR) was developed where biocatalysts are trapped in the reactor by membrane module. A UF membrane or MF membrane and crossflow filtration were adopted for the membrane module, and the reactor loop was composed of four membrane modules. The reactor was operated at 2-4 m/s in membrane surface velocity and 300-400 kPa in filtration pressure. As the result of the high-density culture of lactic acid bacteria and yeast, a biomass concentration was more than 10 times that in batch culture, suggesting the remarkable enhancement of a production efficiency. As the result of the continuous fermentation of cider, the fast fermentation more than 60 times that in conventional ones was obtained together with the same quality as conventional ones. Such a fast fermentation was probably achieved by yeast suspended in the fermenter of TBR, by yeast hardly affected physico-chemically as compared with immobilized reactors, and by small effect of mass transfer on reaction systems. 4 refs., 6 figs.

Evaluation of the growth environment of a hydrostatic force bioreactor for preconditioning of tissue-engineered constructs.

Science.gov (United States)

Reinwald, Yvonne; Leonard, Katherine H L; Henstock, James R; Whiteley, Jonathan P; Osborne, James M; Waters, Sarah L; Levesque, Philippe; El Haj, Alicia J

2015-01-01

Bioreactors have been widely acknowledged as valuable tools to provide a growth environment for engineering tissues and to investigate the effect of physical forces on cells and cell-scaffold constructs. However, evaluation of the bioreactor environment during culture is critical to defining outcomes. In this study, the performance of a hydrostatic force bioreactor was examined by experimental measurements of changes in dissolved oxygen (O2), carbon dioxide (CO2), and pH after mechanical stimulation and the determination of physical forces (pressure and stress) in the bioreactor through mathematical modeling and numerical simulation. To determine the effect of hydrostatic pressure on bone formation, chick femur skeletal cell-seeded hydrogels were subjected to cyclic hydrostatic pressure at 0-270 kPa and 1 Hz for 1 h daily (5 days per week) over a period of 14 days. At the start of mechanical stimulation, dissolved O2 and CO2 in the medium increased and the pH of the medium decreased, but remained within human physiological ranges. Changes in physiological parameters (O2, CO2, and pH) were reversible when medium samples were placed in a standard cell culture incubator. In addition, computational modeling showed that the distribution and magnitude of physical forces depends on the shape and position of the cell-hydrogel constructs in the tissue culture format. Finally, hydrostatic pressure was seen to enhance mineralization of chick femur skeletal cell-seeded hydrogels.

Solid substrate fermentation of lignite by the coal-solubilizing mould, Trichoderma atroviride, in a new type of bioreactor

Energy Technology Data Exchange (ETDEWEB)

Holker, U.; Hofer, M. [University of Bonn, Bonn (Germany)

2002-07-01

Trichoderma atroviride CBS 349 is able to solubilize lignite. The mould was cultured under non-sterile conditions in a new type of bioreactor for solid substrate fermentation. German lignite (lithotype A, Bergheim) was used as complex solid substrate. Over 40 days 140 g of 1.5 kg lignite held in a 25 1-bioreactor was solubilized by the fungus.

Bioprocess development for kefiran production by Lactobacillus kefiranofaciens in semi industrial scale bioreactor

Directory of Open Access Journals (Sweden)

Daniel Joe Dailin

2016-07-01

Full Text Available Lactobacillus kefiranofaciens is non-pathogenic gram positive bacteria isolated from kefir grains and able to produce extracellular exopolysaccharides named kefiran. This polysaccharide contains approximately equal amounts of glucose and galactose. Kefiran has wide applications in pharmaceutical industries. Therefore, an approach has been extensively studied to increase kefiran production for pharmaceutical application in industrial scale. The present work aims to maximize kefiran production through the optimization of medium composition and production in semi industrial scale bioreactor. The composition of the optimal medium for kefiran production contained sucrose, yeast extract and K2HPO4 at 20.0, 6.0, 0.25 g L−1, respectively. The optimized medium significantly increased both cell growth and kefiran production by about 170.56% and 58.02%, respectively, in comparison with the unoptimized medium. Furthermore, the kinetics of cell growth and kefiran production in batch culture of L. kefiranofaciens was investigated under un-controlled pH conditions in 16-L scale bioreactor. The maximal cell mass in bioreactor culture reached 2.76 g L−1 concomitant with kefiran production of 1.91 g L−1.

Bioprocess development for kefiran production by Lactobacillus kefiranofaciens in semi industrial scale bioreactor.

Science.gov (United States)

Dailin, Daniel Joe; Elsayed, Elsayed Ahmed; Othman, Nor Zalina; Malek, Roslinda; Phin, Hiew Siaw; Aziz, Ramlan; Wadaan, Mohamad; El Enshasy, Hesham Ali

2016-07-01

Lactobacillus kefiranofaciens is non-pathogenic gram positive bacteria isolated from kefir grains and able to produce extracellular exopolysaccharides named kefiran. This polysaccharide contains approximately equal amounts of glucose and galactose. Kefiran has wide applications in pharmaceutical industries. Therefore, an approach has been extensively studied to increase kefiran production for pharmaceutical application in industrial scale. The present work aims to maximize kefiran production through the optimization of medium composition and production in semi industrial scale bioreactor. The composition of the optimal medium for kefiran production contained sucrose, yeast extract and K2HPO4 at 20.0, 6.0, 0.25 g L(-1), respectively. The optimized medium significantly increased both cell growth and kefiran production by about 170.56% and 58.02%, respectively, in comparison with the unoptimized medium. Furthermore, the kinetics of cell growth and kefiran production in batch culture of L. kefiranofaciens was investigated under un-controlled pH conditions in 16-L scale bioreactor. The maximal cell mass in bioreactor culture reached 2.76 g L(-1) concomitant with kefiran production of 1.91 g L(-1).

Binding and Conversion of Selenium in Candida utilis ATCC 9950 Yeasts in Bioreactor Culture

Directory of Open Access Journals (Sweden)

Marek Kieliszek

2017-02-01

Full Text Available Selenium is considered an essential component of all living organisms. The use of yeasts as a selenium supplement in human nutrition has gained much interest over the last decade. The accumulation and biochemical transformation of selenium in yeast cells is particularly interesting to many researchers. In this article, we present the results of the determination of selenium and selenomethionine content in the biomass of feed yeast Candida utilis ATCC 9950 obtained from the culture grown in a bioreactor. The results indicated that C. utilis cells performed the biotransformation of inorganic selenium(IV to organic derivatives (e.g., selenomethionine. Selenium introduced (20–30 mg Se4+∙L−1 to the experimental media in the form of sodium(IV selenite (Na2SeO3 salt caused a significant increase in selenium content in the biomass of C. utilis,irrespective of the concentration. The highest amount of selenium (1841 μg∙gd.w.−1 was obtained after a 48-h culture in media containing 30 mg Se4+∙L−1. The highest content of selenomethionine (238.8 μg∙gd.w.−1 was found after 48-h culture from the experimental medium that was supplemented with selenium at a concentration of 20 mg Se4+∙L−1. Biomass cell in the cultures supplemented with selenium ranged from 1.5 to 14.1 g∙L−1. The results of this study indicate that yeast cell biomass of C. utilis enriched mainly with the organic forms of selenium can be a valuable source of protein. It creates the possibility of obtaining selenium biocomplexes that can be used in the production of protein-selenium dietary supplements for animals and humans

Production of Newcastle Disease Virus by Vero Cells Grown on Cytodex 1 Microcarriers in a 2-Litre Stirred Tank Bioreactor

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Mohd Azmir Arifin

2010-01-01

Full Text Available The aim of this study is to prepare a model for the production of Newcastle disease virus (NDV lentogenic F strain using cell culture in bioreactor for live attenuated vaccine preparation. In this study, firstly we investigated the growth of Vero cells in several culture media. The maximum cell number was yielded by culture of Vero cells in Dulbecco's Modified Eagle Medium (DMEM which was 1.93×106 cells/ml. Secondly Vero cells were grown in two-litre stirred tank bioreactor by using several commercial microcarriers. We achieved the maximum cell concentration about 7.95×105 cells/ml when using Cytodex 1. Later we produced Newcastle Disease virus in stirred tank bioreactor based on the design developed using Taguchi L4 method. Results reveal that higher multiplicity of infection (MOI and size of cell inoculums can yield higher virus titer. Finally, virus samples were purified using high-speed centrifugation based on 3∗∗(3-1 Fractional Factorial Design. Statistical analysis showed that the maximum virus titer can be achieved at virus sample concentration of 58.45% (v/v, centrifugation speed of 13729 rpm, and centrifugation time of 4 hours. As a conclusion, high yield of virus titer could be achieved through optimization of cell culture in bioreactor and separation by high-speed centrifugation.

JSC technician checks STS-44 DSO 316 bioreactor and rotating wall vessel hdwr

Science.gov (United States)

1991-01-01

JSC technician Tacey Prewitt checks the progress on a bioreactor experiment in JSC's Life Sciences Laboratory Bldg 37 biotechnology laboratory. Similar hardware is scheduled for testing aboard Atlantis, Orbiter Vehicle (OV) 104, during STS-44. Detailed Supplementary Objective (DSO) 316 Bioreactor/Flow and Particle Trajectory in Microgravity will checkout the rotating wall vessel hardware and hopefully will confirm researchers' theories and calculations about how flow fields work in space. Plastic beads of various sizes rather than cell cultures are being flown in the vessel for the STS-44 test.

Control of an air pressure actuated disposable bioreactor for cultivating heart valves

NARCIS (Netherlands)

Beelen, M.J.; Neerincx, P.E.; Molengraft, van de M.J.G.

2011-01-01

A disposable injection molded bioreactor for growing tissue-engineered heart valves is controlled to mimic the physiological heart cycle. Tissue-engineered heart valves, cultured from human stem cells, are a possible alternative for replacing failing aortic heart valves, where nowadays biological

Bacterial study of the anaerobic bioreactor for distillery effluent

International Nuclear Information System (INIS)

Shah, F. A.; Pathan, M. I.

2006-01-01

This study relates with anaerobic bioreactors of Habib Sugar Mills, Nawabshah. Bacterial growth was studied through microscope along with its effect on the production of methane gas (Biogas) at all HRTs (Hydraulic Retention Times) between 15 and 28 days. The bacterium has the efficiency to convert 12% glucose within 24 hours to final product and cell mass. The acetogenic organisms also show their maximum growth on glucose in BGP-1 and BPG-2 at both the corks, where as Methanogenic organisms have shown their zero shown their zero growth on glucose. The efforts have been taken to determine the methanogenic, acetogenic and syntrophomonas sp. data of anaerobic bioreactors of BGP (Biogas Plant) I and II, when these samples were cultured on acetate, methanol, formate, butyrate, propionate and glucose. (author)

Biogas Production from Citrus Waste by Membrane Bioreactor

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Rachma Wikandari

2014-08-01

Full Text Available Rapid acidification and inhibition by d-limonene are major challenges of biogas production from citrus waste. As limonene is a hydrophobic chemical, this challenge was encountered using hydrophilic polyvinylidine difluoride (PVDF membranes in a biogas reactor. The more sensitive methane-producing archaea were encapsulated in the membranes, while freely suspended digesting bacteria were present in the culture as well. In this membrane bioreactor (MBR, the free digesting bacteria digested the citrus wastes and produced soluble compounds, which could pass through the membrane and converted to biogas by the encapsulated cell. As a control experiment, similar digestions were carried out in bioreactors containing the identical amount of just free cells. The experiments were carried out in thermophilic conditions at 55 °C, and hydraulic retention time of 30 days. The organic loading rate (OLR was started with 0.3 kg VS/m3/day and gradually increased to 3 kg VS/m3/day. The results show that at the highest OLR, MBR was successful to produce methane at 0.33 Nm3/kg VS, while the traditional free cell reactor reduced its methane production to 0.05 Nm3/kg VS. Approximately 73% of the theoretical methane yield was achieved using the membrane bioreactor.

Response coefficient analysis of a fed-batch bioreactor to dissolved oxygen perturbation in complementary cultures during PHB production

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Patnaik Pratap R

2008-03-01

Full Text Available Abstract Background Although the production of poly-β-hydroxybutyrate (PHB has many biological, energetic and environmental advantages over chemically synthesized polymers, synthetic polymers continue to be produced industrially since the productivities of fermentation processes fr PHB are not yet economically competitive. Improvement of a PHB fermentation requires good understanding and optimization under the realistic conditions of large bioreactors. Laboratory-scale studies have shown that co-cultures of Ralstonia eutropha and Lactobacillus delbrueckii generate better fermentation efficiencies than R. eutropha alone. In large bioreactors, incomplete dispersioin and perturbations in the dissolved oxygen (DO concentration, both of which affect the fermentation, have to be considered. This study analyzes the effect of DO fluctuations on bioreactor performance for both ideal and optimally dispersed broths. Results Response coefficient analysis was employed to obtain quantitative information on the effect of DO perturbations on different variables. Three values of the Peclet number (Pe cheracterized three levels of dispersion: Pe = 0.01 for nearly complete dispersion, Pe = 20 for optimum dispersion and Pe = 60 for insufficient dispersion. The response coefficients (RCs of the pairs of bacterial concentrations and the main substrates, glucose and ammonium chloride, showed contrasting variations with time. Lactate, a critical intermediate, and PHB had similar RC profiles but those of lactate were one to two orders of magnitude larger than other RCs. Significantly, the optimum Pe also resulted in the largest RCs, suggesting a balance between productivity and reactor stability. Conclusion Since R. eutropha requires oxygen for its growth whereas L. delbrueckii does not, fluctuations in the DO concentartion have a strong influence on the fermentation. Apart from this, the mechanism of PHB biosynthesis indicates that control of lactate is a critical

Hydrostatic pressure and shear stress affect endothelin-1 and nitric oxide release by endothelial cells in bioreactors.

Science.gov (United States)

Vozzi, Federico; Bianchi, Francesca; Ahluwalia, Arti; Domenici, Claudio

2014-01-01

Abundant experimental evidence demonstrates that endothelial cells are sensitive to flow; however, the effect of fluid pressure or pressure gradients that are used to drive viscous flow is not well understood. There are two principal physical forces exerted on the blood vessel wall by the passage of intra-luminal blood: pressure and shear. To analyze the effects of pressure and shear independently, these two stresses were applied to cultured cells in two different types of bioreactors: a pressure-controlled bioreactor and a laminar flow bioreactor, in which controlled levels of pressure or shear stress, respectively, can be generated. Using these bioreactor systems, endothelin-1 (ET-1) and nitric oxide (NO) release from human umbilical vein endothelial cells were measured under various shear stress and pressure conditions. Compared to the controls, a decrease of ET-1 production by the cells cultured in both bioreactors was observed, whereas NO synthesis was up-regulated in cells under shear stress, but was not modulated by hydrostatic pressure. These results show that the two hemodynamic forces acting on blood vessels affect endothelial cell function in different ways, and that both should be considered when planning in vitro experiments in the presence of flow. Understanding the individual and synergic effects of the two forces could provide important insights into physiological and pathological processes involved in vascular remodeling and adaptation. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Tylosin production by Streptomyces fradiae using raw cornmeal in airlift bioreactor.

Science.gov (United States)

Choi, Dubok; Choi, On You; Shin, Hyun-Jae; Chung, Dong-Ok; Shin, Dae-Yewn

2007-07-01

Using a 50-l airlift bioreactor, for the effective production of tylosin from Streptomyces fradiae TM-224 using raw cornmeal as the energy source, various environmental factors were studied in flask cultures. The maximum tylosin concentration was obtained at 32 degrees C and pH between 7.0 and 7.5. When seed was inoculated after 24 h of culture, the maximum tylosin concentration, 5.7 g/l, was obtained after 4 days of culture. Various concentrations of raw cornmeal were tested to investigate the optimum initial concentration for the tylosin production. An initial raw cornmeal concentration of 80 g/l gave the highest tylosin concentration, 5.8 g/l, after 5 days of culture. Of the various nitrogen sources, soybean meal and fish meal were found to be the most effective for the production of tylosin. In particular, with the optimal mixing ratio, 12 g/l of soybean meal to 14 g/l of fish meal, 7.2 g/l of tylosin was obtained after 5 days of culture. To compare raw cornmeal and glucose for the production oftylosin in the 50-1 airlift bioreactor for 10 days, fed-batch cultures were carried out under the optimum culture conditions. When raw corn meal was used as the energy source, the tylosin production increased with increasing culture time. The maximum tylosin concentration after 10 days of culture was 13.5 g/l, with a product yield from raw cornmeal of 0.123 g/g of consumed carbon source, which was about 7.2 times higher than that obtained when glucose was used as the carbon source.

Following an Optimal Batch Bioreactor Operations Model

DEFF Research Database (Denmark)

Ibarra-Junquera, V.; Jørgensen, Sten Bay; Virgen-Ortíz, J.J.

2012-01-01

The problem of following an optimal batch operation model for a bioreactor in the presence of uncertainties is studied. The optimal batch bioreactor operation model (OBBOM) refers to the bioreactor trajectory for nominal cultivation to be optimal. A multiple-variable dynamic optimization of fed...... as the master system which includes the optimal cultivation trajectory for the feed flow rate and the substrate concentration. The “real” bioreactor, the one with unknown dynamics and perturbations, is considered as the slave system. Finally, the controller is designed such that the real bioreactor...

Repairing the Osteochondral Defect in Goat with the Tissue-Engineered Osteochondral Graft Preconstructed in a Double-Chamber Stirring Bioreactor

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Yang Pei

2014-01-01

Full Text Available To investigate the reparative efficacy of tissue-engineered osteochondral (TEO graft for repairing the osteochondral defect in goat, we designed a double-chamber stirring bioreactor to construct the bone and cartilage composites simultaneously in one β-TCP scaffold and observed the reparative effect in vivo. The osteochondral defects were created in goats and all the animals were divided into 3 groups randomly. In groups A, the defect was treated with the TEO which was cultured with mechanical stimulation of stir; in group B, the defect was treated with TEO which was cultured without mechanical stimulation of stir; in groups C, the defect was treated without TEO. At 12 weeks and 24 weeks after operation, the reparative effects in different groups were assessed and compared. The results indicated that the reparative effect of the TEO cultured in the bioreactor was better than the control group, and mechanical stimulation of stir could further improve the reparative effect. We provided a feasible and effective method to construct the TEO for treatment of osteochondral defect using autologous BMSCs and the double-chamber bioreactor.

Dependence of synchronized bursting activity on medium stirring and the perfusion rate in a cultured network of neurons

Science.gov (United States)

Heo, Ryoun; Kim, Hyun; Lee, Kyoung J.

2016-05-01

A cultured network of neurons coupled with a multi-electrode-array (MEA) recording system has been a useful platform for investigating various issues in neuroscience and engineering. The neural activity supported by the system can be sensitive to environmental fluctuations, for example, in the medium's nutrient composition, ph, and temperature, and to mechanical disturbances, yet this issue has not been the subject. Especially, a normal practice in maintaining neuronal cell cultures involves an intermittent sequence of medium exchanges, typically at a time interval of a few days, and one such sudden medium exchange is unavoidably accompanied by many unintended disturbances. Here, based on a quantitative time-series analysis of synchronized bursting events, we explicitly demonstrate that such a medium exchange can, indeed, bring a huge change in the existing neural activity. Subsequently, we develop a medium perfusion-stirring system and an ideal protocol that can be used in conjunction with a MEA recording system, providing long-term stability. Specifically, we systematically evaluate the effects of medium stirring and perfusion rates. Unexpectedly, even some vigorous mechanical agitations do not have any impacts on neural activity. On the other hand, too much replenishment ( e.g., 1.8 ml/day for a 1.8-ml dish) of neurobasal medium results in an excitotoxicity.

Characterization and Application of a Disposable Rotating Bed Bioreactor for Mesenchymal Stem Cell Expansion.

Science.gov (United States)

Neumann, Anne; Lavrentieva, Antonina; Heilkenbrinker, Alexandra; Loenne, Maren; Kasper, Cornelia

2014-11-27

Recruitment of mesenchymal stromal cells (MSC) into the field of tissue engineering is a promising development since these cells can be expanded vivo to clinically relevant numbers and, after expansion, retain their ability to differentiate into various cell lineages. Safety requirements and the necessity to obtain high cell numbers without frequent subcultivation of cells raised the question of the possibility of expanding MSC in one-way (single-use) disposable bioreactors. In this study, umbilical cord-derived MSC (UC-MSC) were expanded in a disposable Z 2000 H bioreactor under dynamic conditions. Z was characterized regarding residence time and mixing in order to evaluate the optimal bioreactor settings, enabling optimal mass transfer in the absence of shear stress, allowing an reproducible expansion of MSC, while maintaining their stemness properties. Culture of the UC-MSC in disposable Z 2000 H bioreactor resulted in a reproducible 8-fold increase of cell numbers after 5 days. Cells were shown to maintain specific MSC surface marker expression as well as trilineage differentiation potential and lack stress-induced premature senescence.

Characterization and Application of a Disposable Rotating Bed Bioreactor for Mesenchymal Stem Cell Expansion

Directory of Open Access Journals (Sweden)

Anne Neumann

2014-11-01

Full Text Available Recruitment of mesenchymal stromal cells (MSC into the field of tissue engineering is a promising development since these cells can be expanded vivo to clinically relevant numbers and, after expansion, retain their ability to differentiate into various cell lineages. Safety requirements and the necessity to obtain high cell numbers without frequent subcultivation of cells raised the question of the possibility of expanding MSC in one-way (single-use disposable bioreactors. In this study, umbilical cord-derived MSC (UC-MSC were expanded in a disposable Z 2000 H bioreactor under dynamic conditions. Z was characterized regarding residence time and mixing in order to evaluate the optimal bioreactor settings, enabling optimal mass transfer in the absence of shear stress, allowing an reproducible expansion of MSC, while maintaining their stemness properties. Culture of the UC-MSC in disposable Z 2000 H bioreactor resulted in a reproducible 8-fold increase of cell numbers after 5 days. Cells were shown to maintain specific MSC surface marker expression as well as trilineage differentiation potential and lack stress-induced premature senescence.

Engineering Parameters in Bioreactor's Design: A Critical Aspect in Tissue Engineering

Science.gov (United States)

Amoabediny, Ghassem; Pouran, Behdad; Tabesh, Hadi; Shokrgozar, Mohammad Ali; Haghighipour, Nooshin; Khatibi, Nahid; Mottaghy, Khosrow; Zandieh-Doulabi, Behrouz

2013-01-01

Bioreactors are important inevitable part of any tissue engineering (TE) strategy as they aid the construction of three-dimensional functional tissues. Since the ultimate aim of a bioreactor is to create a biological product, the engineering parameters, for example, internal and external mass transfer, fluid velocity, shear stress, electrical current distribution, and so forth, are worth to be thoroughly investigated. The effects of such engineering parameters on biological cultures have been addressed in only a few preceding studies. Furthermore, it would be highly inefficient to determine the optimal engineering parameters by trial and error method. A solution is provided by emerging modeling and computational tools and by analyzing oxygen, carbon dioxide, and nutrient and metabolism waste material transports, which can simulate and predict the experimental results. Discovering the optimal engineering parameters is crucial not only to reduce the cost and time of experiments, but also to enhance efficacy and functionality of the tissue construct. This review intends to provide an inclusive package of the engineering parameters together with their calculation procedure in addition to the modeling techniques in TE bioreactors. PMID:24000327

Engineering parameters in bioreactor's design: a critical aspect in tissue engineering.

Science.gov (United States)

Salehi-Nik, Nasim; Amoabediny, Ghassem; Pouran, Behdad; Tabesh, Hadi; Shokrgozar, Mohammad Ali; Haghighipour, Nooshin; Khatibi, Nahid; Anisi, Fatemeh; Mottaghy, Khosrow; Zandieh-Doulabi, Behrouz

2013-01-01

Bioreactors are important inevitable part of any tissue engineering (TE) strategy as they aid the construction of three-dimensional functional tissues. Since the ultimate aim of a bioreactor is to create a biological product, the engineering parameters, for example, internal and external mass transfer, fluid velocity, shear stress, electrical current distribution, and so forth, are worth to be thoroughly investigated. The effects of such engineering parameters on biological cultures have been addressed in only a few preceding studies. Furthermore, it would be highly inefficient to determine the optimal engineering parameters by trial and error method. A solution is provided by emerging modeling and computational tools and by analyzing oxygen, carbon dioxide, and nutrient and metabolism waste material transports, which can simulate and predict the experimental results. Discovering the optimal engineering parameters is crucial not only to reduce the cost and time of experiments, but also to enhance efficacy and functionality of the tissue construct. This review intends to provide an inclusive package of the engineering parameters together with their calculation procedure in addition to the modeling techniques in TE bioreactors.

Modular 3D printed lab-on-a-chip bio-reactor for the biochemical energy cascade of microorganisms

Science.gov (United States)

Podwin, Agnieszka; Dziuban, Jan A.

2017-10-01

The paper presents the sandwiched polymer 3D printed lab-on-a-chip bio-reactor for the biochemical energy cascade of microorganisms. Euglenas and yeast were separately and simultaneously cultured for 10 d in the chip. As a result of the experiments, euglenas, light-initialized and nourished by CO2—a product of ethanol fermentation handled by yeast—generated oxygen, based on the photosynthesis process. The presence of oxygen in the bio-reactor was confirmed by the colorimetric method—a bicarbonate (pH) indicator. Preliminary studies towards the obtainment of an effective source of oxygen are promising and further research should be done to enable the utility of the bio-reactor in, for instance, microbial fuel cells.

Modular 3D printed lab-on-a-chip bio-reactor for the biochemical energy cascade of microorganisms

International Nuclear Information System (INIS)

Podwin, Agnieszka; Dziuban, Jan A

2017-01-01

The paper presents the sandwiched polymer 3D printed lab-on-a-chip bio-reactor for the biochemical energy cascade of microorganisms. Euglenas and yeast were separately and simultaneously cultured for 10 d in the chip. As a result of the experiments, euglenas, light-initialized and nourished by CO 2 —a product of ethanol fermentation handled by yeast—generated oxygen, based on the photosynthesis process. The presence of oxygen in the bio-reactor was confirmed by the colorimetric method—a bicarbonate (pH) indicator. Preliminary studies towards the obtainment of an effective source of oxygen are promising and further research should be done to enable the utility of the bio-reactor in, for instance, microbial fuel cells. (paper)

Production and release of infectious hepatitis C virus from human liver cell cultures in the three-dimensional radial-flow bioreactor

International Nuclear Information System (INIS)

Aizaki, Hideki; Nagamori, Seishi; Matsuda, Mami; Kawakami, Hayato; Hashimoto, Osamu; Ishiko, Hiroaki; Kawada, Masaaki; Matsuura, Tomokazu; Hasumura, Satoshi; Matsuura, Yoshiharu; Suzuki, Tetsuro; Miyamura, Tatsuo

2003-01-01

Lack of efficient culture systems for hepatitis C virus (HCV) has been a major obstacle in HCV research. Human liver cells grown in a three-dimensional radial-flow bioreactor were successfully infected following inoculation with plasma from an HCV carrier. Subsequent detection of increased HCV RNA suggested viral replication. Furthermore, transfection of HCV RNA transcribed from full-length cDNA also resulted in the production and release of HCV virions into supernatant. Infectivity was shown by successful secondary passage to a new culture. Introduction of mutations in RNA helicase and polymerase regions of HCV cDNA abolished virus replication, indicating that reverse genetics of this system is possible. The ability to replicate and detect the extracellular release of HCV might provide clues with regard to the persistent nature of HCV infection. It will also accelerate research into the pathogenicity of HCV, as well as the development of prophylactic agents and new therapy

Producing Newborn Synchronous Mammalian Cells

Science.gov (United States)

Gonda, Steve R.; Helmstetter, Charles E.; Thornton, Maureen

2008-01-01

A method and bioreactor for the continuous production of synchronous (same age) population of mammalian cells have been invented. The invention involves the attachment and growth of cells on an adhesive-coated porous membrane immersed in a perfused liquid culture medium in a microgravity analog bioreactor. When cells attach to the surface divide, newborn cells are released into the flowing culture medium. The released cells, consisting of a uniform population of synchronous cells are then collected from the effluent culture medium. This invention could be of interest to researchers investigating the effects of the geneotoxic effects of the space environment (microgravity, radiation, chemicals, gases) and to pharmaceutical and biotechnology companies involved in research on aging and cancer, and in new drug development and testing.

Establishing Early Functional Perfusion and Structure in Tissue Engineered Cardiac Constructs.

Science.gov (United States)

Wang, Bo; Patnaik, Sourav S; Brazile, Bryn; Butler, J Ryan; Claude, Andrew; Zhang, Ge; Guan, Jianjun; Hong, Yi; Liao, Jun

2015-01-01

Myocardial infarction (MI) causes massive heart muscle death and remains a leading cause of death in the world. Cardiac tissue engineering aims to replace the infarcted tissues with functional engineered heart muscles or revitalize the infarcted heart by delivering cells, bioactive factors, and/or biomaterials. One major challenge of cardiac tissue engineering and regeneration is the establishment of functional perfusion and structure to achieve timely angiogenesis and effective vascularization, which are essential to the survival of thick implants and the integration of repaired tissue with host heart. In this paper, we review four major approaches to promoting angiogenesis and vascularization in cardiac tissue engineering and regeneration: delivery of pro-angiogenic factors/molecules, direct cell implantation/cell sheet grafting, fabrication of prevascularized cardiac constructs, and the use of bioreactors to promote angiogenesis and vascularization. We further provide a detailed review and discussion on the early perfusion design in nature-derived biomaterials, synthetic biodegradable polymers, tissue-derived acellular scaffolds/whole hearts, and hydrogel derived from extracellular matrix. A better understanding of the current approaches and their advantages, limitations, and hurdles could be useful for developing better materials for future clinical applications.

Bioreactor engineering of stem cell environments.

Science.gov (United States)

Tandon, Nina; Marolt, Darja; Cimetta, Elisa; Vunjak-Novakovic, Gordana

2013-11-15

Stem cells hold promise to revolutionize modern medicine by the development of new therapies, disease models and drug screening systems. Standard cell culture systems have limited biological relevance because they do not recapitulate the complex 3-dimensional interactions and biophysical cues that characterize the in vivo environment. In this review, we discuss the current advances in engineering stem cell environments using novel biomaterials and bioreactor technologies. We also reflect on the challenges the field is currently facing with regard to the translation of stem cell based therapies into the clinic. Copyright © 2013 Elsevier Inc. All rights reserved.

The Effect of Primary Cancer Cell Culture Models on the Results of Drug Chemosensitivity Assays: The Application of Perfusion Microbioreactor System as Cell Culture Vessel

Science.gov (United States)

Chen, Yi-Dao; Huang, Shiang-Fu; Wang, Hung-Ming

2015-01-01

To precisely and faithfully perform cell-based drug chemosensitivity assays, a well-defined and biologically relevant culture condition is required. For the former, a perfusion microbioreactor system capable of providing a stable culture condition was adopted. For the latter, however, little is known about the impact of culture models on the physiology and chemosensitivity assay results of primary oral cavity cancer cells. To address the issues, experiments were performed. Results showed that minor environmental pH change could significantly affect the metabolic activity of cells, demonstrating the importance of stable culture condition for such assays. Moreover, the culture models could also significantly influence the metabolic activity and proliferation of cells. Furthermore, the choice of culture models might lead to different outcomes of chemosensitivity assays. Compared with the similar test based on tumor-level assays, the spheroid model could overestimate the drug resistance of cells to cisplatin, whereas the 2D and 3D culture models might overestimate the chemosensitivity of cells to such anticancer drug. In this study, the 3D culture models with same cell density as that in tumor samples showed comparable chemosensitivity assay results as the tumor-level assays. Overall, this study has provided some fundamental information for establishing a precise and faithful drug chemosensitivity assay. PMID:25654105

Continuous production of lactic acid from molasses by perfusion culture of Lactococcus lactis using a stirred ceramic membrane reactor.

Science.gov (United States)

Ohashi, R; Yamamoto, T; Suzuki, T

1999-01-01

A perfusion culture system was used for continuous production of lactic acid by retaining cells at a high density of Lactococcus lactis in a stirred ceramic membrane reactor (SCMR). After the cell concentration increased to 248 g/l, half of the culture broth volume was replaced with the fermentation medium. Subsequently, a substrate solution containing glucose (run 1) or molasses (run 2) was continuously supplied to the cells retained in the SCMR. Simultaneously, the culture supernatant was extracted using a ceramic filter with a pore size of 0.2 mum. The dilution rate was initially set at 0.4 h(-1) and gradually decreased to 0.2 h(-1) due to reduction in the permeability of the filter. The concentration of glucose in the substrate solution was adjusted to 60 g/l for the transition and the first period until 240 h, 90 g/l for the second period from 240 h to 440 h, and 70 g/l for the third period from 440 h to 643 h. The average concentration of lactic acid in the filtrate reached 46 g/l in the first period, 43 g/l in the second period, and 33 g/l for the third period. The productivity obtained for the first period reached 15.8 g.l(-1).h(-1), twice as much as that achieved in repeated batch fermentations. Based on the results obtained in run 1, the substrate solution containing 120 g/l of molasses was continuously supplied for 240 h in run 2. The concentration and productivity of lactic acid reached 40 g/l and 10.6 g.l(-1).h(-1), respectively, by continuously replenishing the culture medium at a dilution rate of 0.26 h(-1). These results demonstrated that the filtration capacity of the SCMR was sufficient for a continuous and rapid replenishment of molasses solution from the dense cell culture and, therefore, the perfusion culture system is considered to provide a low-cost process for continuous production of lactic acid from cheap resources.

Biological reduction of nitrates in wastewaters from nuclear processing using a fluidized-bed bioreactor

International Nuclear Information System (INIS)

Pitt, W.W.; Hancher, C.W.; Patton, B.D.

1981-01-01

There are a number of nitrate-containing wastewater sources, as concentrated as 30 wt.% NO 3 - and as large as 2000 m 3 /day, in the nuclear fuel cycle. The biological reduction of nitrate in wastewater to gaseous nitrogen, accompanied by the oxidation of a nutrient carbon source to gaseous carbon dioxide, is an ecologically sound and cost-effective method of treating wastewaters containing nitrates. These nitrate-containing wastewater sources can be successfully biologically denitrified to meet discharge standards in the range of 10 to 20 gN(NO 3 - )/m 3 by the use of a fluidized-bed bioreactor. The denitrification bacteria are a mixed culture derived from garden soil; the major strain is Pseudomonas. In the fluidized-bed bioreactor the bacteria are allowed to attach to 0.25- to 0.50-mm-diam coal fluidization particles, which are then fluidized by the upward flow of influent wastewater. Maintaining the bacteria-to-coal weight ratio at approximately 1:10 results in a bioreactor bacteria loading of greater than 20,000 g/m 3 . This paper describes the results of a biodenitrification R and D program based on the use of fluidized bioreactors capable of operating at nitrate levels up to 7000 g/m 3 and achieving denitrification rates as high as 80 g N(NO 3 - ) per day per liter of empty bioreactor volume. 4 figures, 7 tables

Dynamics of yeast immobilized-cell fluidized-bed bioreactors systems in ethanol fermentation from lactose-hydrolyzed whey and whey permeate.

Science.gov (United States)

Gabardo, Sabrina; Pereira, Gabriela Feix; Klein, Manuela P; Rech, Rosane; Hertz, Plinho F; Ayub, Marco Antônio Záchia

2016-01-01

We studied the dynamics of ethanol production on lactose-hydrolyzed whey (LHW) and lactose-hydrolyzed whey permeate (LHWP) in batch fluidized-bed bioreactors using single and co-cultures of immobilized cells of industrial strains of Saccharomyces cerevisiae and non-industrial strains of Kluyveromyces marxianus. Although the co-culture of S. cerevisiae CAT-1 and K. marxianus CCT 4086 produced two- to fourfold the ethanol productivity of single cultures of S. cerevisiae, the single cultures of the K. marxianus CCT 4086 produced the best results in both media (Y EtOH/S = 0.47-0.49 g g(-1) and Q P = 1.39-1.68 g L(-1) h(-1), in LHW and LHWP, respectively). Ethanol production on concentrated LHWP (180 g L(-1)) reached 79.1 g L(-1), with yields of 0.46 g g(-1) for K. marxianus CCT 4086 cultures. Repeated batches of fluidized-bed bioreactor on concentrated LHWP led to increased ethanol productivity, reaching 2.8 g L(-1) h(-1).

Resveratrol production in bioreactor: Assessment of cell physiological states and plasmid segregational stability

Directory of Open Access Journals (Sweden)

Margarida S. Afonso

2015-03-01

Full Text Available Resveratrol is a plant secondary metabolite commonly found in peanuts and grapevines with significant health benefits. Recombinant organisms can produce large amounts of resveratrol and, in this work, Escherichia coli BW27784 was used to produce resveratrol in bioreactors while monitoring cell physiology and plasmid stability through flow cytometry and real-time qPCR, respectively. Initially, the influence of culture conditions and precursor addition was evaluated in screening assays and the data gathered was used to perform the bioreactor assays, allowing the production of 160 μg/mL of resveratrol. Cellular physiology and plasmid instability affected the final resveratrol production, with lower viability and plasmid copy numbers associated with lower yields. In sum, this study describes new tools to monitor the bioprocess, evaluating the effect of culture conditions, and its correlation with cell physiology and plasmid segregational stability, in order to define a viable and scalable bioprocess to fulfill the need for larger quantities of resveratrol.

Accelerated and Improved Differentiation of Retinal Organoids from Pluripotent Stem Cells in Rotating-Wall Vessel Bioreactors

Directory of Open Access Journals (Sweden)

Tyler DiStefano

2018-01-01

Full Text Available Pluripotent stem cells can be differentiated into 3D retinal organoids, with major cell types self-patterning into a polarized, laminated architecture. In static cultures, organoid development may be hindered by limitations in diffusion of oxygen and nutrients. Herein, we report a bioprocess using rotating-wall vessel (RWV bioreactors to culture retinal organoids derived from mouse pluripotent stem cells. Organoids in RWV demonstrate enhanced proliferation, with well-defined morphology and improved differentiation of neurons including ganglion cells and S-cone photoreceptors. Furthermore, RWV organoids at day 25 (D25 reveal similar maturation and transcriptome profile as those at D32 in static culture, closely recapitulating spatiotemporal development of postnatal day 6 mouse retina in vivo. Interestingly, however, retinal organoids do not differentiate further under any in vitro condition tested here, suggesting additional requirements for functional maturation. Our studies demonstrate that bioreactors can accelerate and improve organoid growth and differentiation for modeling retinal disease and evaluation of therapies.

On-line removal of volatile fatty acids from CELSS anaerobic bioreactor via nanofiltration

Science.gov (United States)

Colon, Guillermo

1995-01-01

The CELSS (controlled ecological life support system) resource recovery system, which is a waste processing system, uses aerobic and anaerobic bioreactors to recover plants nutrients and secondary foods from the inedible biomass. The anaerobic degradation of the inedible biomass by means of culture of rumen bacteria,generates organic compounds such as volatile fatty acids (acetic, propionic, butyric, VFA) and ammonia. The presence of VFA in the bioreactor medium at fairly low concentrations decreases the microbial population's metabolic reactions due to end-product inhibition. Technologies to remove VFA continuously from the bioreactor are of high interest. Several candidate technologies were analyzed, such as organic solvent liquid-liquid extraction, adsorption and/or ion exchange, dialysis, electrodialysis, and pressure driven membrane separation processes. The proposed technique for the on-line removal of VFA from the anaerobic bioreactor was a nanofiltration membrane recycle bioreactor. In order to establish the nanofiltration process performance variables before coupling it to the bioreactor, a series of experiments were carried out using a 10,000 MWCO tubular ceramic membrane module. The variables studied were the bioreactor slurry permeation characteristics, such as, the permeate flux, VFA and the nutrient removal rates as a function of applied transmembrane pressure, fluid recirculation velocity, suspended matter concentration, and process operating time. Results indicate that the permeate flux, VFA and nutrients removal rates are directly proportional to the fluid recirculation velocity in the range between 0.6 to 1.0 m/s, applied pressure when these are low than 1.5 bar, and inversely proportional to the total suspended solids concentration in the range between 23,466 to 34,880. At applied pressure higher than 1.5 bar the flux is not more linearly dependent due to concentration polarization and fouling effects over the membrange surface. It was also found

On-line removal of volatile fatty acids from CELSS anaerobic bioreactor via nanofiltration.

Science.gov (United States)

Colon, G; Sager, J C

2001-01-01

The CELSS resource recovery system, which is a waste-processing system, uses aerobic and anaerobic bioreactors to recover plants nutrients and secondary foods from the inedible biomass. The anaerobic degradation of the inedible biomass, by means of culture of rumen bacteria, generates organic compounds such as volatile fatty acids (VFA) (acetic, propionic, butyric) and ammonia. The presence of VFA in the bioreactor medium at fairly low concentrations decreases the microbial population's metabolic reactions due to end-product inhibition. Technologies to remove VFA continuously from the bioreactor are of high interest. Several candidate technologies were analyzed, such as organic solvent liquid-liquid extraction, adsorption and/or ion exchange, dialysis, electrodialysis, and pressure-driven membrane separation processes. The proposed technique for the on-line removal of VFA from the anaerobic bioreactor was a nanofiltration membrane recycle bioreactor. In order to establish the nanofiltration process performance variables before coupling it to the bioreactor, a series of experiments was carried out using a 10,000 molecular weight cutoff (MWCO) tubular ceramic membrane module. The variables studied were the bioreactor slurry permeation characteristics, such as: the permeate flux, VFA and nutrient removal rates as a function of applied transmembrane pressure, fluid recirculation velocity, suspended matter concentration, and process operating time. Results indicated that the permeate flux, VFA, and nutrients removal rates are directly proportional to the fluid recirculation velocity in the range between 0.6 and 1.0 m/s, applied pressure when these are lower than 1.5 bar, and inversely proportional to the total suspended solids concentration in the range between 23,466 and 34,880 mg/L. At applied pressure higher than 1.5 bar the flux is not more linearly dependent due to concentration polarization and fouling effects over the membrane surface. It was also found that the

Bioreactor design and optimization – a future perspective

DEFF Research Database (Denmark)

Gernaey, Krist

2011-01-01

Bioreactor design and optimisation are essential in translating the experience gained from lab or pilot scale experiments to efficient production processes in industrial scale bioreactors. This article gives a future perspective on bioreactor design and optimisation, where it is foreseen...

Scaled-up manufacturing of recombinant antibodies produced by plant cells in a 200-L orbitally-shaken disposable bioreactor.

Science.gov (United States)

Raven, Nicole; Rasche, Stefan; Kuehn, Christoph; Anderlei, Tibor; Klöckner, Wolf; Schuster, Flora; Henquet, Maurice; Bosch, Dirk; Büchs, Jochen; Fischer, Rainer; Schillberg, Stefan

2015-02-01

Tobacco BY-2 cells have emerged as a promising platform for the manufacture of biopharmaceutical proteins, offering efficient protein secretion, favourable growth characteristics and cultivation in containment under a controlled environment. The cultivation of BY-2 cells in disposable bioreactors is a useful alternative to conventional stainless steel stirred-tank reactors, and orbitally-shaken bioreactors could provide further advantages such as simple bag geometry, scalability and predictable process settings. We carried out a scale-up study, using a 200-L orbitally-shaken bioreactor holding disposable bags, and BY-2 cells producing the human monoclonal antibody M12. We found that cell growth and recombinant protein accumulation were comparable to standard shake flask cultivation, despite a 200-fold difference in cultivation volume. Final cell fresh weights of 300-387 g/L and M12 yields of ∼20 mg/L were achieved with both cultivation methods. Furthermore, we established an efficient downstream process for the recovery of M12 from the culture broth. The viscous spent medium prevented clarification using filtration devices, but we used expanded bed adsorption (EBA) chromatography with SP Sepharose as an alternative for the efficient capture of the M12 antibody. EBA was introduced as an initial purification step prior to protein A affinity chromatography, resulting in an overall M12 recovery of 75-85% and a purity of >95%. Our results demonstrate the suitability of orbitally-shaken bioreactors for the scaled-up cultivation of plant cell suspension cultures and provide a strategy for the efficient purification of antibodies from the BY-2 culture medium. © 2014 Wiley Periodicals, Inc.

Process technology of luwak coffee through bioreactor utilization

Science.gov (United States)

Hadipernata, M.; Nugraha, S.

2018-01-01

Indonesia has an advantage in producing exotic coffee that is Luwak coffee. Luwak coffee is produced from the fermentation process in digestion of civet. Luwak coffee production is still limited due to the difficulty level in the use of civet animals as the only medium of Luwak coffee making. The research was conducted by developing technology of luwak coffee production through bioreactor utilization and addition the bacteria isolate from gastric of civet. The process conditions in the bioreactor which include temperature, pH, and bacteria isolate of civet are adjusted to the process that occurs in civet digestion, including peristaltic movement on the stomach and small intestine of the civet will be replaced by the use of propellers that rotate on the bioreactor. The result of research showed that proximat analysis data of artificial/bioreactor luwak coffee did not significant different with original luwak coffee. However, the original luwak coffee has higher content of caffeine compared to bioreactor luwak coffee. Based on the cuping test the bioreactor luwak coffee has a value of 84.375, while the original luwak coffee is 84.875. As the result, bioreactor luwak coffee has excellent taste that similiar with original luwak coffee taste.

Perfusion-induced changes in cardiac contractility depend on capillary perfusion.

Science.gov (United States)

Dijkman, M A; Heslinga, J W; Sipkema, P; Westerhof, N

1998-02-01

The perfusion-induced increase in cardiac contractility (Gregg phenomenon) is especially found in heart preparations that lack adequate coronary autoregulation and thus protection of changes in capillary pressure. We determined in the isolated perfused papillary muscle of the rat whether cardiac muscle contractility is related to capillary perfusion. Oxygen availability of this muscle is independent of internal perfusion, and perfusion may be varied or even stopped without loss of function. Muscles contracted isometrically at 27 degrees C (n = 7). During the control state stepwise increases in perfusion pressure resulted in all muscles in a significant increase in active tension. Muscle diameter always increased with increased perfusion pressure, but muscle segment length was unaffected. Capillary perfusion was then obstructed by plastic microspheres (15 microns). Flow, at a perfusion pressure of 66.6 +/- 26.2 cmH2O, reduced from 17.6 +/- 5.4 microliters/min in the control state to 3.2 +/- 1.3 microliters/min after microspheres. Active tension developed by the muscle in the unperfused condition before microspheres and after microspheres did not differ significantly (-12.8 +/- 29.4% change). After microspheres similar perfusion pressure steps as in control never resulted in an increase in active tension. Even at the two highest perfusion pressures (89.1 +/- 28.4 and 106.5 +/- 31.7 cmH2O) that were applied a significant decrease in active tension was found. We conclude that the Gregg phenomenon is related to capillary perfusion.

Micro propagation of Stevia rebaudiana Bertoni through temporary immersion bioreactor system

International Nuclear Information System (INIS)

Norazlina Noordin; Rusli Ibrahim; Nur Hidayah Sajahan; Siti Maryam Mohd Nahar; Siti Hajar Mohd Nahar

2012-01-01

Stevia rebaudiana Bertoni is a perennial herb that belongs to the family of Asteraceae. It is a natural sweetener plant known as sweet leaf, which is estimated to be 300 times sweeter than cane sugar. In this study, micro propagation of this natural herb via temporary immersion bioreactor system was successfully conducted. Shoot tips and nodal segment were used as explants to induce multiply shoots. It was found that shoot tips on MS medium supplemented with 1 mg/l Kinetin showed the highest shoot multiplication after 3 weeks of culture. Shoot elongation and rooting was successfully optimized in MS basal medium 2 weeks later. Mass propagation of stevia shoots were carried out in temporary immersion bioreactor and this system showed promising potential as an alternative approach for rapid and continuous production of in vitro stevia plantlets. (author)

Hepatic perfusion during hepatic artery infusion chemotherapy: Evaluation with perfusion CT and perfusion scintigraphy

International Nuclear Information System (INIS)

Miller, D.L.; Carrasquillo, J.A.; Lutz, R.J.; Chang, A.E.

1989-01-01

The standard method for the evaluation of hepatic perfusion during hepatic artery infusion (HAI) chemotherapy is planar hepatic artery perfusion scintigraphy (HAPS). Planar HAPS was performed with 2 mCi of [99mTc] macroaggregated albumin infused at 1 ml/min and compared with single photon emission CT (SPECT) HAPS and with a new study, CT performed during the slow injection of contrast material through the HAI catheter (HAI-CT). Thirteen patients underwent 16 HAI-CT studies, 14 planar HAPS studies, and 9 SPECT HAPS studies. In 13 of 14 studies (93%) HAI-CT and planar HAPS were in complete agreement as to the perfusion pattern of intrahepatic metastases and normal liver. In nine studies where all modalities were performed, the findings identified by HAI-CT and planar HAPS agreed in all cases, whereas the results of two SPECT scans disagreed with the other studies. With respect to perfusion of individual metastases, 14 of 14 HAI-CT studies, 12 of 13 planar HAPS studies, and 9 of 9 SPECT HAPS studies correctly demonstrated the perfusion status of individual lesions as indicated by the pattern of changes in tumor size determined on CT obtained before and after the perfusion studies. Hepatic artery infusion CT was superior for delineation of individual metastases, particularly small lesions, and for the evaluation of nonperfused portions of the liver. Planar HAPS detected extrahepatic perfusion in four patients, and this was not detected by HAI-CT. We conclude that HAI-CT and scintigraphy are complementary techniques. Hepatic artery infusion CT has advantages for the evaluation of intrahepatic perfusion, and planar HAPS is superior to HAI-CT for the detection of extrahepatic perfusion

Impact of scaffold micro and macro architecture on Schwann cell proliferation under dynamic conditions in a rotating wall vessel bioreactor

International Nuclear Information System (INIS)

Valmikinathan, Chandra M.; Hoffman, John; Yu, Xiaojun

2011-01-01

Over the last decade tissue engineering has emerged as a powerful alternative to regenerate lost tissues owing to trauma or tumor. Evidence shows that Schwann cell containing scaffolds have improved performance in vivo as compared to scaffolds that depend on cellularization post implantation. However, owing to limited supply of cells from the patients themselves, several approaches have been taken to enhance cell proliferation rates to produce complete and uniform cellularization of scaffolds. The most common approach is the application of a bioreactor to enhance cell proliferation rate and therefore reduce the time needed to obtain sufficiently significant number of glial cells, prior to implantation. In this study, we show the application of a rotating wall bioreactor system for studying Schwann cell proliferation on nanofibrous spiral shaped scaffolds, prepared by solvent casting and salt leaching techniques. The scaffolds were fabricated from polycaprolactone (PCL), which has ideal mechanical properties and upon degradation does not produce acidic byproducts. The spiral scaffolds were coated with aligned or random nanofibers, produced by electrospinning, to provide a substrate that mimics the native extracellular matrix and the essential contact guidance cues. At the 4 day time point, an enhanced rate of cell proliferation was observed on the open structured nanofibrous spiral scaffolds in a rotating wall bioreactor, as compared to static culture conditions. However, the cell proliferation rate on the other contemporary scaffolds architectures such as the tubular and cylindrical scaffolds show reduced cell proliferation in the bioreactor as compared to static conditions, at the same time point. Moreover, the rotating wall bioreactor does not alter the orientation or the phenotype of the Schwann cells on the aligned nanofiber containing scaffolds, wherein, the cells remain aligned along the length of the scaffolds. Therefore, these open structured spiral

Capacity Planning for Batch and Perfusion Bioprocesses Across Multiple Biopharmaceutical Facilities

OpenAIRE

Siganporia, Cyrus C; Ghosh, Soumitra; Daszkowski, Thomas; Papageorgiou, Lazaros G; Farid, Suzanne S

2014-01-01

Production planning for biopharmaceutical portfolios becomes more complex when products switch between fed-batch and continuous perfusion culture processes. This article describes the development of a discrete-time mixed integer linear programming (MILP) model to optimize capacity plans for multiple biopharmaceutical products, with either batch or perfusion bioprocesses, across multiple facilities to meet quarterly demands. The model comprised specific features to account for products with fe...

Membrane bioreactors for waste gas treatment.

NARCIS (Netherlands)

Reij, M.W.; Keurentjes, J.T.F.; Hartmans, S.

1998-01-01

This review describes the recent development of membrane reactors for biological treatment of waste gases. In this type of bioreactor gaseous pollutants are transferred through a membrane to the liquid phase, where micro-organisms degrade the pollutants. The membrane bioreactor combines the

Membrane bioreactors for waste gas treatment

NARCIS (Netherlands)

Reij, M.W.; Keurentjes, J.T.F.; Hartmans, S.

1998-01-01

This review describes the recent development of membrane reactors for biological treatment of waste gases. In this type of bioreactor gaseous pollutants are transferred through a membrane to the liquid phase, where micro-organisms degrade the pollutants. The membrane bioreactor combines the

Fabrication, characterization, and in vitro evaluation of poly(lactic acid glycolic acid)/nano-hydroxyapatite composite microsphere-based scaffolds for bone tissue engineering in rotating bioreactors.

Science.gov (United States)

Lv, Qing; Nair, Lakshmi; Laurencin, Cato T

2009-12-01

Dynamic flow culture bioreactor systems have been shown to enhance in vitro bone tissue formation by facilitating mass transfer and providing mechanical stimulation. Our laboratory has developed a biodegradable poly (lactic acid glycolic acid) (PLAGA) mixed scaffold consisting of lighter-than-water (LTW) and heavier-than-water (HTW) microspheres as potential matrices for engineering tissue using a high aspect ratio vessel (HARV) rotating bioreactor system. We have demonstrated enhanced osteoblast differentiation and mineralization on PLAGA scaffolds in the HARV rotating bioreactor system when compared with static culture. The objective of the present study is to improve the mechanical properties and bioactivity of polymeric scaffolds by designing LTW polymer/ceramic composite scaffolds suitable for dynamic culture using a HARV bioreactor. We employed a microsphere sintering method to fabricate three-dimensional PLAGA/nano-hydroxyapatite (n-HA) mixed scaffolds composed of LTW and HTW composite microspheres. The mechanical properties, pore size and porosity of the composite scaffolds were controlled by varying parameters, such as sintering temperature, sintering time, and PLAGA/n-HA ratio. The PLAGA/n-HA (4:1) scaffold sintered at 90 degrees C for 3 h demonstrated the highest mechanical properties and an appropriate pore structure for bone tissue engineering applications. Furthermore, evaluation human mesenchymal stem cells (HMSCs) response to PLAGA/n-HA scaffolds was performed. HMSCs on PLAGA/n-HA scaffolds demonstrated enhanced proliferation, differentiation, and mineralization when compared with those on PLAGA scaffolds. Therefore, PLAGA/n-HA mixed scaffolds are promising candidates for HARV bioreactor-based bone tissue engineering applications. Copyright 2008 Wiley Periodicals, Inc.

Role of Bioreactors in Microbial Biomass and Energy Conversion

Energy Technology Data Exchange (ETDEWEB)

Zhang, Liang [Chongqing University, Chongqing, China; Zhang, Biao [Chongqing University, Chongqing, China; Zhu, Xun [Chongqing University, Chongqing, China; Chang, Haixing [Chongqing University of Technology; Ou, Shawn [ORNL; Wang, HONG [Chongqing University, Chongqing, China

2018-04-01

Bioenergy is the world’s largest contributor to the renewable and sustainable energy sector, and it plays a significant role in various energy industries. A large amount of research has contributed to the rapidly evolving field of bioenergy and one of the most important topics is the use of the bioreactor. Bioreactors play a critical role in the successful development of technologies for microbial biomass cultivation and energy conversion. In this chapter, after a brief introduction to bioreactors (basic concepts, configurations, functions, and influencing factors), the applications of the bioreactor in microbial biomass, microbial biofuel conversion, and microbial electrochemical systems are described. Importantly, the role and significance of the bioreactor in the bioenergy process are discussed to provide a better understanding of the use of bioreactors in managing microbial biomass and energy conversion.

Modulation and modeling of monoclonal antibody N-linked glycosylation in mammalian cell perfusion reactors.

Science.gov (United States)

Karst, Daniel J; Scibona, Ernesto; Serra, Elisa; Bielser, Jean-Marc; Souquet, Jonathan; Stettler, Matthieu; Broly, Hervé; Soos, Miroslav; Morbidelli, Massimo; Villiger, Thomas K

2017-09-01

Mammalian cell perfusion cultures are gaining renewed interest as an alternative to traditional fed-batch processes for the production of therapeutic proteins, such as monoclonal antibodies (mAb). The steady state operation at high viable cell density allows the continuous delivery of antibody product with increased space-time yield and reduced in-process variability of critical product quality attributes (CQA). In particular, the production of a confined mAb N-linked glycosylation pattern has the potential to increase therapeutic efficacy and bioactivity. In this study, we show that accurate control of flow rates, media composition and cell density of a Chinese hamster ovary (CHO) cell perfusion bioreactor allowed the production of a constant glycosylation profile for over 20 days. Steady state was reached after an initial transition phase of 6 days required for the stabilization of extra- and intracellular processes. The possibility to modulate the glycosylation profile was further investigated in a Design of Experiment (DoE), at different viable cell density and media supplement concentrations. This strategy was implemented in a sequential screening approach, where various steady states were achieved sequentially during one culture. It was found that, whereas high ammonia levels reached at high viable cell densities (VCD) values inhibited the processing to complex glycan structures, the supplementation of either galactose, or manganese as well as their synergy significantly increased the proportion of complex forms. The obtained experimental data set was used to compare the reliability of a statistical response surface model (RSM) to a mechanistic model of N-linked glycosylation. The latter outperformed the response surface predictions with respect to its capability and reliability in predicting the system behavior (i.e., glycosylation pattern) outside the experimental space covered by the DoE design used for the model parameter estimation. Therefore, we can

Low-cost sensor system for non-invasive monitoring of cell growth in disposable bioreactors

OpenAIRE

Reinecke, Tobias; Biechele, Philipp; Schulte, V.; Scheper, Thomas; Zimmermann, Stefan

2015-01-01

To ensure productivity and product quality, the parameters of biotechnological processes need to be monitored. Along temperature or pH, one important parameter is the cell density in the culture medium. In this work, we present a low-cost sensor system for online cell growth monitoring in bioreactors via permittivity measurements based on coplanar transmission lines. To evaluate the sensor, E. coli cultivations are performed. We found a good correlation between optical density of the culture ...

Application of semifluidized bed bioreactor as novel bioreactor ...

African Journals Online (AJOL)

The conventional bioreactors such as pond digester, anaerobic filtration, up-flow anaerobic sludge blanket (UASB), up-flow anaerobic sludge fixed-film (UASFF), continuous stirred tank reactor (CSTR), anaerobic contact digestion and fluidized bed, used over the past decades are largely operated anaerobically. They have ...

SUBMERGED PERFUSION BIOREACTOR

DEFF Research Database (Denmark)

2010-01-01

NOVELTY - A biological device comprises a body (1) delimited by a rim (7) comprising recessed portion(s) that is a cavity in the rim of the body, and a first wall delimiting the recessed portion along the cavity. The recessed portion(s) comprise a first outlet orifice allowing the liquid medium t...

A Good Neighborhood for Cells: Bioreactor Demonstration System (BDS-05)

Science.gov (United States)

Chung, Leland W. K.; Goodwin, Thomas J. (Technical Monitor)

2002-01-01

Good neighborhoods help you grow. As with a city, the lives of a cell are governed by its neighborhood connections Connections that do not work are implicated in a range of diseases. One of those connections - between prostate cancer and bone cells - will be studied on STS-107 using the Bioreactor Demonstration System (BDS-05). To improve the prospects for finding novel therapies, and to identify biomarkers that predict disease progression, scientists need tissue models that behave the same as metastatic or spreading cancer. This is one of several NASA-sponsored lines of cell science research that use the microgravity environment of orbit in an attempt to grow lifelike tissue models for health research. As cells replicate, they "self associate" to form a complex matrix of collagens, proteins, fibers, and other structures. This highly evolved microenvironment tells each cell who is next door, how it should grow arid into what shapes, and how to respond to bacteria, wounds, and other stimuli. Studying these mechanisms outside the body is difficult because cells do not easily self-associate outside a natural environment. Most cell cultures produce thin, flat specimens that offer limited insight into how cells work together. Ironically, growing cell cultures in the microgravity of space produces cell assemblies that more closely resemble what is found in bodies on Earth. NASA's Bioreactor comprises a miniature life support system and a rotating vessel containing cell specimens in a nutrient medium. Orbital BDS experiments that cultured colon and prostate cancers have been highly promising.

40 CFR 258.41 - Project XL Bioreactor Landfill Projects.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Project XL Bioreactor Landfill... WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS Design Criteria § 258.41 Project XL Bioreactor Landfill Projects. (a) Buncombe County, North Carolina Project XL Bioreactor Landfill Requirements...

Glyco-engineering for biopharmaceutical production in moss bioreactors

Directory of Open Access Journals (Sweden)

Eva L. Decker

2014-07-01

Full Text Available The production of recombinant biopharmaceuticals (pharmaceutical proteins is a strongly growing area in the pharmaceutical industry. While most products to date are produced in mammalian cell cultures, namely CHO cells, plant-based production systems gained increasing acceptance over the last years. Different plant systems have been established which are suitable for standardization and precise control of cultivation conditions, thus meeting the criteria for pharmaceutical production.The majority of biopharmaceuticals comprise glycoproteins. Therefore, differences in protein glycosylation between humans and plants have to be taken into account and plant-specific glycosylation has to be eliminated to avoid adverse effects on quality, safety and efficacy of the products.The basal land plant Physcomitrella patens (moss has been employed for the recombinant production of high-value therapeutic target proteins (e.g., Vascular Endothelial Growth Factor, Complement Factor H, monoclonal antibodies, Erythropoietin. Being genetically excellently characterized and exceptionally amenable for precise gene targeting via homologous recombination, essential steps for the optimization of moss as a bioreactor for the production of recombinant proteins have been undertaken.Here, we discuss the glyco-engineering approaches to avoid non-human N- and O-glycosylation on target proteins produced in moss bioreactors.

An improvement of surfactin production by B. subtilis BBG131 using design of experiments in microbioreactors and continuous process in bubbleless membrane bioreactor.

Science.gov (United States)

Motta Dos Santos, Luiz Fernando; Coutte, François; Ravallec, Rozenn; Dhulster, Pascal; Tournier-Couturier, Lucie; Jacques, Philippe

2016-10-01

Culture medium elements were analysed by a screening DoE to identify their influence in surfactin specific production by a surfactin constitutive overproducing Bacillus subtilis strain. Statistics pointed the major enhancement caused by high glutamic acid concentrations, as well as a minor positive influence of tryptophan and glucose. Successively, a central composite design was performed in microplate bioreactors using a BioLector®, in which variations of these impressive parameters, glucose, glutamic acid and tryptophan concentrations were selected for optimization of product-biomass yield (YP/X). Results were exploited in combination with a RSM. In absolute terms, experiments attained an YP/X 3.28-fold higher than those obtained in Landy medium, a usual culture medium used for lipopeptide production by B. subtilis. Therefore, two medium compositions for enhancing biomass and surfactin specific production were proposed and tested in continuous regime in a bubbleless membrane bioreactor. An YP/X increase of 2.26-fold was observed in bioreactor scale. Copyright © 2016 Elsevier Ltd. All rights reserved.

Tubular membrane bioreactors for biotechnological processes.

Science.gov (United States)

Wolff, Christoph; Beutel, Sascha; Scheper, Thomas

2013-02-01

This article is an overview of bioreactors using tubular membranes such as hollow fibers or ceramic capillaries for cultivation processes. This diverse group of bioreactor is described here in regard to the membrane materials used, operational modes, and configurations. The typical advantages of this kind of system such as environments with low shear stress together with high cell densities and also disadvantages like poor oxygen supply are summed up. As the usage of tubular membrane bioreactors is not restricted to a certain organism, a brief overview of various applications covering nearly all types of cells from prokaryotic to eukaryotic cells is also given here.

Effect of sudden addition of PCE and bioreactor coupling to ZVI filters on performance of fluidized bed bioreactors operated in simultaneous electron acceptor modes.

Science.gov (United States)

Moreno-Medina, C U; Poggi-Varaldo, Hector M; Breton-Deval, L; Rinderknecht-Seijas, N

2017-11-01

The present work evaluated the effects of (i) feeding a water contaminated with 80 mg/L PCE to bioreactors seeded with inoculum not acclimated to PCE, (ii) coupling ZVI side filters to bioreactors, and (iii) working in different biological regimes, i.e., simultaneous methanogenic aeration and simultaneous methanogenic-denitrifying regimes, on fluidized bed bioreactor performance. Simultaneous electron acceptors refer to the simultaneous presence of two compounds operating as final electron acceptors in the biological respiratory chain (e.g., use of either O 2 or NO 3 - in combination with a methanogenic environment) in a bioreactor or environmental niche. Four lab-scale, mesophilic, fluidized bed bioreactors (bioreactors) were implemented. Two bioreactors were operated as simultaneous methanogenic-denitrifying (MD) units, whereas the other two were operated in partially aerated methanogenic (PAM) mode. In the first period, all bioreactors received a wastewater with 1 g chemical oxygen demand of methanol per liter (COD-methanol/L). In a second period, all the bioreactors received the wastewater plus 80 mg perchloroethylene (PCE)/L; at the start of period 2, one MD and one PAM were coupled to side sand-zero valent iron filters (ZVI). All bioreactors were inoculated with a microbial consortium not acclimated to PCE. In this work, the performance of the full period 1 and the first 60 days of period 2 is reported and discussed. The COD removal efficiency and the nitrate removal efficiency of the bioreactors essentially did not change between period 1 and period 2, i.e., upon PCE addition. On the contrary, specific methanogenic activity in PAM bioreactors (both with and without coupled ZVI filter) significantly decreased. This was consistent with a sharp fall of methane productivity in those bioreactors in period 2. During period 2, PCE removals in the range 86 to 97 % were generally observed; the highest removal corresponded to PAM bioreactors along with the

In vitro propagation of Stevia rebaudina plants using multiple shoot culture.

Science.gov (United States)

Nepovím, A; Vanek, T

1998-12-01

A multiple shoot culture was induced from nodal segments on MS medium containing half concentration of macroelements, 1% sucrose, and supplemented with NAA (0.01 mg/l). A bioreactor with hormone-free MS medium (300 ml) was inoculated with 1.5 g of the multiple shoot culture and cultivated for a month. The cultivating process of the multiple shoot culture in the bioreactor and the transfer into ex vitro conditions took about 8-9 weeks and produced approx. 600 new seedlings, that could be transferred from greenhouse to field conditions.

Effect of operating conditions in production of diagnostic Salmonella Enteritidis O-antigen-specific monoclonal antibody in different bioreactor systems.

Science.gov (United States)

Ayyildiz-Tamis, Duygu; Nalbantsoy, Ayse; Elibol, Murat; Deliloglu-Gurhan, Saime Ismet

2014-01-01

In this study, different cultivation systems such as roller bottles (RB), 5-L stirred-tank bioreactor (STR), and disposable bioreactors were used to cultivate hybridoma for lab-scale production of Salmonella Enteritidis O-antigen-specific monoclonal antibody (MAb). Hybridoma cell line was cultivated in either serum-containing or serum-free medium (SFM) culture conditions. In STR, MAb production scaled up to 4 L, and production capabilities of the cells were also evaluated in different featured production systems. Moreover, the growth parameters of the cells in all production systems such as glucose consumption, lactate and ammonia production, and also MAb productivities were determined. Collected supernatants from the reactors were concentrated by a cross-flow filtration system. In conclusion, cells were not adapted to SFM in RB and STR. Therefore, less MAb titer in both STR and RB systems with SFM was observed compared to the cultures containing fetal bovine serum-supplemented medium. A higher MAb titer was gained in the membrane-aerated system compared to those in STR and RB. Although the highest MAb titer was obtained in the static membrane bioreactor system, the highest productivity was obtained in STR operated in semicontinuous mode with overlay aeration.

Evaluation of a New Temporary Immersion Bioreactor System for Micropropagation of Cultivars of Eucalyptus, Birch and Fir

Directory of Open Access Journals (Sweden)

Edward Businge

2017-06-01

Full Text Available The use of liquid instead of solid culture medium for the micropropagation of plants offers advantages such as better access to medium components and scalability through possible automation of the processes. The objective of this work was to compare a new temporary immersion bioreactor (TIB to solid medium culture for the micropropagation of a selection of tree species micropropagated for commercial use: Nordmann fir (Abies nordmanniana (Steven Spach, Eucalyptus (E. grandis x E. urophylla, Downy birch (Betula pubescens Ehrh, and Curly birch (Betula pendula var. carelica. Cultivation of explants in the TIB resulted in a significant increase of multiplication rate and fresh weight of Eucalyptus and B. pendula, but not Betula pubescens. In addition, the fresh weight of embryogenic tissue and the maturation frequency of somatic embryos increased significantly when an embryogenic cell line of A. nordmanniana was cultivated in the TIB compared to solid culture medium. These results demonstrate the potential for scaling up and automating micropropagation by shoot multiplication and somatic embryogenesis in commercial tree species using a temporary immersion bioreactor.

Effects of flow configuration on bone tissue engineering using human mesenchymal stem cells in 3D chitosan composite scaffolds.

Science.gov (United States)

Sellgren, Katelyn L; Ma, Teng

2015-08-01

Perfusion bioreactor plays important role in supporting 3D bone construct development. Scaffolds of chitosan composites have been studied to support bone tissue regeneration from osteogenic progenitor cells including human mesenchymal stem cells (hMSC). In this study, porous scaffolds of hydroxyapatite (H), chitosan (C), and gelatin (G) were fabricated by phase-separation and press-fitted in the perfusion bioreactor system where media flow is configured either parallel or transverse with respect to the scaffolds to investigate the impact of flow configuration on hMSC proliferation and osteogenic differentiation. The in vitro results showed that the interstitial flow in the transverse flow (TF) constructs reduced cell growth during the first week of culture but improved spatial cell distribution and early onset of osteogenic differentiation measured by alkaline phosphatase and expression of osteogenic genes. After 14 days of bioreactor culture, the TF constructs have comparable cell number but higher expression of bone markers genes and proteins compared to the parallel flow constructs. To evaluate ectopic bone formation, the HCG constructs seeded with hMSCs pre-cultured under two flow configurations for 7 days were implanted in CD-1 nude mice. While Masson's Trichrom staining revealed bone formation in both constructs, the TF constructs have improved spatial cell and osteoid distribution throughout the 2.0 mm constructs. The results highlight the divergent effects of media flow over the course of construct development and suggest that the flow configuration is an important parameter regulating the cellular events leading to bone construct formation in the HCG scaffolds. © 2014 Wiley Periodicals, Inc.

Quantitative lung perfusion evaluation using Fourier decomposition perfusion MRI.

Science.gov (United States)

Kjørstad, Åsmund; Corteville, Dominique M R; Fischer, Andre; Henzler, Thomas; Schmid-Bindert, Gerald; Zöllner, Frank G; Schad, Lothar R

2014-08-01

To quantitatively evaluate lung perfusion using Fourier decomposition perfusion MRI. The Fourier decomposition (FD) method is a noninvasive method for assessing ventilation- and perfusion-related information in the lungs, where the perfusion maps in particular have shown promise for clinical use. However, the perfusion maps are nonquantitative and dimensionless, making follow-ups and direct comparisons between patients difficult. We present an approach to obtain physically meaningful and quantifiable perfusion maps using the FD method. The standard FD perfusion images are quantified by comparing the partially blood-filled pixels in the lung parenchyma with the fully blood-filled pixels in the aorta. The percentage of blood in a pixel is then combined with the temporal information, yielding quantitative blood flow values. The values of 10 healthy volunteers are compared with SEEPAGE measurements which have shown high consistency with dynamic contrast enhanced-MRI. All pulmonary blood flow (PBF) values are within the expected range. The two methods are in good agreement (mean difference = 0.2 mL/min/100 mL, mean absolute difference = 11 mL/min/100 mL, mean PBF-FD = 150 mL/min/100 mL, mean PBF-SEEPAGE = 151 mL/min/100 mL). The Bland-Altman plot shows a good spread of values, indicating no systematic bias between the methods. Quantitative lung perfusion can be obtained using the Fourier Decomposition method combined with a small amount of postprocessing. Copyright © 2013 Wiley Periodicals, Inc.

Finite element study of scaffold architecture design and culture conditions for tissue engineering.

Science.gov (United States)

Olivares, Andy L; Marsal, Elia; Planell, Josep A; Lacroix, Damien

2009-10-01

Tissue engineering scaffolds provide temporary mechanical support for tissue regeneration and transfer global mechanical load to mechanical stimuli to cells through its architecture. In this study the interactions between scaffold pore morphology, mechanical stimuli developed at the cell microscopic level, and culture conditions applied at the macroscopic scale are studied on two regular scaffold structures. Gyroid and hexagonal scaffolds of 55% and 70% porosity were modeled in a finite element analysis and were submitted to an inlet fluid flow or compressive strain. A mechanoregulation theory based on scaffold shear strain and fluid shear stress was applied for determining the influence of each structures on the mechanical stimuli on initial conditions. Results indicate that the distribution of shear stress induced by fluid perfusion is very dependent on pore distribution within the scaffold. Gyroid architectures provide a better accessibility of the fluid than hexagonal structures. Based on the mechanoregulation theory, the differentiation process in these structures was more sensitive to inlet fluid flow than axial strain of the scaffold. This study provides a computational approach to determine the mechanical stimuli at the cellular level when cells are cultured in a bioreactor and to relate mechanical stimuli with cell differentiation.

Anaerobic membrane bioreactor under extreme conditions (poster)

NARCIS (Netherlands)

Munoz Sierra, J.D.; De Kreuk, M.K.; Spanjers, H.; Van Lier, J.B.

2013-01-01

Membrane bioreactors ensure biomass retention by the application of micro or ultrafiltration processes. This allows operation at high sludge concentrations. Previous studies have shown that anaerobic membrane bioreactors is an efficient way to retain specialist microorganisms for treating

Bioinspired methodology for preparing magnetic responsive chitosan beads to be integrated in a tubular bioreactor for biomedical applications.

Science.gov (United States)

Song, Wenlong; Oliveira, Mariana B; Sher, Praveen; Gil, Sara; Nóbrega, J Miguel; Mano, João F

2013-08-01

Magnetic responsive chitosan beads were prepared using a methodology inspired by the rolling of water droplets over lotus leaves. Liquid precursors containing chitosan and magnetic microparticles were dispensed in the form of spherical droplets and crosslinked with genipin over synthetic superhydrophobic surfaces. Scanning electronic microscopy, histology and micro-computed tomography were employed to characterize the structure of the prepared composite beads and the inner distribution of the magnetic particles. Cellular metabolic activity tests showed that fibroblasts-like (L929 cell line) can adhere and proliferate on the prepared chitosan beads. We hypothesize that such spherical biomaterials could be integrated in a new concept of tubular bioreactor. The magnetic beads can be immobilized by an external magnetic field at specific positions and may be transported along the bioreactor by the drag of the culture medium flow. The system behavior was also studied through numerical modeling, which allowed to identify the relative importance of the main parameters, and to conclude that the distance between carrier beads plays a major role on their interaction with the culture medium and, consequently, on the overall system performance. In an up-scaled version of this bioreactor, the herein presented system may comprise different chambers in serial or parallel configurations. This constitutes a simple way of preparing magnetic responsive beads combined with a new design of bioreactor, which may find application in biomedicine and biotechnology, including in cell expansion for tissue engineering or for the production of therapeutic proteins to be used in cell therapies.

Bioinspired methodology for preparing magnetic responsive chitosan beads to be integrated in a tubular bioreactor for biomedical applications

International Nuclear Information System (INIS)

Song, Wenlong; Oliveira, Mariana B; Sher, Praveen; Gil, Sara; Mano, João F; Nóbrega, J Miguel

2013-01-01

Magnetic responsive chitosan beads were prepared using a methodology inspired by the rolling of water droplets over lotus leaves. Liquid precursors containing chitosan and magnetic microparticles were dispensed in the form of spherical droplets and crosslinked with genipin over synthetic superhydrophobic surfaces. Scanning electronic microscopy, histology and micro-computed tomography were employed to characterize the structure of the prepared composite beads and the inner distribution of the magnetic particles. Cellular metabolic activity tests showed that fibroblasts-like (L929 cell line) can adhere and proliferate on the prepared chitosan beads. We hypothesize that such spherical biomaterials could be integrated in a new concept of tubular bioreactor. The magnetic beads can be immobilized by an external magnetic field at specific positions and may be transported along the bioreactor by the drag of the culture medium flow. The system behavior was also studied through numerical modeling, which allowed to identify the relative importance of the main parameters, and to conclude that the distance between carrier beads plays a major role on their interaction with the culture medium and, consequently, on the overall system performance. In an up-scaled version of this bioreactor, the herein presented system may comprise different chambers in serial or parallel configurations. This constitutes a simple way of preparing magnetic responsive beads combined with a new design of bioreactor, which may find application in biomedicine and biotechnology, including in cell expansion for tissue engineering or for the production of therapeutic proteins to be used in cell therapies. (paper)

Renal perfusion scintiscan

Science.gov (United States)

... Radionuclide renal perfusion scan; Perfusion scintiscan - renal; Scintiscan - renal perfusion Images Kidney anatomy Kidney - blood and urine flow Intravenous pyelogram References Rottenberg G, Andi AC. Renal ...

Microfluidic engineered high cell density three-dimensional neural cultures

Science.gov (United States)

Cullen, D. Kacy; Vukasinovic, Jelena; Glezer, Ari; La Placa, Michelle C.

2007-06-01

Three-dimensional (3D) neural cultures with cells distributed throughout a thick, bioactive protein scaffold may better represent neurobiological phenomena than planar correlates lacking matrix support. Neural cells in vivo interact within a complex, multicellular environment with tightly coupled 3D cell-cell/cell-matrix interactions; however, thick 3D neural cultures at cell densities approaching that of brain rapidly decay, presumably due to diffusion limited interstitial mass transport. To address this issue, we have developed a novel perfusion platform that utilizes forced intercellular convection to enhance mass transport. First, we demonstrated that in thick (>500 µm) 3D neural cultures supported by passive diffusion, cell densities =104 cells mm-3), continuous medium perfusion at 2.0-11.0 µL min-1 improved viability compared to non-perfused cultures (p death and matrix degradation. In perfused cultures, survival was dependent on proximity to the perfusion source at 2.00-6.25 µL min-1 (p 90% viability in both neuronal cultures and neuronal-astrocytic co-cultures. This work demonstrates the utility of forced interstitial convection in improving the survival of high cell density 3D engineered neural constructs and may aid in the development of novel tissue-engineered systems reconstituting 3D cell-cell/cell-matrix interactions.

Operation of a fluidized-bed bioreactor for denitrification

International Nuclear Information System (INIS)

Hancher, C.W.; Taylor, P.A.; Napier, J.M.

1978-01-01

Two denitrification fluidized-bed bioreactors of the same length (i.e., 5 m) but with different inside diameters (i.e., 5 and 10 cm) have been operated on feed ranging in nitrate concentration from 200 to 2000 g/m 3 ; thus far, good agreement has been obtained. Two 10-cm-ID bioreactors operating in series have also been tested; the results are in accordance with predicted results based on the performance of a 5-cm-ID bioreactor. The overall denitrification rate in the dual 10-cm-ID bioreactor system was found to be 23 kg N(NO 3 - )/day-m 3 using feed with a nitrate concentration of 1800 g/m 3 . Data obtained in operating-temperature tests indicate that the maximum denitrification rate is achieved between 22 and 30 0 C. These data will form the basis of the design of our mobile pilot plant which consists of dual 20-cm-ID by 7.3-m-long bioreactors

In vitro culture of functionally active buffalo hepatocytes isolated by using a simplified manual perfusion method.

Directory of Open Access Journals (Sweden)

Santanu Panda

Full Text Available In farm animals, there is no suitable cell line available to understand liver-specific functions. This has limited our understanding of liver function and metabolism in farm animals. Culturing and maintenance of functionally active hepatocytes is difficult, since they survive no more than few days. Establishing primary culture of hepatocytes can help in studying cellular metabolism, drug toxicity, hepatocyte specific gene function and regulation. Here we provide a simple in vitro method for isolation and short-term culture of functionally active buffalo hepatocytes.Buffalo hepatocytes were isolated from caudate lobes by using manual enzymatic perfusion and mechanical disruption of liver tissue. Hepatocyte yield was (5.3 ± 0.66×107 cells per gram of liver tissue with a viability of 82.3 ± 3.5%. Freshly isolated hepatocytes were spherical with well contrasted border. After 24 hours of seeding onto fibroblast feeder layer and different extracellular matrices like dry collagen, matrigel and sandwich collagen coated plates, hepatocytes formed confluent monolayer with frequent clusters. Cultured hepatocytes exhibited typical cuboidal and polygonal shape with restored cellular polarity. Cells expressed hepatocyte-specific marker genes or proteins like albumin, hepatocyte nuclear factor 4α, glucose-6-phosphatase, tyrosine aminotransferase, cytochromes, cytokeratin and α1-antitrypsin. Hepatocytes could be immunostained with anti-cytokeratins, anti-albumin and anti α1-antitrypsin antibodies. Abundant lipid droplets were detected in the cytosol of hepatocytes using oil red stain. In vitro cultured hepatocytes could be grown for five days and maintained for up to nine days on buffalo skin fibroblast feeder layer. Cultured hepatocytes were viable for functional studies.We developed a convenient and cost effective technique for hepatocytes isolation for short-term culture that exhibited morphological and functional characteristics of active hepatocytes

Development of an Autonomous, Dual Chamber Bioreactor for the Growth of 3-Dimensional Epithelial-Stromal Tissues in Microgravity

Science.gov (United States)

Patel, Zarana S.; Wettergreen, Matthew A.; Huff, Janice L.

2014-01-01

We are developing a novel, autonomous bioreactor that can provide for the growth and maintenance in microgravity of 3-D organotypic epithelial-stromal cultures that require an air-liquid interface. These complex 3-D tissue models accurately represent the morphological features, differentiation markers, and growth characteristics observed in normal human epithelial tissues, including the skin, esophagus, lung, breast, pancreas, and colon. However, because of their precise and complex culture requirements, including that of an air-liquid interface, these 3-D models have yet to be utilized for life sciences research aboard the International Space Station. The development of a bioreactor for these cultures will provide the capability to perform biological research on the ISS using these realistic, tissue-like human epithelial-stromal cell models and will contribute significantly to advances in fundamental space biology research on questions regarding microgravity effects on normal tissue development, aging, cancer, and other disease processes. It will also allow for the study of how combined stressors, such as microgravity with radiation and nutritional deficiencies, affect multiple biological processes and will provide a platform for conducting countermeasure investigations on the ISS without the use of animal models. The technology will be autonomous and consist of a cell culture chamber that provides for air-liquid, liquid-liquid, and liquid-air exchanges within the chambers while maintaining the growth and development of the biological samples. The bioreactor will support multiple tissue types and its modular design will provide for incorporation of add-on capabilities such as microfluidics drug delivery, media sampling, and in situ biomarker analysis. Preliminary flight testing of the hardware will be conducted on a parabolic platform through NASA's Flight Opportunities Program.

An Update to Space Biomedical Research: Tissue Engineering in Microgravity Bioreactors

Directory of Open Access Journals (Sweden)

Abolfazl Barzegari

2012-03-01

Full Text Available Introduction: The severe need for constructing replacement tissues in organ transplantation has necessitated the development of tissue engineering approaches and bioreactors that can bring these approaches to reality. The inherent limitations of conventional bioreactors in generating realistic tissue constructs led to the devise of the microgravity tissue engineering that uses Rotating Wall Vessel (RWV bioreactors initially developed by NASA. Methods: In this review article, we intend to highlight some major advances and accomplishments in the rapidly-growing field of tissue engineering that could not be achieved without using microgravity. Results: Research is now focused on assembly of 3 dimensional (3D tissue fragments from various cell types in human body such as chondrocytes, osteoblasts, embryonic and mesenchymal stem cells, hepatocytes and pancreas islet cells. Hepatocytes cultured under microgravity are now being used in extracorporeal bioartificial liver devices. Tissue constructs can be used not only in organ replacement therapy, but also in pharmaco-toxicology and food safety assessment. 3D models of various cancers may be used in studying cancer development and biology or in high-throughput screening of anticancer drug candidates. Finally, 3D heterogeneous assemblies from cancer/immune cells provide models for immunotherapy of cancer. Conclusion: Tissue engineering in (simulated microgravity has been one of the stunning impacts of space research on biomedical sciences and their applications on earth.

Effects of hydraulic retention time on anaerobic hydrogenation performance and microbial ecology of bioreactors fed with glucose-peptone and starch-peptone

Energy Technology Data Exchange (ETDEWEB)

Li, Shiue-Lin; Chao, Yu-Chieh; Wang, Yu-Hsuan; Hsiao, Chia-Jung; Bai, Ming-Der [Department of Environmental Engineering, National Cheng-Kung University, No. 1, University Road, Tainan 701 (China); Whang, Liang-Ming; Wang, Yung-Fu; Cheng, Sheng-Shung [Department of Environmental Engineering, National Cheng-Kung University, No. 1, University Road, Tainan 701 (China); Sustainable Environment Research Center (SERC), National Cheng-Kung University, No. 1, University Road, Tainan 701 (China); Tseng, I.-Cheng [Sustainable Environment Research Center (SERC), National Cheng-Kung University, No. 1, University Road, Tainan 701 (China); Department of Life Science, National Cheng-Kung University, No. 1, University Road, Tainan 701 (China)

2010-01-15

This study evaluated anaerobic hydrogenation performance and microbial ecology in bioreactors operated at different hydraulic retention time (HRT) conditions and fed with glucose-peptone (GP) and starch-peptone (SP). The maximum hydrogen production rates for GP- and SP-fed bioreactors were found to be 1247 and 412 mmol-H{sub 2}/L/d at HRT of 2 and 3 h, respectively. At HRT > 8 h, hydrogen consumption due to peptone fermentation could occur and thus reduced hydrogen yield from carbohydrate fermentation. Results of cloning/sequencing and denaturant gradient gel electrophoresis (DGGE) indicated that Clostridium sporogenes and Clostridium celerecrescens were dominant hydrogen-producing bacteria in the GP-fed bioreactor, presumably due to their capability on protein hydrolysis. In the SP-fed bioreactor, Lactobacillus plantarum, Propionispira arboris, and Clostridium butyricum were found to be dominant populations, but the presence of P. arboris at HRT > 3 h might be responsible for a lower hydrogen yield from starch fermentation. As a result, optimizing HRT operation for bioreactors was considered an important asset in order to minimize hydrogen-consuming activities and thus maximize net hydrogen production. The limitation of simple parameters such as butyrate to acetate ratio (B/A ratio) in predicting hydrogen production was recognized in this study for bioreactors fed with multiple substrates. It is suggested that microbial ecology analysis, in addition to chemical analysis, should be performed when complex substrates and mixed cultures are used in hydrogen-producing bioreactors. (author)

Analysis of glutathione in supernatants and lysates of a human proximal tubular cell line from perfusion culture upon intoxication with cadmium chloride by HPLC and LC-ESI-MS

NARCIS (Netherlands)

Hahn, Hans; Huck, Christian W; Rainer, Matthias; Najam-ul-Haq, Muhammad; Bakry, Rania; Abberger, Thomas; Jennings, Paul; Pfaller, Walter; Bonn, Günther K

A simple and highly effective reversed-phase (RP) high-performance liquid chromatography (HPLC) method is described for analysing glutathione (GSH) and glutathione disulfide (GSSG) in out-flowing supernatants and lysates of perfusion cell cultures of human kidney cells (HK-2 cells) continuously

A microfluidic chip with a U-shaped microstructure array for multicellular spheroid formation, culturing and analysis

International Nuclear Information System (INIS)

Fu, Chien-Yu; Chang, Hwan-You; Tseng, Sheng-Yang; Yang, Shih-Mo; Hsu, Long; Liu, Cheng-Hsien

2014-01-01

Multicellular spheroids (MCS), formed by self-assembly of single cells, are commonly used as a three-dimensional cell culture model to bridge the gap between in vitro monolayer culture and in vivo tissues. However, current methods for MCS generation and analysis still suffer drawbacks such as being labor-intensive and of poor controllability, and are not suitable for high-throughput applications. This study demonstrates a novel microfluidic chip to facilitate MCS formation, culturing and analysis. The chip contains an array of U-shaped microstructures fabricated by photopolymerizing the poly(ethylene glycol) diacrylate hydrogel through defining the ultraviolet light exposure pattern with a photomask. The geometry of the U-shaped microstructures allowed trapping cells into the pocket through the actions of fluid flow and the force of gravity. The hydrogel is non-adherent for cells, promoting the formation of MCS. Its permselective property also facilitates exchange of nutrients and waste for MCS, while providing protection of MCS from shearing stress during the medium perfusion. Heterotypic MCS can be formed easily by manipulating the cell trapping steps. Subsequent drug susceptibility analysis and long-term culture could also be achieved within the same chip. This MCS formation and culture platform can be used as a micro-scale bioreactor and applied in many cell biology and drug testing studies. (paper)

Online monitoring of cartilage tissue in a novel bioreactor

Science.gov (United States)

von der Burg, E.; von Buttlar, M.; Grill, W.

2011-04-01

Standard techniques for the analysis of biological tissues like immunohistochemical staining are typically invasive and lead to mortification of cells. Non-invasive monitoring is an important element of regenerative medicine because implants and components of implants should be 100% quality-checked with non-invasive and therefore also marker-free methods. We report on a new bioreactor for the production of collagen scaffolds seeded with Mesenchymal Stem Cells (MSCs). It contains a computer controlled mechanical activation and ultrasonic online monitoring and has been constructed for the in situ determination of ultrasonic and rheological parameters. During the cultivation period of about two weeks the scaffold is periodically compressed by two movable pistons for improved differentiation of the MSCs. This periodic compression beneficially ensures the supply with nutrition even inside the sample. During the physiological stimuli, rheological properties are measured by means of highly sensitive load cells. In addition measurements of the speed of sound in the sample and in the culture medium, with frequencies up to 16 MHz, are performed continuously. Therefore piezoceramic transducers are attached to the pistons and emit and detect ultrasonic waves, travelling through the pistons, the sample and the culture medium. The time-of-flight (TOF) of the ultrasonic signals is determined in real time with the aid of chirped excitation and correlation procedures with a resolution of at least 10 ps. The implemented ultrasonic measurement scheme allows beside the speed of sound measurements the detection of the distance between the pistons with a resolution better than 100 nm. The developed monitoring delivers information on rigidity, fluid dynamics and velocity of sound in the sample and in the culture medium. The hermetically sealed bioreactor with its life support system provides a biocompatible environment for MSCs for long time cultivation.

Tissue-engineered bone constructed in a bioreactor for repairing critical-sized bone defects in sheep.

Science.gov (United States)

Li, Deqiang; Li, Ming; Liu, Peilai; Zhang, Yuankai; Lu, Jianxi; Li, Jianmin

2014-11-01

Repair of bone defects, particularly critical-sized bone defects, is a considerable challenge in orthopaedics. Tissue-engineered bones provide an effective approach. However, previous studies mainly focused on the repair of bone defects in small animals. For better clinical application, repairing critical-sized bone defects in large animals must be studied. This study investigated the effect of a tissue-engineered bone for repairing critical-sized bone defect in sheep. A tissue-engineered bone was constructed by culturing bone marrow mesenchymal-stem-cell-derived osteoblast cells seeded in a porous β-tricalcium phosphate ceramic (β-TCP) scaffold in a perfusion bioreactor. A critical-sized bone defect in sheep was repaired with the tissue-engineered bone. At the eighth and 16th week after the implantation of the tissue-engineered bone, X-ray examination and histological analysis were performed to evaluate the defect. The bone defect with only the β-TCP scaffold served as the control. X-ray showed that the bone defect was successfully repaired 16 weeks after implantation of the tissue-engineered bone; histological sections showed that a sufficient volume of new bones formed in β-TCP 16 weeks after implantation. Eight and 16 weeks after implantation, the volume of new bones that formed in the tissue-engineered bone group was more than that in the β-TCP scaffold group (P bone improved osteogenesis in vivo and enhanced the ability to repair critical-sized bone defects in large animals.

Effect of growth conditions on the biodegradation kinetics of toluene by P. putida 54G in a vapor phase bioreactor

International Nuclear Information System (INIS)

Mirpuri, R.; Jones, W.; Krieger, E.; McFeters, G.

1994-01-01

Biodegradation of volatile organic compounds such as petroleum hydrocarbons and xenobiotic agents in the vapor phase is a promising new concept in well-head and end-of-pipe treatment which may have wide application where in-situ approaches are not feasible. The microbial degradation of the volatile organics can be carried out in vapor phase bioreactors which contain inert packing materials. Scale-up of these reactors from a bench scale to a pilot plant can best be achieved by the use of a predictive model, the success of which depends on accurate estimates of parameters defined in the model such as biodegradation kinetic and stoichiometric coefficients. The phenomena of hydrocarbon stress and injury may also affect performance of a vapor phase bioreactor. Batch kinetic studies on the biodegradation of toluene by P. Putida 54G will be compared to those obtained from continuous culture studies for both suspended and biofilm cultures of the same microorganism. These results will be compared to the activity of the P. putida 54G biofilm in a vapor phase bioreactor to evaluate the impact of hydrocarbon stress and injury on biodegradative processes

Scale-up of bioreactors: The concept of bioreactor number and its relation to the physiology of industrial microorganisms at different scales

Energy Technology Data Exchange (ETDEWEB)

De Ford, D

1988-01-01

The objective of this research is to provide a novel approach to the problem of scale-up of fermentations. The work subscribes the idea that two regions appear in bioreactors as the volume increases. The first is where high oxygen transfer occurs and the second is where low oxygen transfer occurs. It is assumed that organisms grown in a stirred tank fermenter travel in a cyclical manner through these two regions. A dimensionless factor is developed, the bioreactor number. Using this number the performance of any stirred tank fermenter can be described as a function of its geometry, operating conditions and physical properties of media. A mathematical model for the prediction of the physiological response of aerobic micro-organisms (specific growth rate, final cell concentration and product synthesis) as a function of the bioreactor number is also developed. It was adjusted by using the results of fermentations performed in a specially designed experimental rig allowing the simulation of fermenters with various bioreactor numbers. If the bioreactor and physiological models are linked it is possible to predict how micro-organisms respond when geometry, operating conditions or media properties are changed in a bioreactor. This approach is a tool for decision making in the design and operation of fermenters.

Determination of Ammonia Oxidizing Bacteria and Nitrate Oxidizing Bacteria in Wastewater and Bioreactors

Science.gov (United States)

Francis, Somilez Asya

2014-01-01

The process of water purification has many different physical, chemical, and biological processes. One part of the biological process is the task of ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB). Both play critical roles in the treatment of wastewater by oxidizing toxic compounds. The broad term is nitrification, a naturally occurring process that is carried out by AOB and NOB by using oxidation to convert ammonia to nitrite and nitrite to nitrate. To monitor this biological activity, bacterial staining was performed on wastewater contained in inoculum tanks and biofilm samples from bioreactors. Using microscopy and qPCR, the purpose of this experiment was to determine if the population of AOB and NOB in wastewater and membrane bioreactors changed depending on temperature and hibernation conditions to determine the optimal parameters for AOB/NOB culture to effectively clean wastewater.

Enrichment of carbon monoxide utilising microorganisms from methanogenic bioreactor sludge

OpenAIRE

Pereira, Ana Luísa; Stams, Alfons Johannes Maria; Alves, M. M.; Sousa, D. Z.

2015-01-01

Conversion of CO is the rate limiting step during anaerobic conversion of syngas (a gaseous mixture mainly composed of CO, CO2 and H2). In this work we study the microbial diversity in anaerobic sludge submitted to extended contact to syngas in a multi-orifice baffled bioreactor (MOBB). Methane was the main product resulting from syngas conversion in the MOBB. Enrichment cultures started with this sludge produced methane as final product, but also acetate. 16S rRNA gene analysis revealed a pr...

Structure-Activity Relationships of Bioengineered Heparin/Heparan Sulfates Produced in Different Bioreactors

Directory of Open Access Journals (Sweden)

Ha Na Kim

2017-05-01

Full Text Available Heparin and heparan sulfate are structurally-related carbohydrates with therapeutic applications in anticoagulation, drug delivery, and regenerative medicine. This study explored the effect of different bioreactor conditions on the production of heparin/heparan sulfate chains via the recombinant expression of serglycin in mammalian cells. Tissue culture flasks and continuously-stirred tank reactors promoted the production of serglycin decorated with heparin/heparan sulfate, as well as chondroitin sulfate, while the serglycin secreted by cells in the tissue culture flasks produced more highly-sulfated heparin/heparan sulfate chains. The serglycin produced in tissue culture flasks was effective in binding and signaling fibroblast growth factor 2, indicating the utility of this molecule in drug delivery and regenerative medicine applications in addition to its well-known anticoagulant activity.

Comparison of membrane bioreactor technology and conventional ...

African Journals Online (AJOL)

The purpose of this paper was to review the use of membrane bioreactor technology as an alternative for treating the discharged effluent from a bleached kraft mill by comparing and contrasting membrane bioreactors with conventional activated sludge systems for wastewater treatment. There are many water shortage ...

An innovative membrane bioreactor for methane biohydroxylation.

Science.gov (United States)

Pen, N; Soussan, L; Belleville, M-P; Sanchez, J; Charmette, C; Paolucci-Jeanjean, D

2014-12-01

In this study, a membrane bioreactor (MBR) was developed for efficient, safe microbial methane hydroxylation with Methylosinus trichosporium OB3b. This innovative MBR, which couples a bioreactor with two gas/liquid macroporous membrane contactors supplying the two gaseous substrates (methane and oxygen) was operated in fed-batch mode. The feasibility and the reproducibility of this new biohydroxylation process were first demonstrated. The mass transfer within this MBR was twice that observed in a batch reactor in similar conditions. The productivity reached with this MBR was 75±25mgmethanol(gdrycell)(-1)h(-1). Compared to the literature, this value is 35times higher than that obtained with the only other fed-batch membrane bioreactor reported, which was run with dense membranes, and is comparable to those obtained with bioreactors fed by bubble-spargers. However, in the latter case, an explosive gas mixture can be formed, a problem that is avoided with the MBR. Copyright © 2014 Elsevier Ltd. All rights reserved.

Rotating three-dimensional dynamic culture of adult human bone marrow-derived cells for tissue engineering of hyaline cartilage.

Science.gov (United States)

Sakai, Shinsuke; Mishima, Hajime; Ishii, Tomoo; Akaogi, Hiroshi; Yoshioka, Tomokazu; Ohyabu, Yoshimi; Chang, Fei; Ochiai, Naoyuki; Uemura, Toshimasa

2009-04-01

The method of constructing cartilage tissue from bone marrow-derived cells in vitro is considered a valuable technique for hyaline cartilage regenerative medicine. Using a rotating wall vessel (RWV) bioreactor developed in a NASA space experiment, we attempted to efficiently construct hyaline cartilage tissue from human bone marrow-derived cells without using a scaffold. Bone marrow aspirates were obtained from the iliac crest of nine patients during orthopedic operation. After their proliferation in monolayer culture, the adherent cells were cultured in the RWV bioreactor with chondrogenic medium for 2 weeks. Cells from the same source were cultured in pellet culture as controls. Histological and immunohistological evaluations (collagen type I and II) and quantification of glycosaminoglycan were performed on formed tissues and compared. The engineered constructs obtained using the RWV bioreactor showed strong features of hyaline cartilage in terms of their morphology as determined by histological and immunohistological evaluations. The glycosaminoglycan contents per microg DNA of the tissues were 10.01 +/- 3.49 microg/microg DNA in the case of the RWV bioreactor and 6.27 +/- 3.41 microg/microg DNA in the case of the pellet culture, and their difference was significant. The RWV bioreactor could provide an excellent environment for three-dimensional cartilage tissue architecture that can promote the chondrogenic differentiation of adult human bone marrow-derived cells.

Bioreactor Design for Tendon/Ligament Engineering

OpenAIRE

Wang, Tao; Gardiner, Bruce S.; Lin, Zhen; Rubenson, Jonas; Kirk, Thomas B.; Wang, Allan; Xu, Jiake; Smith, David W.; Lloyd, David G.; Zheng, Ming H.

2012-01-01

Tendon and ligament injury is a worldwide health problem, but the treatment options remain limited. Tendon and ligament engineering might provide an alternative tissue source for the surgical replacement of injured tendon. A bioreactor provides a controllable environment enabling the systematic study of specific biological, biochemical, and biomechanical requirements to design and manufacture engineered tendon/ligament tissue. Furthermore, the tendon/ligament bioreactor system can provide a s...

Biodegradation of phenolic waste liquors in stirred-tank, packed-bed, and fluidized-bed bioreactors

Energy Technology Data Exchange (ETDEWEB)

Holladay, D W; Hancher, G W; Chilcote, D D; Scott, C D

1978-11-01

The biological degradation of phenolic scrub liquors similar to those that arise in coal conversion processes was studied for symbiotic bacterial populations contained in a continuously stirred tank bioreactor, a three-phase packed-bed bioreactor, and a three-phase, fluidized-bed bioreactor. The conversions of phenol compounds were comparable in the three-phase, packed-bed bioreactor and the continuously stirred tank bioreactor; however, the packed-bed bioreactor degradation rates were as much as twice those in the continuously stirred tank bioreactor, and packed-bed bioreactor retention times were as low as one- tenth those of the continuously stirred tank bioreactors (minimum time was 12 hours).

Evaluation of mechanical and morphologic features of PLLA membranes as supports for perfusion cells culture systems

Energy Technology Data Exchange (ETDEWEB)

Montesanto, S., E-mail: salvatore.montesanto1985@gmail.com [Department of Civil, Environmental, Aerospace, Materials Engineering (DICAM), University of Palermo, Viale delle Scienze Ed. 8, 90128 Palermo (Italy); Brucato, V. [Department of Civil, Environmental, Aerospace, Materials Engineering (DICAM), University of Palermo, Viale delle Scienze Ed. 8, 90128 Palermo (Italy); La Carrubba, V. [Department of Civil, Environmental, Aerospace, Materials Engineering (DICAM), University of Palermo, Viale delle Scienze Ed. 8, 90128 Palermo (Italy); Euro-Mediterranean Institute of Science and Technology (IEMEST), Via Michele Miraglia, 20, 90128 Palermo (Italy)

2016-12-01

Porous biodegradable PLLA membranes, which can be used as supports for perfusion cell culture systems were designed, developed and characterized. PLLA membranes were prepared via diffusion induced phase separation (DIPS). A glass slab was coated with a binary PLLA–dioxane solution (8 wt.% PLLA) via dip coating, then pool immersed in two subsequent coagulation baths, and finally dried in a humidity-controlled environment. Surface and mechanical properties were evaluated by measuring pore size, porosity via scanning electron microscopy, storage modulus, loss modulus and loss angle by using a dynamic mechanical analysis (DMA). Cell adhesion assays on different membrane surfaces were also performed by using a standard count method. Results provide new insights into the foaming methods for producing polymeric membranes and supply indications on how to optimise the fabrication parameters to design membranes for tissue cultures and regeneration. - Highlights: • Design, development and characterization of porous biodegradable PLLA membranes via DIPS technology. • Easy-to-tune processing parameters in terms of surface and volumetric properties and cell adhesion. • Evaluation of the impact of the interconnection degree on membrane's mechanical properties. • Evaluation of cell adhesion on different membrane surface textures.

Evaluation of mechanical and morphologic features of PLLA membranes as supports for perfusion cells culture systems

International Nuclear Information System (INIS)

Montesanto, S.; Brucato, V.; La Carrubba, V.

2016-01-01

Porous biodegradable PLLA membranes, which can be used as supports for perfusion cell culture systems were designed, developed and characterized. PLLA membranes were prepared via diffusion induced phase separation (DIPS). A glass slab was coated with a binary PLLA–dioxane solution (8 wt.% PLLA) via dip coating, then pool immersed in two subsequent coagulation baths, and finally dried in a humidity-controlled environment. Surface and mechanical properties were evaluated by measuring pore size, porosity via scanning electron microscopy, storage modulus, loss modulus and loss angle by using a dynamic mechanical analysis (DMA). Cell adhesion assays on different membrane surfaces were also performed by using a standard count method. Results provide new insights into the foaming methods for producing polymeric membranes and supply indications on how to optimise the fabrication parameters to design membranes for tissue cultures and regeneration. - Highlights: • Design, development and characterization of porous biodegradable PLLA membranes via DIPS technology. • Easy-to-tune processing parameters in terms of surface and volumetric properties and cell adhesion. • Evaluation of the impact of the interconnection degree on membrane's mechanical properties. • Evaluation of cell adhesion on different membrane surface textures.

Correlation between mass transfer coefficient kLa and relevant operating parameters in cylindrical disposable shaken bioreactors on a bench-to-pilot scale.

Science.gov (United States)

Klöckner, Wolf; Gacem, Riad; Anderlei, Tibor; Raven, Nicole; Schillberg, Stefan; Lattermann, Clemens; Büchs, Jochen

2013-12-02

Among disposable bioreactor systems, cylindrical orbitally shaken bioreactors show important advantages. They provide a well-defined hydrodynamic flow combined with excellent mixing and oxygen transfer for mammalian and plant cell cultivations. Since there is no known universal correlation between the volumetric mass transfer coefficient for oxygen kLa and relevant operating parameters in such bioreactor systems, the aim of this current study is to experimentally determine a universal kLa correlation. A Respiration Activity Monitoring System (RAMOS) was used to measure kLa values in cylindrical disposable shaken bioreactors and Buckingham's π-Theorem was applied to define a dimensionless equation for kLa. In this way, a scale- and volume-independent kLa correlation was developed and validated in bioreactors with volumes from 2 L to 200 L. The final correlation was used to calculate cultivation parameters at different scales to allow a sufficient oxygen supply of tobacco BY-2 cell suspension cultures. The resulting equation can be universally applied to calculate the mass transfer coefficient for any of seven relevant cultivation parameters such as the reactor diameter, the shaking frequency, the filling volume, the viscosity, the oxygen diffusion coefficient, the gravitational acceleration or the shaking diameter within an accuracy range of +/- 30%. To our knowledge, this is the first kLa correlation that has been defined and validated for the cited bioreactor system on a bench-to-pilot scale.

Impact of Furfural on Rapid Ethanol Production Using a Membrane Bioreactor

Directory of Open Access Journals (Sweden)

Mohammad J. Taherzadeh

2013-03-01

Full Text Available A membrane bioreactor was developed to counteract the inhibition effect of furfural in ethanol production. Furfural, a major inhibitor in lignocellulosic hydrolyzates, is a highly toxic substance which is formed from pentose sugars released during the acidic degradation of lignocellulosic materials. Continuous cultivations with complete cell retention were performed at a high dilution rate of 0.5 h−1. Furfural was added directly into the bioreactor by pulse injection or by addition into the feed medium to obtain furfural concentrations ranging from 0.1 to 21.8 g L−1. At all pulse injections of furfural, the yeast was able to convert the furfural very rapidly by in situ detoxification. When injecting 21.8 g L−1 furfural to the cultivation, the yeast converted it by a specific conversion rate of 0.35 g g−1 h−1. At high cell density, Saccharomyces cerevisiae could tolerate very high furfural levels without major changes in the ethanol production. During the continuous cultures when up to 17.0 g L−1 furfural was added to the inlet medium, the yeast successfully produced ethanol, whereas an increase of furfural to 18.6 and 20.6 g L−1 resulted in a rapidly decreasing ethanol production and accumulation of sugars in the permeate. This study show that continuous ethanol fermentations by total cell retention in a membrane bioreactor has a high furfural tolerance and can conduct rapid in situ detoxification of medium containing high furfural concentrations.

Large-scale production of lentiviral vector in a closed system hollow fiber bioreactor

Directory of Open Access Journals (Sweden)

Jonathan Sheu

Full Text Available Lentiviral vectors are widely used in the field of gene therapy as an effective method for permanent gene delivery. While current methods of producing small scale vector batches for research purposes depend largely on culture flasks, the emergence and popularity of lentiviral vectors in translational, preclinical and clinical research has demanded their production on a much larger scale, a task that can be difficult to manage with the numbers of producer cell culture flasks required for large volumes of vector. To generate a large scale, partially closed system method for the manufacturing of clinical grade lentiviral vector suitable for the generation of induced pluripotent stem cells (iPSCs, we developed a method employing a hollow fiber bioreactor traditionally used for cell expansion. We have demonstrated the growth, transfection, and vector-producing capability of 293T producer cells in this system. Vector particle RNA titers after subsequent vector concentration yielded values comparable to lentiviral iPSC induction vector batches produced using traditional culture methods in 225 cm2 flasks (T225s and in 10-layer cell factories (CF10s, while yielding a volume nearly 145 times larger than the yield from a T225 flask and nearly three times larger than the yield from a CF10. Employing a closed system hollow fiber bioreactor for vector production offers the possibility of manufacturing large quantities of gene therapy vector while minimizing reagent usage, equipment footprint, and open system manipulation.

Systematic microcarrier screening and agitated culture conditions improves human mesenchymal stem cell yield in bioreactors

Science.gov (United States)

Rafiq, Qasim A.; Coopman, Karen; Nienow, Alvin W.

2016-01-01

Abstract Production of human mesenchymal stem cells for allogeneic cell therapies requires scalable, cost‐effective manufacturing processes. Microcarriers enable the culture of anchorage‐dependent cells in stirred‐tank bioreactors. However, no robust, transferable methodology for microcarrier selection exists, with studies providing little or no reason explaining why a microcarrier was employed. We systematically evaluated 13 microcarriers for human bone marrow‐derived MSC (hBM‐MSCs) expansion from three donors to establish a reproducible and transferable methodology for microcarrier selection. Monolayer studies demonstrated input cell line variability with respect to growth kinetics and metabolite flux. HBM‐MSC1 underwent more cumulative population doublings over three passages in comparison to hBM‐MSC2 and hBM‐MSC3. In 100 mL spinner flasks, agitated conditions were significantly better than static conditions, irrespective of donor, and relative microcarrier performance was identical where the same microcarriers outperformed others with respect to growth kinetics and metabolite flux. Relative growth kinetics between donor cells on the microcarriers were the same as the monolayer study. Plastic microcarriers were selected as the optimal microcarrier for hBM‐MSC expansion. HBM‐MSCs were successfully harvested and characterised, demonstrating hBM‐MSC immunophenotype and differentiation capacity. This approach provides a systematic method for microcarrier selection, and the findings identify potentially significant bioprocessing implications for microcarrier‐based allogeneic cell therapy manufacture. PMID:26632496

Modelling across bioreactor scales: methods, challenges and limitations

DEFF Research Database (Denmark)

Gernaey, Krist

that it is challenging and expensive to acquire experimental data of good quality that can be used for characterizing gradients occurring inside a large industrial scale bioreactor. But which model building methods are available? And how can one ensure that the parameters in such a model are properly estimated? And what......Scale-up and scale-down of bioreactors are very important in industrial biotechnology, especially with the currently available knowledge on the occurrence of gradients in industrial-scale bioreactors. Moreover, it becomes increasingly appealing to model such industrial scale systems, considering...

New bioreactor for in situ simultaneous measurement of bioluminescence and cell density

Science.gov (United States)

Picart, Pascal; Bendriaa, Loubna; Daniel, Philippe; Horry, Habib; Durand, Marie-José; Jouvanneau, Laurent; Thouand, Gérald

2004-03-01

This article presents a new device devoted to the simultaneous measurement of bioluminescence and optical density of a bioluminescent bacterial culture. It features an optoelectronic bioreactor with a fully autoclavable module, in which the bioluminescent bacteria are cultivated, a modulated laser diode dedicated to optical density measurement, and a detection head for the acquisition of both bioluminescence and optical density signals. Light is detected through a bifurcated fiber bundle. This setup allows the simultaneous estimation of the bioluminescence and the cell density of the culture medium without any sampling. The bioluminescence is measured through a highly sensitive photomultiplier unit which has been photometrically calibrated to allow light flux measurements. This was achieved by considering the bioluminescence spectrum and the full optical transmission of the device. The instrument makes it possible to measure a very weak light flux of only a few pW. The optical density is determined through the laser diode and a photodiode using numerical synchronous detection which is based on the power spectrum density of the recorded signal. The detection was calibrated to measure optical density up to 2.5. The device was validated using the Vibrio fischeri bacterium which was cultivated under continuous culture conditions. A very good correlation between manual and automatic measurements processed with this instrument has been demonstrated. Furthermore, the optoelectronic bioreactor enables determination of the luminance of the bioluminescent bacteria which is estimated to be 6×10-5 W sr-1 m-2 for optical density=0.3. Experimental results are presented and discussed.

Nitrate Removal Rates in Denitrifying Bioreactors During Storm Flows

Science.gov (United States)

Pluer, W.; Walter, T.

2017-12-01

Field denitrifying bioreactors are designed to reduce excess nitrate (NO3-) pollution in runoff from agricultural fields. Field bioreactors saturate organic matter to create conditions that facilitate microbial denitrification. Prior studies using steady flow in lab-scale bioreactors showed that a hydraulic retention time (HRT) between 4 and 10 hours was optimal for reducing NO3- loads. However, during storm-induced events, flow rate and actual HRT fluctuate. These fluctuations have the potential to disrupt the system in significant ways that are not captured by the idealized steady-flow HRT models. The goal of this study was to investigate removal rate during dynamic storm flows of variable rates and durations. Our results indicate that storm peak flow and duration were not significant controlling variables. Instead, we found high correlations (p=0.004) in average removal rates between bioreactors displaying a predominantly uniform flow pattern compared with bioreactors that exhibited preferential flow (24.4 and 21.4 g N m-3 d-1, respectively). This suggests that the internal flow patterns are a more significant driver of removal rate than external factors of the storm hydrograph. Designing for flow patterns in addition to theoretical HRT will facilitate complete mixing within the bioreactors. This will help maximize excess NO3- removal during large storm-induced runoff events.

Design and validation of a dynamic cell-culture system for bone biology research and exogenous tissue-engineering applications.

Science.gov (United States)

Allori, Alexander C; Davidson, Edward H; Reformat, Derek D; Sailon, Alexander M; Freeman, James; Vaughan, Adam; Wootton, David; Clark, Elizabeth; Ricci, John L; Warren, Stephen M

2016-10-01

Bone lacunocanalicular fluid flow ensures chemotransportation and provides a mechanical stimulus to cells. Traditional static cell-culture methods are ill-suited to study the intricacies of bone biology because they ignore the three-dimensionality of meaningful cellular networks and the lacunocanalicular system; furthermore, reliance on diffusion alone for nutrient supply and waste product removal effectively limits scaffolds to 2-3 mm thickness. In this project, a flow-perfusion system was custom-designed to overcome these limitations: eight adaptable chambers housed cylindrical cell-seeded scaffolds measuring 12 or 24 mm in diameter and 1-10 mm in thickness. The porous scaffolds were manufactured using a three-dimensional (3D) periodic microprinting process and were composed of hydroxyapatite/tricalcium phosphate with variable thicknesses, strut sizes, pore sizes and structural configurations. A multi-channel peristaltic pump drew medium from parallel reservoirs and perfused it through each scaffold at a programmable rate. Hermetically sealed valves permitted sampling or replacement of medium. A gas-permeable membrane allowed for gas exchange. Tubing was selected to withstand continuous perfusion for > 2 months without leakage. Computational modelling was performed to assess the adequacy of oxygen supply and the range of fluid shear stress in the bioreactor-scaffold system, using 12 × 6 mm scaffolds, and these models suggested scaffold design modifications that improved oxygen delivery while enhancing physiological shear stress. This system may prove useful in studying complex 3D bone biology and in developing strategies for engineering thick 3D bone constructs. Copyright © 2013 John Wiley & Sons, Ltd. Copyright © 2013 John Wiley & Sons, Ltd.

Processing highly porous calcium phosphate ceramics for use in bioreactor cores for culturing human liver cells in-vitro

Science.gov (United States)

Finoli, Anthony

Chronic liver disease is the 11th highest cause of death in the United States claiming over 30,000 lives in 2009. The current treatment for chronic liver failure is liver transplantation but the availability of tissue is far less than the number of patients in need. To develop human liver tissue in the lab a 3D culturing environment must be created to support the growth of a complex tissue. Hydroxyapatite (HAp) has been chosen as a scaffold material because of its biocompatibility in the body and the ability to create a bioresorbable scaffold. By using a ceramic material, it is possible to create a three dimensional, protective environment in which tissue can grow. The first part of this study is to examine the behavior of adult human liver cells grown on composites of HAp and different biocompatible hydrogels. Porous HAp has been created using an emulsion foaming technique and cells are injected into the structure after being suspended in a hydrogel and are kept in culture for up to 28 days. Functional assays, gene expression and fluorescent microscopy will be used to examine these cultures. The second part of this study will be to develop a processing technique to create a resorbable scaffold that incorporates a vascular system template. Previous experiments have shown the high temperature decomposition of HAp into resorbable calcium phosphates will be used to create a multiphase material. By controlling the amount of transformation product formed, it is proposed that the resorption of the scaffold can be tailored. To introduce a pore network to guide the growth of a vascular system, a positive-negative casting technique has also been developed. A positive polymer copy can be made of a natural vascular system and ceramic is foamed around the copy. During sintering, the polymer is pyrolyzed leaving a multiscale pore network in the ceramic. By combining these techniques, it is proposed that a calcium phosphate bioreactor core can be processed that is suitable for

Modeling of a membrane bioreactor for production of biodiesel

International Nuclear Information System (INIS)

Solano, Paola Andrea; Moncada, Jorge Andres; Cardona, Carlos Ariel; Ruiz, Orlando Simon

2008-01-01

Through the use of an enzymatic catalyst lipase, produced by Candida Antarctica a membrane bioreactor was modeled and simulated to obtain biodiesel from palm oil and ethanol. A conversion of 0.97 was reached for a residence time of 10.64 min. The membrane bioreactor was compared to a CSTR reactor, where a conversion of 0.76 was obtained. It was concluded that the membrane bioreactor is a better way of producing biodiesel than the CSTR

Cascades of bioreactors

NARCIS (Netherlands)

Gooijer, de C.D.

1995-01-01

In this thesis a common phenomenon in bioprocess engineering is described : the execution of a certain bioprocess in more than one bioreactor. Chapter 1, a review, classifies bioprocesses by means of a number of characteristics :
i) processes with a variable

Fundamentals of membrane bioreactors materials, systems and membrane fouling

CERN Document Server

Ladewig, Bradley

2017-01-01

This book provides a critical, carefully researched, up-to-date summary of membranes for membrane bioreactors. It presents a comprehensive and self-contained outline of the fundamentals of membrane bioreactors, especially their relevance as an advanced water treatment technology. This outline helps to bring the technology to the readers’ attention, and positions the critical topic of membrane fouling as one of the key impediments to its more widescale adoption. The target readership includes researchers and industrial practitioners with an interest in membrane bioreactors.

A bioartificial environment for kidney epithelial cells based on a supramolecular polymer basement membrane mimic and an organotypical culture system.

Science.gov (United States)

Mollet, Björne B; Bogaerts, Iven L J; van Almen, Geert C; Dankers, Patricia Y W

2017-06-01

Renal applications in healthcare, such as renal replacement therapies and nephrotoxicity tests, could potentially benefit from bioartificial kidney membranes with fully differentiated and functional human tubular epithelial cells. A replacement of the natural environment of these cells is required to maintain and study cell functionality cell differentiation in vitro. Our approach was based on synthetic supramolecular biomaterials to mimic the natural basement membrane (BM) on which these cells grow and a bioreactor to provide the desired organotypical culture parameters. The BM mimics were constructed from ureidopyrimidinone (UPy)-functionalized polymer and bioactive peptides by electrospinning. The resultant membranes were shown to have a hierarchical fibrous BM-like structure consisting of self-assembled nanofibres within the electrospun microfibres. Human kidney-2 (HK-2) epithelial cells were cultured on the BM mimics under organotypical conditions in a custom-built bioreactor. The bioreactor facilitated in situ monitoring and functionality testing of the cultures. Cell viability and the integrity of the epithelial cell barrier were demonstrated inside the bioreactor by microscopy and transmembrane leakage of fluorescently labelled inulin, respectively. Furthermore, HK-2 cells maintained a polarized cell layer and showed modulation of both gene expression of membrane transporter proteins and metabolic activity of brush border enzymes when subjected to a continuous flow of culture medium inside the new bioreactor for 21 days. These results demonstrated that both the culture and study of renal epithelial cells was facilitated by the bioartificial in vitro environment that is formed by synthetic supramolecular BM mimics in our custom-built bioreactor. Copyright © 2015 John Wiley & Sons, Ltd. Copyright © 2015 John Wiley & Sons, Ltd.

Perfusion abnormalities in congenital and neoplastic pulmonary disease: comparison of MR perfusion and multislice CT imaging

International Nuclear Information System (INIS)

Boll, Daniel T.; Lewin, Jonathan S.; Young, Philip; Gilkeson, Robert C.; Siwik, Ernest S.

2005-01-01

The aim of this work was to assess magnetic resonance (MR) perfusion patterns of chronic, nonembolic pulmonary diseases of congenital and neoplastic origin and to compare the findings with results obtained with pulmonary, contrast-enhanced multislice computed tomography (CT) imaging to prove that congenital and neoplastic pulmonary conditions require MR imaging over the pulmonary perfusion cycle to successfully and directly detect changes in lung perfusion patterns. Twenty-five patients underwent concurrent CT and MR evaluation of chronic pulmonary diseases of congenital (n=15) or neoplastic (n=10) origin. Analysis of MR perfusion and contrast-enhanced CT datasets was realized by defining pulmonary and vascular regions of interest in corresponding positions. MR perfusion calculated time-to-peak enhancement, maximal enhancement and the area under the perfusion curve. CT datasets provided pulmonary signal-to-noise ratio measurements. Vessel centerlines of bronchial arteries were determined. Underlying perfusion type, such as pulmonary arterial or systemic arterial supply, as well as regions with significant variations in perfusion were determined statistically. Analysis of the pulmonary perfusion pattern detected pulmonary arterial supply in 19 patients; six patients showed systemic arterial supply. In pulmonary arterial perfusion, MR and multislice CT imaging consistently detected the perfusion type and regions with altered perfusion patterns. In bronchial arterial supply, MR perfusion and CT imaging showed significant perfusion differences. Patients with bronchial arterial supply had bronchial arteries ranging from 2.0 to 3.6 mm compared with submillimeter diameters in pulmonary arterial perfusion. Dynamic MR imaging of congenital and neoplastic pulmonary conditions allowed characterization of the pulmonary perfusion type. CT imaging suggested the presence of systemic arterial perfusion by visualizing hypertrophied bronchial arteries. (orig.)

Isolation and characterization of two novel alkalitolerant sulfidogens from a Thiopaq bioreactor, Desulfonatronum alkalitolerans sp. nov., and Sulfurospirillum alkalitolerans sp. nov.

NARCIS (Netherlands)

Sorokin, D.Y.; Tourova, T.P.; Muyzer, G.

2013-01-01

wo obligately anaerobic sulfidogenic bacterial strains were isolated from the full-scale Thiopaq bioreactor in Lelystad (The Netherlands) removing H2S from biogas under oxygen-limiting and moderately haloalkaline conditions. Strain HSRB-L represents a dominant culturable sulfate-reducing bacterium

Estimation of the Maximum Theoretical Productivity of Fed-Batch Bioreactors

Energy Technology Data Exchange (ETDEWEB)

Bomble, Yannick J [National Renewable Energy Laboratory (NREL), Golden, CO (United States); St. John, Peter C [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Crowley, Michael F [National Renewable Energy Laboratory (NREL), Golden, CO (United States)

2017-10-18

A key step towards the development of an integrated biorefinery is the screening of economically viable processes, which depends sharply on the yields and productivities that can be achieved by an engineered microorganism. In this study, we extend an earlier method which used dynamic optimization to find the maximum theoretical productivity of batch cultures to explicitly include fed-batch bioreactors. In addition to optimizing the intracellular distribution of metabolites between cell growth and product formation, we calculate the optimal control trajectory of feed rate versus time. We further analyze how sensitive the productivity is to substrate uptake and growth parameters.

Immobilized yeast in bioreactor for alcohol fermentation

International Nuclear Information System (INIS)

Handy, M.K.; Kim, K.

1986-01-01

Mutant of Saccharomyces cerevisiae was developed using a Co-60 source. Cells were immobilized onto sterile, channeled alumina beads and packed into bioreactor column under controlled temperature. Feedstocks containing substrate and nutrients were fed into the bioreactor at specific rates. Beads with greatest porosity and surface area produced the most ethanol. Factors affecting ethanol productivity included: temperature, pH, flow rate, nutrients and substrate in the feedstock

Efficient Production Process for Food Grade Acetic Acid by Acetobacter aceti in Shake Flask and in Bioreactor Cultures

Directory of Open Access Journals (Sweden)

Hassan M. Awad

2012-01-01

Full Text Available Acetic acid is one of the important weak acids which had long history in chemical industries. This weak organic acid has been widely used as one of the key intermediate for many chemical, detergent, wood and food industries. The production of this acid is mainly carried out using submerged fermentation system and the standard strain Acetobacter aceti. In the present work, six different media were chosen from the literatures and tested for acetic acid production. The highest acetic acid production was produced in medium composed of glucose, yeast extract and peptone. The composition of this medium was optimized by changing the concentration of medium components. The optimized medium was composed of (g/L: glucose, 100; yeast extract, 12 and peptone 5 and yielded 53 g/L acetic acid in shake flask after 144 h fermentation. Further optimization in the production process was achieved by transferring the process to semi-industrial scale 16-L stirred tank bioreactor and cultivation under controlled pH condition. Under fully aerobic conditions, the production of acetic acid reached maximal concentration of about 76 g/L and 51 g/L for uncontrolled and controlled pH cultures, respectively.

Construction of a Simple Multipurpose Airlift Bioreactor and its ...

African Journals Online (AJOL)

BSN

The aim of the present research is to develop a simple airlift bioreactor which can be operated even ... compression metal. The bioreactor is mixed ... the method developed by (Bailey and Olis, .... (Ed) Concise Encyclopedia of Bio-resources.

A bone tissue engineering strategy based on starch scaffolds and bone marrow cells cultured in a flow perfusion bioreactor

Science.gov (United States)

Gomes, Maria Manuela Estima

tecido osseo, uma vez que estas celulas podem ser facilmente recolhidas do proprio paciente a tratar por metodos nao-invasivos (bioppsia) e em quantidades suficientes. Alem disso, tratando-se de uma fonte de celulas autologas (obtidas do proprio paciente) permitem evitar os riscos de transmissao de doencas contagiosas e/ou de rejeicao pelo sistema imunologico. • Estudo da influencia das condicoes de cultura in vitro geradas por um bioreactor de perfusao (em comparacao com os metodos tradicionais de cultura em condicoes estaticas) no desenvolvimento dos materiais hibridos, compostos pelas celulas e scaffolds, assim como as interaccoes do ambiente proporcionado por este sistema de cultura com as diferentes estruturas/arquitecturas e porosidades dos scaffolds utilizados. Estes objectivos convergem para o objectivo geral desta tese que consistiu no desenvolvimento de uma terapia de engenharia do tecido osseo alternativa as existentes e com potencial para vir a ser posteriormente utilizada na pratica clinica. Este objectivo foi avaliado atraves do estudo da funcionalidade dos materiais hibridos obtidos em diferentes condicoes de cultura in vitro (e utilizando diferentes scaffolds), partindo do principio que o sistema de perfusao poderia eventualmente superar as limitacoes de difusao tipicas dos sistema de cultura estatica e simultaneamente proporcionar estimulos mecânicos as celulas, semelhantes aos encontrados em condicoes fisiologicas. (Abstract shortened by ProQuest.).

Cyclic mechanical stimulation rescues achilles tendon from degeneration in a bioreactor system.

Science.gov (United States)

Wang, Tao; Lin, Zhen; Ni, Ming; Thien, Christine; Day, Robert E; Gardiner, Bruce; Rubenson, Jonas; Kirk, Thomas B; Smith, David W; Wang, Allan; Lloyd, David G; Wang, Yan; Zheng, Qiujian; Zheng, Ming H

2015-12-01

Physiotherapy is one of the effective treatments for tendinopathy, whereby symptoms are relieved by changing the biomechanical environment of the pathological tendon. However, the underlying mechanism remains unclear. In this study, we first established a model of progressive tendinopathy-like degeneration in the rabbit Achilles. Following ex vivo loading deprivation culture in a bioreactor system for 6 and 12 days, tendons exhibited progressive degenerative changes, abnormal collagen type III production, increased cell apoptosis, and weakened mechanical properties. When intervention was applied at day 7 for another 6 days by using cyclic tensile mechanical stimulation (6% strain, 0.25 Hz, 8 h/day) in a bioreactor, the pathological changes and mechanical properties were almost restored to levels seen in healthy tendon. Our results indicated that a proper biomechanical environment was able to rescue early-stage pathological changes by increased collagen type I production, decreased collagen degradation and cell apoptosis. The ex vivo model developed in this study allows systematic study on the effect of mechanical stimulation on tendon biology. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

Laser doppler perfusion imaging

International Nuclear Information System (INIS)

Waardell, K.

1992-01-01

Recording of tissue perfusion is important in assessing the influence of peripheral vascular diseases on the microcirculation. This thesis reports on a laser doppler perfusion imager based on dynamic light scattering in tissue. When a low power He-Ne laser beam sequentally scans the tissue, moving blood cells generate doppler components in the back-scattered light. A fraction of this light is detected by a photodetector and converted into an electrical signal. In the processor, a signal proportional to the tissue perfusion at each measurement site is calculated and stored. When the scanning procedure is completed, a color-coded perfusion image is presented on a monitor. To convert important aspects of the perfusion image into more quantitative parameters, data analysis functions are implemented in the software. A theory describing the dependence of the distance between individual measurement points and detector on the system amplification factor is proposed and correction algorithms are presented. The performance of the laser doppler perfusion imager was evaluated using a flow simulator. A linear relationship between processor output signal and flow through the simulator was demonstrated for blood cell concentrations below 0.2%. The median sampling depth of the laser beam was simulated by a Monte Carlo technique and estimated to 235 μm. The perfusion imager has been used in the clinic to study perfusion changes in port wine stains treated with argon laser and to investigate the intensity and extension of the cutaneous axon reflex response after electrical nerve stimulation. The fact that perfusion can be visualized without touching the tissue implies elimination of sterilization problems, thus simplifying clinical investigations of perfusion in association with diagnosis and treatment of peripheral vascular diseases. 22 refs

Neuropsychological Correlates of Brain Perfusion SPECT in Patients with Macrophagic Myofasciitis.

Directory of Open Access Journals (Sweden)

Axel Van Der Gucht

Full Text Available Patients with aluminum hydroxide adjuvant-induced macrophagic myofasciitis (MMF complain of arthromyalgias, chronic fatigue and cognitive deficits. This study aimed to characterize brain perfusion in these patients.Brain perfusion SPECT was performed in 76 consecutive patients (aged 49±10 y followed in the Garches-Necker-Mondor-Hendaye reference center for rare neuromuscular diseases. Images were acquired 30 min after intravenous injection of 925 MBq 99mTc-ethylcysteinate dimer (ECD at rest. All patients also underwent a comprehensive battery of neuropsychological tests, within 1.3±5.5 mo from SPECT. Statistical parametric maps (SPM12 were obtained for each test using linear regressions between each performance score and brain perfusion, with adjustment for age, sex, socio-cultural level and time delay between brain SPECT and neuropsychological testing.SPM analysis revealed positive correlation between neuropsychological scores (mostly exploring executive functions and brain perfusion in the posterior associative cortex, including cuneus/precuneus/occipital lingual areas, the periventricular white matter/corpus callosum, and the cerebellum, while negative correlation was found with amygdalo-hippocampal/entorhinal complexes. A positive correlation was also observed between brain perfusion and the posterior associative cortex when the time elapsed since last vaccine injection was investigated.Brain perfusion SPECT showed a pattern of cortical and subcortical changes in accordance with the MMF-associated cognitive disorder previously described. These results provide a neurobiological substrate for brain dysfunction in aluminum hydroxide adjuvant-induced MMF patients.

Organotypic lung culture: A new model for studying ischemia and ex vivo perfusion in lung transplantation.

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Baste, Jean-Marc; Gay, Arnaud; Smail, Hassiba; Noël, Romain; Bubenheim, Michael; Begueret, Hugues; Morin, Jean-Paul; Litzler, Pierre-Yves

2015-01-01

Donors after cardiac death (DCD) in lung transplantation is considered as a solution for organ shortage. However, it is characterized by warm ischemic period, which could be involved in severe Ischemia-Reperfusion lesion (IR) with early graft dysfunction. We describe a new hybrid model combining in vivo ischemia followed by in vitro reoxygenation using organ-specific culture. A hybrid model using in vivo ischemic period followed by in vitro lung slice reoxygenation was set up in rat to mimic DCD in lung transplantation with in vitro perfusion. Different markers (bioenergetics, oxidant stress assays, and histology) were measured to evaluate the viability of lung tissue after different ischemic times (I-0, I-1, I-2, I-4, I-15 hours) and reoxygenation times (R-0, R-1, R-4, R-24 hours). No differences were found in cell viability, ATP concentrations, extracellular LDH assays or histology, demonstrating extensive viability of up to 4 hours in lung tissue warm ischemia. We found oxidative stress mainly during the ischemic period with no burst at reoxygenation. Cytosolic anti-oxidant system was involved first (I-0,I-1,I-2) followed by mitochondrial anti-oxidant system for extensive ischemia (I-4). Histological features showed differences in this model of ischemia-reoxygenation between bronchial epithelium and lung parenchymal cells, with epithelium regeneration after 2 hours of warm ischemia and 24 hours of perfusion. The results of our hybrid model experiment suggest extensive lung viability of up to 4 hours ischemia. Our model could be an interesting tool to evaluate ex vivo reconditioning techniques after different in vivo lung insults.

Methanol utilizing Desulfotomaculum species utilizes hydrogen in a methanol-fed sulfate-reducing bioreactor.

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Balk, Melike; Weijma, Jan; Goorissen, Heleen P; Ronteltap, Mariska; Hansen, Theo A; Stams, Alfons J M

2007-01-01

A sulfate-reducing bacterium, strain WW1, was isolated from a thermophilic bioreactor operated at 65 degrees C with methanol as sole energy source in the presence of sulfate. Growth of strain WW1 on methanol or acetate was inhibited at a sulfide concentration of 200 mg l(-1), while on H2/CO2, no apparent inhibition occurred up to a concentration of 500 mg l(-1). When strain WW1 was co-cultured under the same conditions with the methanol-utilizing, non-sulfate-reducing bacteria, Thermotoga lettingae and Moorella mulderi, both originating from the same bioreactor, growth and sulfide formation were observed up to 430 mg l(-1). These results indicated that in the co-cultures, a major part of the electron flow was directed from methanol via H2/CO2 to the reduction of sulfate to sulfide. Besides methanol, acetate, and hydrogen, strain WW1 was also able to use formate, malate, fumarate, propionate, succinate, butyrate, ethanol, propanol, butanol, isobutanol, with concomitant reduction of sulfate to sulfide. In the absence of sulfate, strain WW1 grew only on pyruvate and lactate. On the basis of 16S rRNA analysis, strain WW1 was most closely related to Desulfotomaculum thermocisternum and Desulfotomaculum australicum. However, physiological properties of strain WW1 differed in some aspects from those of the two related bacteria.

Distribution and viability of fetal and adult human bone marrow stromal cells in a biaxial rotating vessel bioreactor after seeding on polymeric 3D additive manufactured scaffolds

Directory of Open Access Journals (Sweden)

Anne eLeferink

2015-10-01

Full Text Available One of the conventional approaches in tissue engineering is the use of scaffolds in combination with cells to obtain mechanically stable tissue constructs in vitro prior to implantation. Additive manufacturing by fused deposition modeling is a widely used technique to produce porous scaffolds with defined pore network, geometry, and therewith defined mechanical properties. Bone marrow derived mesenchymal stromal cells (MSCs are promising candidates for tissue engineering based cell therapies due to their multipotent character. One of the hurdles to overcome when combining additive manufactured scaffolds with MSCs is the resulting heterogeneous cell distribution and limited cell proliferation capacity. In this study, we show that the use of a biaxial rotating bioreactor, after static culture of human fetal MSCs (hfMSCs seeded on synthetic polymeric scaffolds, improved the homogeneity of cell and extracellular matrix (ECM distribution and increased the total cell number. Furthermore, we show that the relative mRNA expression levels of indicators for stemness and differentiation are not significantly changed upon this bioreactor culture, whereas static culture shows variations of several indicators for stemness and differentiation. The biaxial rotating bioreactor presented here offers a homogeneous distribution of hfMSCs, enabling studies on MSCs fate in additive manufactured scaffolds without inducing undesired differentiation.

Plantform Bioreactor for Mass Micropropagation of Date Palm.

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Almusawi, Abdulminam H A; Sayegh, Abdullah J; Alshanaw, Ansam M S; Griffis, John L

2017-01-01

A novel protocol for the commercial production of date palm through micropropagation is presented. This protocol includes the use of a semisolid medium alternation or in combination with a temporary immersion system (TIS, Plantform bioreactor) in date palm micropropagation. The use of the Plantform bioreactor for date palm results in an improved multiplication rate, reduced micropropagation time, and improved weaning success. It also reduces the cost of saleable units and thus improves economic return for commercial micropropagation. The use of the Plantform bioreactor successfully addresses other hindrances that can occur during the scale-up of date palm micropropagation, including asynchrony of somatic embryos, limited maturation of somatic embryos, and highly variable germination frequencies of embryos.

Pharmaceutical proteins produced in plant bioreactor in recent years ...

African Journals Online (AJOL)

Plant bioreactor, also called molecular farming, has enormous potential to produce recombinant proteins infinitely. Products expressed in plants have natural physico-chemical properties and bioactivities. Plant bioreactor could be a safe, economic and convenient production system, and can been widely applied in ...

Integrating biological treatment of crop residue into a hydroponic sweetpotato culture

Science.gov (United States)

Trotman, A. A.; David, P. P.; Bonsi, C. K.; Hill, W. A.; Mortley, D. G.; Loretan, P. A.

1997-01-01

Residual biomass from hydroponic culture of sweetpotato [Ipomoea batatas (L.) Lam.] was degraded using natural bacterial soil isolates. Sweetpotato was grown for 120 days in hydroponic culture with a nutrient solution comprised of a ratio of 80% modified half Hoagland solution to 20% filtered effluent from an aerobic starch hydrolysis bioreactor. The phytotoxicity of the effluent was assayed with `Waldmann's Green' lettuce (Lactuca sativa L.) and the ratio selected after a 60-day bioassay using sweetpotato plants propagated vegetatively from cuttings. Controlled environment chamber experiments were conducted to investigate the impact of filtrate from biological treatment of crop residue on growth and storage root production with plants grown in a modified half Hoagland solution. Incorporation of bioreactor effluent, reduced storage root yield of `Georgia Jet' sweetpotato but the decrease was not statistically significant when compared with yield for plants cultured in a modified half Hoagland solution without filtrate. However, yield of `TU-82-155' sweetpotato was significantly reduced when grown in a modified half Hoagland solution into which filtered effluent had been incorporated. Total biomass was significantly reduced for both sweetpotato cultivars when grown in bioreactor effluent. The leaf area and dry matter accumulation were significantly (P < 0.05) reduced for both cultivars when grown in solution culture containing 20% filtered effluent.

Computational modeling of adherent cell growth in a hollow-fiber membrane bioreactor for large-scale 3-D bone tissue engineering.

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Mohebbi-Kalhori, Davod; Behzadmehr, Amin; Doillon, Charles J; Hadjizadeh, Afra

2012-09-01

The use of hollow-fiber membrane bioreactors (HFMBs) has been proposed for three-dimensional bone tissue growth at the clinical scale. However, to achieve an efficient HFMB design, the relationship between cell growth and environmental conditions must be determined. Therefore, in this work, a dynamic double-porous media model was developed to determine nutrient-dependent cell growth for bone tissue formation in a HFMB. The whole hollow-fiber scaffold within the bioreactor was treated as a porous domain in this model. The domain consisted of two interpenetrating porous regions, including a porous lumen region available for fluid flow and a porous extracapillary space filled with a collagen gel that contained adherent cells for promoting long-term growth into tissue-like mass. The governing equations were solved numerically and the model was validated using previously published experimental results. The contributions of several bioreactor design and process parameters to the performance of the bioreactor were studied. The results demonstrated that the process and design parameters of the HFMB significantly affect nutrient transport and thus cell behavior over a long period of culture. The approach presented here can be applied to any cell type and used to develop tissue engineering hollow-fiber scaffolds.

Staying alive! Sensors used for monitoring cell health in bioreactors.

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O'Mara, P; Farrell, A; Bones, J; Twomey, K

2018-01-01

Current and next generation sensors such as pH, dissolved oxygen (dO) and temperature sensors that will help drive the use of single-use bioreactors in industry are reviewed. The current trend in bioreactor use is shifting from the traditional fixed bioreactors to the use of single-use bioreactors (SUBs). However as the shift in paradigm occurs there is now a greater need for sensor technology to play 'catch up' with the innovation of bioreactor technology. Many of the sensors still in use today rely on technology created in the 1960's such as the Clark-type dissolved oxygen sensor or glass pH electrodes. This is due to the strict requirements of sensors to monitor bioprocesses resulting in the use of traditional well understood methods, making it difficult to incorporate new sensor technology into industry. A number of advances in sensor technology have been achieved in recent years, a few of these advances and future research will also be discussed in this review. Copyright © 2017 Elsevier B.V. All rights reserved.

The effect of perfusion culture on proliferation and differentiation of human mesenchymal stem cells on biocorrodible bone replacement material

International Nuclear Information System (INIS)

Farack, J.; Wolf-Brandstetter, C.; Glorius, S.; Nies, B.; Standke, G.; Quadbeck, P.; Worch, H.; Scharnweber, D.

2011-01-01

Biocorrodible iron foams were coated with different calcium phosphate phases (CPP) to obtain a bioactive surface and controlled degradation. Further adhesion, proliferation and differentiation of SaOs-2 and human mesenchymal stem cells were investigated under both static and dynamic culture conditions. Hydroxyapatite (HA; [Ca 10 (PO 4 ) 6 OH 2 ]) coated foams released 500 μg/g iron per day for Dulbecco's modified eagle medium (DMEM) and 250 μg/g iron per day for McCoys, the unmodified reference 1000 μg/g iron per day for DMEM and 500 μg/g iron per day for McCoys, while no corrosion could be detected on brushite (CaHPO 4 ) coated foams. Using a perfusion culture system with conditions closer to the in vivo situation, cells proliferated and differentiated on iron foams coated with either brushite or HA while in static cell culture cells could proliferate only on Fe-brushite. We conclude that the degradation behaviour of biocorrodible iron foams can be varied by different calcium phosphate coatings, offering opportunities for design of novel bone implants. Further studies will focus on the influence of different modifications of iron foams on the expression of oxidative stress enzymes. Additional information about in vivo reactions and remodelling behaviour are expected from testing in implantation studies.

Abnormal perfusion on myocardial perfusion SPECT in patients with Wolff-Parkinson-White syndrome

International Nuclear Information System (INIS)

Kang, Do Young; Cha, Kwang Soo; Han, Seung Ho; Park, Tae Ho; Kim, Moo Hyun; Kim, Young Dae

2005-01-01

Abnormal myocardial perfusion may be caused by ventricular preexcitation, but its location, extent, severity and correlation with accessory pathway (AP) are not established. We evaluated perfusion patterns on myocardial perfusion SPECT and location of AP in patients with WPW (Wolff-Parkison-White) syndrome. Adenosine Tc-99m MIBI or Tl-201 myocardial perfusion SPECT was performed in 11 patients with WPW syndrome. Perfusion defects (PD) were compared to AP location based on ECT with Fitzpatrick's algorithm of electrophysiologic study and radiofrequency catheter ablation. Patients had atypical chest discomfort or no symptom. Risk of coronary artery disease (CAD) was below 0.1 in 11 patients using the nomogram to estimate the probability of CAD. Coronary angiography was performed in 4 patients(mid-LAD 50% in one, normal in others). In 4 patients, AP localization was done by electrophysiologic study and radiofrequency catheter ablation (RFCA). Small to large extent (11.0 ± 8.5%, range:3 ∼ 35%) and mild to moderate severity (-71 ± 42.7%, range:-217 ∼ -39%) of reversible (n=9) or fixed (n=1) perfusion defects were noted. One patients with right free wall (right lateral) AP showed normal. PD locations were variable following the location of AP. One patient with left lateral wall AP was followed 6 weeks after RFCA and showed significantly decreased PD on SPECT with successful ablation. Myocardial perfusion defect showed variable extent, severity and location in patients with WPW syndrome. Abnormal perfusion defect showed in most of all patients, but if did not seem to be correlated specifically with location of accessory pathway and coronary artery disease. Therefore myocardial perfusion SPECT should be interpreted carefully in patients with WPW syndrome

Abnormal perfusion on myocardial perfusion SPECT in patients with Wolff-Parkinson-White syndrome

Energy Technology Data Exchange (ETDEWEB)

Kang, Do Young; Cha, Kwang Soo; Han, Seung Ho; Park, Tae Ho; Kim, Moo Hyun; Kim, Young Dae [Donga University College of Medicine, Busan (Korea, Republic of)

2005-02-15

Abnormal myocardial perfusion may be caused by ventricular preexcitation, but its location, extent, severity and correlation with accessory pathway (AP) are not established. We evaluated perfusion patterns on myocardial perfusion SPECT and location of AP in patients with WPW (Wolff-Parkison-White) syndrome. Adenosine Tc-99m MIBI or Tl-201 myocardial perfusion SPECT was performed in 11 patients with WPW syndrome. Perfusion defects (PD) were compared to AP location based on ECT with Fitzpatrick's algorithm of electrophysiologic study and radiofrequency catheter ablation. Patients had atypical chest discomfort or no symptom. Risk of coronary artery disease (CAD) was below 0.1 in 11 patients using the nomogram to estimate the probability of CAD. Coronary angiography was performed in 4 patients(mid-LAD 50% in one, normal in others). In 4 patients, AP localization was done by electrophysiologic study and radiofrequency catheter ablation (RFCA). Small to large extent (11.0 {+-} 8.5%, range:3 {approx} 35%) and mild to moderate severity (-71 {+-} 42.7%, range:-217 {approx} -39%) of reversible (n=9) or fixed (n=1) perfusion defects were noted. One patients with right free wall (right lateral) AP showed normal. PD locations were variable following the location of AP. One patient with left lateral wall AP was followed 6 weeks after RFCA and showed significantly decreased PD on SPECT with successful ablation. Myocardial perfusion defect showed variable extent, severity and location in patients with WPW syndrome. Abnormal perfusion defect showed in most of all patients, but if did not seem to be correlated specifically with location of accessory pathway and coronary artery disease. Therefore myocardial perfusion SPECT should be interpreted carefully in patients with WPW syndrome.

Numerical simulation of fluid field and in vitro three-dimensional fabrication of tissue-engineered bones in a rotating bioreactor and in vivo implantation for repairing segmental bone defects.

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Song, Kedong; Wang, Hai; Zhang, Bowen; Lim, Mayasari; Liu, Yingchao; Liu, Tianqing

2013-03-01

In this paper, two-dimensional flow field simulation was conducted to determine shear stresses and velocity profiles for bone tissue engineering in a rotating wall vessel bioreactor (RWVB). In addition, in vitro three-dimensional fabrication of tissue-engineered bones was carried out in optimized bioreactor conditions, and in vivo implantation using fabricated bones was performed for segmental bone defects of Zelanian rabbits. The distribution of dynamic pressure, total pressure, shear stress, and velocity within the culture chamber was calculated for different scaffold locations. According to the simulation results, the dynamic pressure, velocity, and shear stress around the surface of cell-scaffold construction periodically changed at different locations of the RWVB, which could result in periodical stress stimulation for fabricated tissue constructs. However, overall shear stresses were relatively low, and the fluid velocities were uniform in the bioreactor. Our in vitro experiments showed that the number of cells cultured in the RWVB was five times higher than those cultured in a T-flask. The tissue-engineered bones grew very well in the RWVB. This study demonstrates that stress stimulation in an RWVB can be beneficial for cell/bio-derived bone constructs fabricated in an RWVB, with an application for repairing segmental bone defects.

Process cost and facility considerations in the selection of primary cell culture clarification technology.

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Felo, Michael; Christensen, Brandon; Higgins, John

2013-01-01

The bioreactor volume delineating the selection of primary clarification technology is not always easily defined. Development of a commercial scale process for the manufacture of therapeutic proteins requires scale-up from a few liters to thousands of liters. While the separation techniques used for protein purification are largely conserved across scales, the separation techniques for primary cell culture clarification vary with scale. Process models were developed to compare monoclonal antibody production costs using two cell culture clarification technologies. One process model was created for cell culture clarification by disc stack centrifugation with depth filtration. A second process model was created for clarification by multi-stage depth filtration. Analyses were performed to examine the influence of bioreactor volume, product titer, depth filter capacity, and facility utilization on overall operating costs. At bioreactor volumes 5,000 L, clarification using centrifugation followed by depth filtration offers significant cost savings. For bioreactor volumes of ∼ 2,000 L, clarification costs are similar between depth filtration and centrifugation. At this scale, factors including facility utilization, available capital, ease of process development, implementation timelines, and process performance characterization play an important role in clarification technology selection. In the case study presented, a multi-product facility selected multi-stage depth filtration for cell culture clarification at the 500 and 2,000 L scales of operation. Facility implementation timelines, process development activities, equipment commissioning and validation, scale-up effects, and process robustness are examined. © 2013 American Institute of Chemical Engineers.

Regeneration from embryogenic callus and suspension cultures of ...

African Journals Online (AJOL)

ehab

2012-04-25

Apr 25, 2012 ... Micropropagation, through seed culture has been studied in C. martinii ... Effect of 2,4-D on total and embryogenic callus production ... ethanol for 1 min, followed by immersion in 20% Clorox (NaOCl ... Three flasks from each treatment were cultured. ... shaker at 110 rpm. ...... Plants cultivated in bioreactor of.

The Use of an Acellular Oxygen Carrier in a Human Liver Model of Normothermic Machine Perfusion.

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Laing, Richard W; Bhogal, Ricky H; Wallace, Lorraine; Boteon, Yuri; Neil, Desley A H; Smith, Amanda; Stephenson, Barney T F; Schlegel, Andrea; Hübscher, Stefan G; Mirza, Darius F; Afford, Simon C; Mergental, Hynek

2017-11-01

Normothermic machine perfusion of the liver (NMP-L) is a novel technique that preserves liver grafts under near-physiological conditions while maintaining their normal metabolic activity. This process requires an adequate oxygen supply, typically delivered by packed red blood cells (RBC). We present the first experience using an acellular hemoglobin-based oxygen carrier (HBOC) Hemopure in a human model of NMP-L. Five discarded high-risk human livers were perfused with HBOC-based perfusion fluid and matched to 5 RBC-perfused livers. Perfusion parameters, oxygen extraction, metabolic activity, and histological features were compared during 6 hours of NMP-L. The cytotoxicity of Hemopure was also tested on human hepatic primary cell line cultures using an in vitro model of ischemia reperfusion injury. The vascular flow parameters and the perfusate lactate clearance were similar in both groups. The HBOC-perfused livers extracted more oxygen than those perfused with RBCs (O2 extraction ratio 13.75 vs 9.43 % ×10 per gram of tissue, P = 0.001). In vitro exposure to Hemopure did not alter intracellular levels of reactive oxygen species, and there was no increase in apoptosis or necrosis observed in any of the tested cell lines. Histological findings were comparable between groups. There was no evidence of histological damage caused by Hemopure. Hemopure can be used as an alternative oxygen carrier to packed red cells in NMP-L perfusion fluid.

Defining process design space for monoclonal antibody cell culture.

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Abu-Absi, Susan Fugett; Yang, LiYing; Thompson, Patrick; Jiang, Canping; Kandula, Sunitha; Schilling, Bernhard; Shukla, Abhinav A

2010-08-15

The concept of design space has been taking root as a foundation of in-process control strategies for biopharmaceutical manufacturing processes. During mapping of the process design space, the multidimensional combination of operational variables is studied to quantify the impact on process performance in terms of productivity and product quality. An efficient methodology to map the design space for a monoclonal antibody cell culture process is described. A failure modes and effects analysis (FMEA) was used as the basis for the process characterization exercise. This was followed by an integrated study of the inoculum stage of the process which includes progressive shake flask and seed bioreactor steps. The operating conditions for the seed bioreactor were studied in an integrated fashion with the production bioreactor using a two stage design of experiments (DOE) methodology to enable optimization of operating conditions. A two level Resolution IV design was followed by a central composite design (CCD). These experiments enabled identification of the edge of failure and classification of the operational parameters as non-key, key or critical. In addition, the models generated from the data provide further insight into balancing productivity of the cell culture process with product quality considerations. Finally, process and product-related impurity clearance was evaluated by studies linking the upstream process with downstream purification. Production bioreactor parameters that directly influence antibody charge variants and glycosylation in CHO systems were identified.

Pulmonary artery perfusion versus no pulmonary perfusion during cardiopulmonary bypass in patients with COPD

DEFF Research Database (Denmark)

Buggeskov, Katrine B; Sundskard, Martin M; Jonassen, Thomas

2016-01-01

INTRODUCTION: Absence of pulmonary perfusion during cardiopulmonary bypass (CPB) may be associated with reduced postoperative oxygenation. Effects of active pulmonary artery perfusion were explored in patients with chronic obstructive pulmonary disease (COPD) undergoing cardiac surgery. METHODS: 90...... perfusion with normothermic oxygenated blood during cardiopulmonary bypass appears to improve postoperative oxygenation in patients with COPD undergoing cardiac surgery. Pulmonary artery perfusion with hypothermic HTK solution does not seem to improve postoperative oxygenation. TRIAL REGISTRATION NUMBER...

Define of internal recirculation coefficient for biological wastewater treatment in anoxic and aerobic bioreactors

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Rossinskyi, Volodymyr

2018-02-01

The biological wastewater treatment technologies in anoxic and aerobic bioreactors with recycle of sludge mixture are used for the effective removal of organic compounds from wastewater. The change rate of sludge mixture recirculation between bioreactors leads to a change and redistribution of concentrations of organic compounds in sludge mixture in bioreactors and change hydrodynamic regimes in bioreactors. Determination of the coefficient of internal recirculation of sludge mixture between bioreactors is important for the choice of technological parameters of biological treatment (wastewater treatment duration in anoxic and aerobic bioreactors, flow capacity of recirculation pumps). Determination of the coefficient of internal recirculation of sludge mixture requires integrated consideration of hydrodynamic parameter (flow rate), kinetic parameter (rate of oxidation of organic compounds) and physical-chemical parameter of wastewater (concentration of organic compounds). The conducted numerical experiment from the proposed mathematical equations allowed to obtain analytical dependences of the coefficient of internal recirculation sludge mixture between bioreactors on the concentration of organic compounds in wastewater, the duration of wastewater treatment in bioreactors.

Scale-up from shake flasks to bioreactor, based on power input and Streptomyces lividans morphology, for the production of recombinant APA (45/47 kDa protein) from Mycobacterium tuberculosis.

Science.gov (United States)

Gamboa-Suasnavart, Ramsés A; Marín-Palacio, Luz D; Martínez-Sotelo, José A; Espitia, Clara; Servín-González, Luis; Valdez-Cruz, Norma A; Trujillo-Roldán, Mauricio A

2013-08-01

Culture conditions in shake flasks affect filamentous Streptomyces lividans morphology, as well the productivity and O-mannosylation of recombinant Ala-Pro-rich O-glycoprotein (known as the 45/47 kDa or APA antigen) from Mycobacterium tuberculosis. In order to scale up from previous reported shake flasks to bioreactor, data from the literature on the effect of agitation on morphology of Streptomyces strains were used to obtain gassed volumetric power input values that can be used to obtain a morphology of S. lividans in bioreactor similar to the morphology previously reported in coiled/baffled shake flasks by our group. Morphology of S. lividans was successfully scaled-up, obtaining similar mycelial sizes in both scales with diameters of 0.21 ± 0.09 mm in baffled and coiled shake flasks, and 0.15 ± 0.01 mm in the bioreactor. Moreover, the specific growth rate was successfully scaled up (0.09 ± 0.02 and 0.12 ± 0.01 h(-1), for bioreactors and flasks, respectively), and the recombinant protein productivity measured by densitometry, as well. More interestingly, the quality of the recombinant glycoprotein measured as the amount of mannoses attached to the C-terminal of APA was also scaled- up; with up to five mannose residues in cultures carried out in shake flasks; and six in the bioreactor. However, final biomass concentration was not similar, indicating that although the process can be scaled-up using the power input, others factors like oxygen transfer rate, tip speed or energy dissipation/circulation function can be an influence on bacterial metabolism.

Reverse ventilation--perfusion mismatch

International Nuclear Information System (INIS)

Palmaz, J.C.; Barnett, C.A.; Reich, S.B.; Krumpe, P.E.; Farrer, P.A.

1984-01-01

Patients having lobar airway obstruction or consolidation usually have decreases of both ventilation and perfusion on lung scans. We report three patients in whom hypoxic vasoconstriction was apparently incomplete, resulting in a ''reversed'' ventilation-perfusion mismatch. Perfusion of the hypoxic lobe on the radionuclide scan was associated with metabolic alkalosis, pulmonary venous and pulmonary arterial hypertension in these patients

Long Term Performance of an Arsenite-Oxidizing-Chlorate-Reducing Microbial Consortium in an Upflow Anaerobic Sludge Bed (UASB) Bioreactor

Science.gov (United States)

Sun, Wenjie; Sierra-Alvarez, Reyes; Field, Jim A.

2011-01-01

A chlorate (ClO3−) reducing microbial consortium oxidized arsenite (As(III)) to arsenate (As(V)) in an upflow anaerobic sludge-bed bioreactor over 550 d operation. As(III) was converted with high conversion efficiencies (>98%) at volumetric loadings ranging from 0.45 to 1.92 mmol As/(Lreactor d). The oxidation of As(III) was linked to the complete reduction of ClO3− to Cl− and H2O, as demonstrated by a molar ratio of approximately 3.0 mol As(III) oxidized per mole of Cl− formed and by the greatly lowered ClO3−-reducing capacity without As(III) feeding. An autotrophic enrichment culture was established from the bioreactor biofilm. A 16S rRNA gene clone library indicated that the culture was dominated by Dechloromonas, and Stenotrophomonas as well as genera within the family Comamonadaceae. The results indicate that the oxidation of As(III) to less mobile As(V) utilizing ClO3− as a terminal electron acceptor provides a sustainable bioremediation strategy for arsenic contamination in anaerobic environments. PMID:21333531

Static compression down-regulates N-cadherin expression and facilitates loss of cell phenotype of nucleus pulposus cells in a disc perfusion culture.

Science.gov (United States)

Zhou, Haibo; Shi, Jianmin; Zhang, Chao; Li, Pei

2018-02-28

Mechanical compression often induces degenerative changes of disc nucleus pulposus (NP) tissue. It has been indicated that N-cadherin (N-CDH)-mediated signaling helps to preserve the NP cell phenotype. However, N-CDH expression and the resulting NP-specific phenotype alteration under the static compression and dynamic compression remain unclear. To study the effects of static compression and dynamic compression on N-CDH expression and NP-specific phenotype in an in vitro disc organ culture. Porcine discs were organ cultured in a self-developed mechanically active bioreactor for 7 days and subjected to static or dynamic compression (0.4 MPa for 2 h once per day). The noncompressed discs were used as controls. Compared with the dynamic compression, static compression significantly down-regulated the expression of N-CDH and NP-specific markers (laminin, brachyury, and keratin 19); decreased the Alcian Blue staining intensity, glycosaminoglycan and hydroxyproline contents; and declined the matrix macromolecule (aggrecan and collagen II) expression. Compared with the dynamic compression, static compression causes N-CDH down-regulation, loss of NP-specific phenotype, and the resulting decrease in NP matrix synthesis. © 2018 The Author(s).

Capacity planning for batch and perfusion bioprocesses across multiple biopharmaceutical facilities.

Science.gov (United States)

Siganporia, Cyrus C; Ghosh, Soumitra; Daszkowski, Thomas; Papageorgiou, Lazaros G; Farid, Suzanne S

2014-01-01

Production planning for biopharmaceutical portfolios becomes more complex when products switch between fed-batch and continuous perfusion culture processes. This article describes the development of a discrete-time mixed integer linear programming (MILP) model to optimize capacity plans for multiple biopharmaceutical products, with either batch or perfusion bioprocesses, across multiple facilities to meet quarterly demands. The model comprised specific features to account for products with fed-batch or perfusion culture processes such as sequence-dependent changeover times, continuous culture constraints, and decoupled upstream and downstream operations that permit independent scheduling of each. Strategic inventory levels were accounted for by applying cost penalties when they were not met. A rolling time horizon methodology was utilized in conjunction with the MILP model and was shown to obtain solutions with greater optimality in less computational time than the full-scale model. The model was applied to an industrial case study to illustrate how the framework aids decisions regarding outsourcing capacity to third party manufacturers or building new facilities. The impact of variations on key parameters such as demand or titres on the optimal production plans and costs was captured. The analysis identified the critical ratio of in-house to contract manufacturing organization (CMO) manufacturing costs that led the optimization results to favor building a future facility over using a CMO. The tool predicted that if titres were higher than expected then the optimal solution would allocate more production to in-house facilities, where manufacturing costs were lower. Utilization graphs indicated when capacity expansion should be considered. © 2014 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers.

Disposable bioreactors: maturation into pharmaceutical glycoprotein manufacturing.

Science.gov (United States)

Brecht, René

2009-01-01

Modern biopharmaceutical development is characterised by deep understanding of the structure activity relationship of biological drugs. Therefore, the production process has to be tailored more to the product requirements than to the existing equipment in a certain facility. In addition, the major challenges for the industry are to lower the high production costs of biologics and to shorten the overall development time. The flexibility for providing different modes of operation using disposable bioreactors in the same facility can fulfil these demands and support tailor-made processes.Over the last 10 years, a huge and still increasing number of disposable bioreactors have entered the market. Bioreactor volumes of up to 2,000 L can be handled by using disposable bag systems. Each individual technology has been made available for different purposes up to the GMP compliant production of therapeutic drugs, even for market supply. This chapter summarises disposable technology development over the last decade by comparing the different technologies and showing trends and concepts for the future.

Hydraulic Behavior in The Downflow Hanging Sponge Bioreactor

Directory of Open Access Journals (Sweden)

Izarul Machdar

2016-12-01

Full Text Available Performance efficiency in a Downflow Hanging Sponge (DHS bioreactor is associated with the amount of time that a wastewater remains in the bioreactor. The bioreactor is considered as a plug flow reactor and its hydraulic residence time (HRT depends on the void volume of packing material and the flow rate. In this study, hydraulic behavior of DHS bioreactor was investigated by using tracer method. Two types of sponge module covers, cylindrical plastic frame (module-1 and plastic hair roller (module-2, were investigated and compared. A concentrated NaCl solution used as an inert tracer and input as a pulse at the inlet of DHS bioreactor. Analysis of the residence time distribution (RTD curves provided interpretation of the index distribution or holdup water (active volume, the degree of short-circuiting, number of tanks in series (the plug flow characteristic, and the dispersion number. It was found that the actual HRT was primarily shorter than theoretical HRT of each test. Holdup water of the DHS bioreactor ranged from 60% to 97% and 36% to 60% of module-1 and module-2, respectively. Eventhough module-1 has higher effective volume than module-2, result showed that the dispersion numbers of the two modules were not significant difference. Furthermore, N-values were found larger at a higher flow rate. It was concluded that a DHS bioreactor design should incorporated a combination of water distributor system, higher loading rate at startup process to generate a hydraulic behavior closer to an ideal plug flow.ABSTRAKEfisiensi unjuk kerja bioreactor Downflow Hanging Sponge (DHS berkaitan dengan lamanya waktu tinggal limbah berada di dalam bioreaktor tersebut. Bioreaktor DHS dianggap sebagai seuatu reaktor aliran sumbat (plug flow dimana waktu tinggal hidraulik (HRT tergantung pada volume pori material isian dan laju alir. Dua jenis modul digunakan dalam penelitian ini, yang diberi nama dengan module-1 dan module-2 untuk melihat pengaruh jenis modul

Perfusion dyssynchrony analysis

NARCIS (Netherlands)

Chiribiri, A.; Villa, A.D.M.; Sammut, E.; Breeuwer, M.; Nagel, E.

2015-01-01

AIMS: We sought to describe perfusion dyssynchrony analysis specifically to exploit the high temporal resolution of stress perfusion CMR. This novel approach detects differences in the temporal distribution of the wash-in of contrast agent across the left ventricular wall. METHODS AND RESULTS:

Brain perfusion: computed tomography applications

International Nuclear Information System (INIS)

Miles, K.A.

2004-01-01

Within recent years, the broad introduction of fast multi-detector computed tomography (CT) systems and the availability of commercial software for perfusion analysis have made cerebral perfusion imaging with CT a practical technique for the clinical environment. The technique is widely available at low cost, accurate and easy to perform. Perfusion CT is particularly applicable to those clinical circumstances where patients already undergo CT for other reasons, including stroke, head injury, subarachnoid haemorrhage and radiotherapy planning. Future technical developments in multi-slice CT systems may diminish the current limitations of limited spatial coverage and radiation burden. CT perfusion imaging on combined PET-CT systems offers new opportunities to improve the evaluation of patients with cerebral ischaemia or tumours by demonstrating the relationship between cerebral blood flow and metabolism. Yet CT is often not perceived as a technique for imaging cerebral perfusion. This article reviews the use of CT for imaging cerebral perfusion, highlighting its advantages and disadvantages and draws comparisons between perfusion CT and magnetic resonance imaging. (orig.)

Equipment characterization to mitigate risks during transfers of cell culture manufacturing processes.

Science.gov (United States)

Sieblist, Christian; Jenzsch, Marco; Pohlscheidt, Michael

2016-08-01

The production of monoclonal antibodies by mammalian cell culture in bioreactors up to 25,000 L is state of the art technology in the biotech industry. During the lifecycle of a product, several scale up activities and technology transfers are typically executed to enable the supply chain strategy of a global pharmaceutical company. Given the sensitivity of mammalian cells to physicochemical culture conditions, process and equipment knowledge are critical to avoid impacts on timelines, product quantity and quality. Especially, the fluid dynamics of large scale bioreactors versus small scale models need to be described, and similarity demonstrated, in light of the Quality by Design approach promoted by the FDA. This approach comprises an associated design space which is established during process characterization and validation in bench scale bioreactors. Therefore the establishment of predictive models and simulation tools for major operating conditions of stirred vessels (mixing, mass transfer, and shear force.), based on fundamental engineering principles, have experienced a renaissance in the recent years. This work illustrates the systematic characterization of a large variety of bioreactor designs deployed in a global manufacturing network ranging from small bench scale equipment to large scale production equipment (25,000 L). Several traditional methods to determine power input, mixing, mass transfer and shear force have been used to create a data base and identify differences for various impeller types and configurations in operating ranges typically applied in cell culture processes at manufacturing scale. In addition, extrapolation of different empirical models, e.g. Cooke et al. (Paper presented at the proceedings of the 2nd international conference of bioreactor fluid dynamics, Cranfield, UK, 1988), have been assessed for their validity in these operational ranges. Results for selected designs are shown and serve as examples of structured

Biological reduction of nitrate wastewater using fluidized-bed bioreactors

International Nuclear Information System (INIS)

Walker, J.F. Jr.; Hancher, C.W.; Patton, B.D.; Kowalchuk, M.

1981-01-01

There are a number of nitrate-containing wastewater sources, as concentrated as 30 wt % NO 3 - and as large as 2000 m 3 /d, in the nuclear fuel cycle as well as in many commercial processes such as fertilizer production, paper manufacturing, and metal finishing. These nitrate-containing wastewater sources can be successfully biologically denitrified to meet discharge standards in the range of 10 to 20 gN(NO 3 - )/m 3 by the use of a fluidized-bed bioreactor. The major strain of denitrification bacteria is Pseudomonas which was derived from garden soil. In the fluidized-bed bioreactor the bacteria are allowed to attach to 0.25 to 0.50-mm-diam coal particles, which are fluidized by the upward flow of influent wastewater. Maintaining the bacteria-to-coal weight ratio at approximately 1:10 results in a bioreactor bacteria loading of greater than 20,000 g/m 3 . A description is given of the results of two biodenitrification R and D pilot plant programs based on the use of fluidized bioreactors capable of operating at nitrate levels up to 7000 g/m 3 and achieving denitrification rates as high as 80 gN(NO 3 - )/d per liter of empty bioreactor volume. The first of these pilot plant programs consisted of two 0.2-m-diam bioreactors, each with a height of 6.3 m and a volume of 208 liters, operating in series. The second pilot plant was used to determine the diameter dependence of the reactors by using a 0.5-m-diam reactor with a height of 6.3 m and a volume of 1200 liters. These pilot plants operated for a period of six months and two months respectively, while using both a synthetic waste and the actual waste from a gaseous diffusion plant operated by Goodyear Atomic Corporation

Development of a new bioprocess scheme using frozen seed train intermediates to initiate CHO cell culture manufacturing campaigns.

Science.gov (United States)

Seth, Gargi; Hamilton, Robert W; Stapp, Thomas R; Zheng, Lisa; Meier, Angela; Petty, Krista; Leung, Stephenie; Chary, Srikanth

2013-05-01

Agility to schedule and execute cell culture manufacturing campaigns quickly in a multi-product facility will play a key role in meeting the growing demand for therapeutic proteins. In an effort to shorten campaign timelines, maximize plant flexibility and resource utilization, we investigated the initiation of cell culture manufacturing campaigns using CHO cells cryopreserved in large volume bags in place of the seed train process flows that are conventionally used in cell culture manufacturing. This approach, termed FASTEC (Frozen Accelerated Seed Train for Execution of a Campaign), involves cultivating cells to high density in a perfusion bioreactor, and cryopreserving cells in multiple disposable bags. Each run for a manufacturing campaign would then come from a thaw of one or more of these cryopreserved bags. This article reviews the development and optimization of individual steps of the FASTEC bioprocess scheme: scaling up cells to greater than 70 × 10(6) cells/mL and freezing in bags with an optimized controlled rate freezing protocol and using a customized rack configuration. Flow cytometry analysis was also employed to understand the recovery of CHO cells following cryopreservation. Extensive development data were gathered to ensure that the quantity and quality of the drug manufactured using the FASTEC bioprocess scheme was acceptable compared to the conventional seed train process flow. The result of offering comparable manufacturing options offers flexibility to the cell culture manufacturing network. Copyright © 2012 Wiley Periodicals, Inc.

Production of the biopesticide azadirachtin by hairy root cultivation of Azadirachta indica in liquid-phase bioreactors.

Science.gov (United States)

Srivastava, Smita; Srivastava, Ashok K

2013-11-01

Batch cultivation of Azadirachta indica hairy roots was carried out in different liquid-phase bioreactor configurations (stirred-tank, bubble column, bubble column with polypropylene basket, and polyurethane foam disc as root supports) to investigate possible scale-up of the A. indica hairy root culture for in vitro production of the biopesticide azadirachtin. The hairy roots failed to grow in the conventional bioreactor designs (stirred tank and bubble column). However, modified bubble column reactor (with polyurethane foam as root support) configuration facilitated high-density culture of A. indica hairy roots with a biomass production of 9.2 g l(-1)dry weight and azadirachtin yield of 3.2 mg g(-1) leading to a volumetric productivity of azadirachtin as 1.14 mg l(-1) day(-1). The antifeedant activity in the hairy roots was also evaluated by no choice feeding tests with known concentrations of the hairy root powder and its solvent extract separately on the desert locust Schistocerca gregaria. The hairy root powder and its solvent extract demonstrated a high level of antifeedant activity (with an antifeedant index of 97 % at a concentration of 2 % w/v and 83 % at a concentration of 0.05 % (w/v), respectively, in ethanol).

Dynamic cell culture system (7-IML-1)

Science.gov (United States)

Cogoli, Augusto

1992-01-01

This experiment is one of the Biorack experiments being flown on the International Microgravity Laboratory 1 (MIL-1) mission as part of an investigation studying cell proliferation and performance in space. One of the objectives of this investigation is to assess the potential benefits of bioprocessing in space with the ultimate goal of developing a bioreactor for continuous cell cultures in space. This experiment will test the operation of an automated culture chamber that was designed for use in a Bioreactor in space. The device to be tested is called the Dynamic Cell Culture System (DCCS). It is a simple device in which media are renewed or chemicals are injected automatically, by means of osmotic pumps. This experiment uses four Type I/O experiment containers. One DCCS unit, which contains a culture chamber with renewal of medium and a second chamber without a medium supply fits in each container. Two DCCS units are maintained under zero gravity conditions during the on-orbit period. The other two units are maintained under 1 gh conditions in a 1 g centrifuge. The schedule for incubator transfer is given.

Use of disposable reactors to generate inoculum cultures for E. coli production fermentations.

Science.gov (United States)

Mahajan, Ekta; Matthews, Timothy; Hamilton, Ryan; Laird, Michael W

2010-01-01

Disposable technology is being used more each year in the biotechnology industry. Disposable bioreactors allow one to avoid expenses associated with cleaning, assembly and operations, as well as equipment validation. The WAVE bioreactor is well established for Chinese Hamster Ovary (CHO) production, however, it has not yet been thoroughly tested for E. coli production because of the high oxygen demand and temperature maintenance requirements of that platform. The objective of this study is to establish a robust process to generate inoculum for E. coli production fermentations in a WAVE bioreactor. We opted not to evaluate the WAVE system for production cultures because of the high cell densities required in our current E. coli production processes. Instead, the WAVE bioreactor 20/50 system was evaluated at laboratory scale (10-L) to generate inoculum with target optical densities (OD(550)) of 15 within 7-9 h (pre-established target for stainless steel fermentors). The maximum settings for rock rate (40 rpm) and angle (10.5) were used to maximize mass transfer. The gas feed was also supplemented with additional oxygen to meet the high respiratory demand of the culture. The results showed that the growth profiles for the inoculum cultures were similar to those obtained from conventional stainless steel fermentors. These inoculum cultures were subsequently inoculated into 10-L working volume stainless steel fermentors to evaluate the inocula performance of two different production systems during recombinant protein production. The results of these production cultures using WAVE inocula showed that the growth and recombinant protein production was comparable to the control data set. Furthermore, an economic analysis showed that the WAVE system would require less capital investment for installation and operating expenses would be less than traditional stainless steel systems. (c) 2010 American Institute of Chemical Engineers

A capillary-based perfusion phantom for simulation of brain perfusion for MRI

International Nuclear Information System (INIS)

Maciak, A.; Kronfeld, A.; Mueller-Forell, W.; Wille, C.; Kempski, O.; Stoeter, P.

2010-01-01

Purpose: The measurement of the CBF is a non-standardized procedure and there are no reliable gold standards. This abstract shows a capillary-based perfusion-phantom for CE-DSC-MRI. It has equivalent flow properties to those within the tissue capillary system of the human brain and allows the validation of the Siemens Perfusion (MR) software. Materials and Methods: The perfusion phantom consists of a dialyzer for the simulation of the capillary system, a feeding tube for simulation of the AIF and a pulsatile pump for simulation of the heart. Using this perfusion phantom, the exact determination of the gold standard CBF due to the well-known geometry of the phantom is easy. It was validated based on different perfusion measurements. These measurements were investigated with standard software (Siemens Perfusion MR). The software determined the CBF within the capillary system. Based on this CBF, a comparison to the gold standard was made with several different flow speeds. After AIF selection, a total of 726 CBF data points were automatically extracted by the software. Results: This results in a comparison of the gold standard CBF to these 726 CBF values. Therefore, a reproducible and reliable deviation estimation between gold standard CBF and measured CBF using the software was computed. It can be shown that the deviation between gold standard and software-based evaluation ranges between 1 and 31 %. Conclusion: There is no significance for any correlation between flow speed and amount of deviation. The mean measured CBF is 11.4 % higher than the gold standard CBF (p-value < 0.001). Using this kind of perfusion-phantom, the validation of different software systems allows reliable conclusions about their quality. (orig.)

Perfusion CT in acute stroke

International Nuclear Information System (INIS)

Eckert, Bernd; Roether, Joachim; Fiehler, Jens; Thomalla, Goetz

2015-01-01

Modern multislice CT scanners enable multimodal protocols including non-enhanced CT, CT angiography, and CT perfusion. A 64-slice CT scanner provides 4-cm coverage. To cover the whole brain, a 128 - 256-slice scanner is needed. The use of perfusion CT requires an optimized scan protocol in order to reduce exposure to radiation. As compared to non-enhanced CT and CT angiography, the use of CT perfusion increases detection rates of cerebral ischemia, especially small cortical ischemic lesions, while the detection of lacunar and infratentorial stroke lesions remains limited. Perfusion CT enables estimation of collateral flow in acute occlusion of large intra- or extracranial arteries. Currently, no established reliable thresholds are available for determining infarct core and penumbral tissue by CT perfusion. Moreover, perfusion parameters depend on the processing algorithms and the software used for calculation. However, a number of studies point towards a reduction of cerebral blood volume (CBV) below 2 ml/100 g as a critical threshold that identifies infarct core. Large CBV lesions are associated with poor outcome even in the context of recanalization. The extent of early ischemic signs on non-enhanced CT remains the main parameter from CT imaging to guide acute reperfusion treatment. Nevertheless, perfusion CT increases diagnostic and therapeutic certainty in the acute setting. Similar to stroke MRI, perfusion CT enables the identification of tissue at risk of infarction by the mismatch between infarct core and the larger area of critical hypoperfusion. Further insights into the validity of perfusion parameters are expected from ongoing trials of mechanical thrombectomy in stroke.

Volume perfusion CT imaging of cerebral vasospasm: diagnostic performance of different perfusion maps

Energy Technology Data Exchange (ETDEWEB)

Othman, Ahmed E. [RWTH Aachen University, Department of Diagnostic and Interventional Neuroradiology, Aachen (Germany); Eberhard Karls University Tuebingen, University Hospital Tuebingen, Department for Diagnostic and Interventional Radiology, Tuebingen (Germany); Afat, Saif; Nikoubashman, Omid; Mueller, Marguerite; Wiesmann, Martin; Brockmann, Carolin [RWTH Aachen University, Department of Diagnostic and Interventional Neuroradiology, Aachen (Germany); Schubert, Gerrit Alexander [RWTH Aachen University, Department of Neurosurgery, Aachen (Germany); Bier, Georg [Eberhard Karls University Tuebingen, University Hospital Tuebingen, Department for Diagnostic and Interventional Neuroradiology, Tuebingen (Germany); Brockmann, Marc A. [RWTH Aachen University, Department of Diagnostic and Interventional Neuroradiology, Aachen (Germany); University Hospital Mainz, Department of Neuroradiology, Mainz (Germany)

2016-08-15

In this study, we aimed to evaluate the diagnostic performance of different volume perfusion CT (VPCT) maps regarding the detection of cerebral vasospasm compared to angiographic findings. Forty-one datasets of 26 patients (57.5 ± 10.8 years, 18 F) with subarachnoid hemorrhage and suspected cerebral vasospasm, who underwent VPCT and angiography within 6 h, were included. Two neuroradiologists independently evaluated the presence and severity of vasospasm on perfusion maps on a 3-point Likert scale (0 - no vasospasm, 1 - vasospasm affecting <50 %, 2 - vasospasm affecting >50 % of vascular territory). A third neuroradiologist independently assessed angiography for the presence and severity of vasospasm on a 3-point Likert scale (0 - no vasospasm, 1 - vasospasm affecting < 50 %, 2 - vasospasm affecting > 50 % of vessel diameter). Perfusion maps of cerebral blood volume (CBV), cerebral blood flow (CBF), mean transit time (MTT), and time to drain (TTD) were evaluated regarding diagnostic accuracy for cerebral vasospasm with angiography as reference standard. Correlation analysis of vasospasm severity on perfusion maps and angiographic images was performed. Furthermore, inter-reader agreement was assessed regarding findings on perfusion maps. Diagnostic accuracy for TTD and MTT was significantly higher than for all other perfusion maps (TTD, AUC = 0.832; MTT, AUC = 0.791; p < 0.001). TTD revealed higher sensitivity than MTT (p = 0.007). The severity of vasospasm on TTD maps showed significantly higher correlation levels with angiography than all other perfusion maps (p ≤ 0.048). Inter-reader agreement was (almost) perfect for all perfusion maps (kappa ≥ 0.927). The results of this study indicate that TTD maps have the highest sensitivity for the detection of cerebral vasospasm and highest correlation with angiography regarding the severity of vasospasm. (orig.)

Process for whole cell saccharification of lignocelluloses to sugars using a dual bioreactor system

Science.gov (United States)

Lu, Jue [Okemos, MI; Okeke, Benedict [Montgomery, AL

2012-03-27

The present invention describes a process for saccharification of lignocelluloses to sugars using whole microbial cells, which are enriched from cultures inoculated with paper mill waste water, wood processing waste and soil. A three-member bacterial consortium is selected as a potent microbial inocula and immobilized on inedible plant fibers for biomass saccharification. The present invention further relates the design of a dual bioreactor system, with various biocarriers for enzyme immobilization and repeated use. Sugars are continuously removed eliminating end-product inhibition and consumption by cell.

In vitro azadirachtin production by hairy root cultivation of Azadirachta indica in nutrient mist bioreactor.

Science.gov (United States)

Srivastava, Smita; Srivastava, A K

2012-01-01

Azadirachtin, a well-known biopesticide is a secondary metabolite conventionally extracted from the seeds of Azadirachta indica. The present study involved in vitro azadirachtin production by developing hairy roots of A. indica via Agrobacterium rhizogenes-mediated transformation of A. indica explants. Liquid culture of hairy roots was established in shake flask to study the kinetics of growth and azadirachtin production. A biomass production of 13.3 g/L dry weight (specific growth rate of 0.7 day(-1)) was obtained after 25 days of cultivation period with an azadirachtin yield of 3.3 mg/g root biomass. To overcome the mass transfer limitation in conventionally used liquid-phase reactors, batch cultivation of hairy roots was carried out in gas-phase reactors (nutrient spray and nutrient mist bioreactor) to investigate the possible scale-up of A. indica hairy root culture. The nano-size nutrient mist particles generated from the nozzle of the nutrient mist bioreactor could penetrate till the inner core of the inoculated root matrix, facilitating uniform growth during high-density cultivation of hairy roots. A biomass production of 9.8 g/L dry weight with azadirachtin accumulation of 2.8 mg/g biomass (27.4 mg/L) could be achieved in 25 days of batch cultivation period, which was equivalent to a volumetric productivity of 1.09 mg/L per day of azadirachtin.

Radiation response of perfused tracheal sections

International Nuclear Information System (INIS)

Ford, J.R.; Maslowski, A.J.; Braby, L.A.

2003-01-01

Full text: A model of respiratory tissue using a perfusion culture system is being developed. We are using this system to quantify the effects of normal tissue architecture and the interaction of epithelial cells with other cell types on radiation-induced bystander effects. Tracheal tissue taken from young adult male Fischer 344 rats is imbedded in a growth factor enriched agarose matrix. The chamber is designed to allow growth medium to periodically wash the epithelial surface of the tracheal lumen while maintaining the air-interface that is necessary for the normal differentiation of the epithelium. In preliminary experiments with rat trachea we have shown that a differentiated epithelial lining can be maintained for several days. Cells can be obtained for a number of different cell culture assays for endpoints such as survival and preneoplastic transformation after irradiation

Standardized perfusion value of the esophageal carcinoma and its correlation with quantitative CT perfusion parameter values

Energy Technology Data Exchange (ETDEWEB)

Djuric-Stefanovic, A., E-mail: avstefan@eunet.rs [Faculty of Medicine, University of Belgrade, Belgrade (Serbia); Unit of Digestive Radiology (First University Surgical Clinic), Center of Radiology and MR, Clinical Center of Serbia, Belgrade (Serbia); Saranovic, Dj., E-mail: crvzve4@gmail.com [Faculty of Medicine, University of Belgrade, Belgrade (Serbia); Unit of Digestive Radiology (First University Surgical Clinic), Center of Radiology and MR, Clinical Center of Serbia, Belgrade (Serbia); Sobic-Saranovic, D., E-mail: dsobic2@gmail.com [Faculty of Medicine, University of Belgrade, Belgrade (Serbia); Center of Nuclear Medicine, Clinical Center of Serbia, Belgrade (Serbia); Masulovic, D., E-mail: draganmasulovic@yahoo.com [Faculty of Medicine, University of Belgrade, Belgrade (Serbia); Unit of Digestive Radiology (First University Surgical Clinic), Center of Radiology and MR, Clinical Center of Serbia, Belgrade (Serbia); Artiko, V., E-mail: veraart@beotel.rs [Faculty of Medicine, University of Belgrade, Belgrade (Serbia); Center of Nuclear Medicine, Clinical Center of Serbia, Belgrade (Serbia)

2015-03-15

Purpose: Standardized perfusion value (SPV) is a universal indicator of tissue perfusion, normalized to the whole-body perfusion, which was proposed to simplify, unify and allow the interchangeability among the perfusion measurements and comparison between the tumor perfusion and metabolism. The aims of our study were to assess the standardized perfusion value (SPV) of the esophageal carcinoma, and its correlation with quantitative CT perfusion measurements: blood flow (BF), blood volume (BV), mean transit time (MTT) and permeability surface area product (PS) of the same tumor volume samples, which were obtained by deconvolution-based CT perfusion analysis. Methods: Forty CT perfusion studies of the esophageal cancer were analyzed, using the commercial deconvolution-based CT perfusion software (Perfusion 3.0, GE Healthcare). The SPV of the esophageal tumor and neighboring skeletal muscle were correlated with the corresponding mean tumor and muscle quantitative CT perfusion parameter values, using Spearman's rank correlation coefficient (r{sub S}). Results: Median SPV of the esophageal carcinoma (7.1; range: 2.8–13.4) significantly differed from the SPV of the skeletal muscle (median: 1.0; range: 0.4–2.4), (Z = −5.511, p < 0.001). The cut-off value of the SPV of 2.5 enabled discrimination of esophageal cancer from the skeletal muscle with sensitivity and specificity of 100%. SPV of the esophageal carcinoma significantly correlated with corresponding tumor BF (r{sub S} = 0.484, p = 0.002), BV (r{sub S} = 0.637, p < 0.001) and PS (r{sub S} = 0.432, p = 0.005), and SPV of the skeletal muscle significantly correlated with corresponding muscle BF (r{sub S} = 0.573, p < 0.001), BV (r{sub S} = 0.849, p < 0.001) and PS (r{sub S} = 0.761, p < 0.001). Conclusions: We presented a database of the SPV for the esophageal cancer and proved that SPV of the esophageal neoplasm significantly differs from the SPV of the skeletal muscle, which represented a sample of healthy

Capacity Planning for Batch and Perfusion Bioprocesses Across Multiple Biopharmaceutical Facilities

Science.gov (United States)

Siganporia, Cyrus C; Ghosh, Soumitra; Daszkowski, Thomas; Papageorgiou, Lazaros G; Farid, Suzanne S

2014-01-01

Production planning for biopharmaceutical portfolios becomes more complex when products switch between fed-batch and continuous perfusion culture processes. This article describes the development of a discrete-time mixed integer linear programming (MILP) model to optimize capacity plans for multiple biopharmaceutical products, with either batch or perfusion bioprocesses, across multiple facilities to meet quarterly demands. The model comprised specific features to account for products with fed-batch or perfusion culture processes such as sequence-dependent changeover times, continuous culture constraints, and decoupled upstream and downstream operations that permit independent scheduling of each. Strategic inventory levels were accounted for by applying cost penalties when they were not met. A rolling time horizon methodology was utilized in conjunction with the MILP model and was shown to obtain solutions with greater optimality in less computational time than the full-scale model. The model was applied to an industrial case study to illustrate how the framework aids decisions regarding outsourcing capacity to third party manufacturers or building new facilities. The impact of variations on key parameters such as demand or titres on the optimal production plans and costs was captured. The analysis identified the critical ratio of in-house to contract manufacturing organization (CMO) manufacturing costs that led the optimization results to favor building a future facility over using a CMO. The tool predicted that if titres were higher than expected then the optimal solution would allocate more production to in-house facilities, where manufacturing costs were lower. Utilization graphs indicated when capacity expansion should be considered. © 2013 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 30:594–606, 2014 PMID:24376262

Nuclear magnetic resonance of perfused tissue

International Nuclear Information System (INIS)

Harpen, M.D.; Allison, R.C.

1986-01-01

The effect of perfusion on the NMR signal observed in NMR imaging is studied in a phantom and in two isolated perfused canine lungs. It is observed that perfusion in tissue has little effect on longitudinal relaxation times. Transverse relaxation rates are observed to correlate linearly with rates of perfusion, in accordance with a model presented. (author)

Dynamic CT myocardial perfusion imaging identifies early perfusion abnormalities in diabetes and hypertension : Insights from a multicenter registry

NARCIS (Netherlands)

Vliegenthart, Rozemarijn; De Cecco, Carlo N.; Wichmann, Julian L.; Meinel, Felix G.; Pelgrim, Gert Jan; Tesche, Christian; Ebersberger, Ullrich; Pugliese, Francesca; Bamberg, Fabian; Choe, Yeon Hyeon; Wang, Yining; Schoepf, U. Joseph

2016-01-01

Background: To identify patients with early signs of myocardial perfusion reduction, a reference base for perfusion measures is needed. Objective: To analyze perfusion parameters derived from dynamic computed tomography perfusion imaging (CTPI) in patients with suspected coronary artery disease

Multi-membrane chitosan hydrogels as chondrocytic cell bioreactors.

Science.gov (United States)

Ladet, S G; Tahiri, K; Montembault, A S; Domard, A J; Corvol, M-T M

2011-08-01

We investigated the bioactivity of new chitosan-based multi-membrane hydrogel (MMH) architectures towards chondrocyte-like cells. The microstructure of the hydrogels constituting the membranes precludes any living cell penetration, whereas their lower scale architecture allows the protein diffusion. The biological behavior of chondrocytes implanted within the MMH inter-membrane spaces was studied for 45 days in culture. Chondrocytes formed cell aggregates and proliferated without loosing their chondrogenic phenotype as illustrated by collagen II and aggrecan expressions at the mRNA and protein levels. Cells produced neo-formed alcyan blue matrix proteins filling MMH interspaces. The HiF-2α/SOX9 pattern of expression suggested that the elevated chondrocytic phenotype in MMH could be related to a better hypoxic local environment than in classical culture conditions. Pro-inflammatory markers were not expressed during the period of culture. The low level of nitric oxide accumulation within the inter-membrane spaces and in the incubation medium implied that chitosan consumed nitrites produced by entrapped chondrocytes, in relation with the decrease of its molecular weight of 50%. Our data suggest that MMH structures may be considered as complex chondrocytic cell bioreactors; "active decoys of biological media", potentially promising for various biomedical applications like the inter-vertebral disk replacement. Copyright © 2011 Elsevier Ltd. All rights reserved.

Review of nonconventional bioreactor technology

Energy Technology Data Exchange (ETDEWEB)

Turick, C.E.; Mcllwain, M.E.

1993-09-01

Biotechnology will significantly affect many industrial sectors in the future. Industrial sectors that will be affected include pharmaceutical, chemical, fuel, agricultural, and environmental remediation. Future research is needed to improve bioprocessing efficiency and cost-effectiveness in order to compete with traditional technologies. This report describes recent advances in bioprocess technologies and bioreactor designs and relates them to problems encountered in many industrial bioprocessing operations. The primary focus is directed towards increasing gas and vapor transfer for enhanced bioprocess kinetics as well as unproved by-product separation and removal. The advantages and disadvantages of various conceptual designs such as hollow-fiber, gas-phase, hyperbaric/hypobaric, and electrochemical bioreactors are also discussed. Specific applications that are intended for improved bioprocesses include coal desulfurization, coal liquefaction, soil bioremediation, biomass conversion to marketable chemicals, biomining, and biohydrometallurgy as well as bioprocessing of gases and vapors.

The Role of Bioreactors in Ligament and Tendon Tissue Engineering.

Science.gov (United States)

Mace, James; Wheelton, Andy; Khan, Wasim S; Anand, Sanj

2016-01-01

Bioreactors are pivotal to the emerging field of tissue engineering. The formation of neotissue from pluripotent cell lineages potentially offers a source of tissue for clinical use without the significant donor site morbidity associated with many contemporary surgical reconstructive procedures. Modern bioreactor design is becoming increasingly complex to provide a both an expandable source of readily available pluripotent cells and to facilitate their controlled differentiation into a clinically applicable ligament or tendon like neotissue. This review presents the need for such a method, challenges in the processes to engineer neotissue and the current designs and results of modern bioreactors in the pursuit of engineered tendon and ligament.

Purification and characterization of two DyP isozymes from Thanatephorus cucumeris Dec 1 specifically expressed in an air-membrane surface bioreactor.

Science.gov (United States)

Shimokawa, Takuya; Shoda, Makoto; Sugano, Yasushi

2009-02-01

DyP isozymes (DyP2 and DyP3) from the culture fluid of the fungus Thanatephorus cucumeris Dec 1 by air-membrane surface bioreactor were purified and characterized. The characteristics of DyP2 were almost the same as those of a recombinant DyP reported previously, but different from DyP3.

Methodology for ventilation/perfusion SPECT

DEFF Research Database (Denmark)

Bajc, Marika; Neilly, Brian; Miniati, Massimo

2010-01-01

radiolabeled liquid aerosols are not restricted to the presence of obstructive lung disease. Radiolabeled macroaggregated human albumin is the imaging agent of choice for perfusion scintigraphy. An optimal combination of nuclide activities and acquisition times for ventilation and perfusion, collimators......Ventilation/perfusion single-photon emission computed tomography (V/Q SPECT) is the scintigraphic technique of choice for the diagnosis of pulmonary embolism and many other disorders that affect lung function. Data from recent ventilation studies show that the theoretic advantages of Technegas over......, and imaging matrix yields an adequate V/Q SPECT study in approximately 20 minutes of imaging time. The recommended protocol based on the patient remaining in an unchanged position during the initial ventilation study and the perfusion study allows presentation of matching ventilation and perfusion slices...

Treating domestic sewage by Integrated Inclined-Plate-Membrane bio-reactor

Science.gov (United States)

Song, Li Ming; Wang, Zi; Chen, Lei; Zhong, Min; Dong, Zhan Feng

2017-12-01

Membrane fouling shorten the service life of the membrane and increases aeration rate for membrane surface cleaning. Two membrane bio-reactors, one for working and another for comparing, were set up to evaluate the feasibility of alleviating membrane fouling and improving wastewater treatment efficiency by integrating inclined-plate precipitation and membrane separation. The result show that: (1) Inclined-plate in reactor had a good effect on pollutant removal of membrane bioreactor. The main role of inclined-plate is dividing reactor space and accelerating precipitation. (2) Working reactor have better performance in COD, TN and TP removal, which can attribute to that working reactor (integrated inclined-plate-Membrane bioreactor) takes both advantages of membrane separation and biological treatment. When influent COD, TP and TN concentration is 163-248 mg/L, 2.08-2.81 mg/L and 24.38-30.49 mg/L in working reactor, effluent concentration is 27-35 mg/L, 0.53-0.59 mg/L and 11.28-11.56 mg/L, respectively. (3) Membrane fouling was well alleviated in integrated inclined-plate-Membrane bioreactor, and membrane normal service time is significantly longer than that in comparing reactor, which can attribute to accelerating precipitation of inclined-plate. In summary, integrated inclined-plate-Membrane bioreactor is a promising technology to alleviating membrane fouling and improving wastewater treatment efficiency, having good performance and bright future in application.

Hepatic differentiation of human iPSCs in different 3D models: A comparative study.

Science.gov (United States)

Meier, Florian; Freyer, Nora; Brzeszczynska, Joanna; Knöspel, Fanny; Armstrong, Lyle; Lako, Majlinda; Greuel, Selina; Damm, Georg; Ludwig-Schwellinger, Eva; Deschl, Ulrich; Ross, James A; Beilmann, Mario; Zeilinger, Katrin

2017-12-01

Human induced pluripotent stem cells (hiPSCs) are a promising source from which to derive distinct somatic cell types for in vitro or clinical use. Existent protocols for hepatic differentiation of hiPSCs are primarily based on 2D cultivation of the cells. In the present study, the authors investigated the generation of hiPSC-derived hepatocyte-like cells using two different 3D culture systems: A 3D scaffold-free microspheroid culture system and a 3D hollow-fiber perfusion bioreactor. The differentiation outcome in these 3D systems was compared with that in conventional 2D cultures, using primary human hepatocytes as a control. The evaluation was made based on specific mRNA expression, protein secretion, antigen expression and metabolic activity. The expression of α-fetoprotein was lower, while cytochrome P450 1A2 or 3A4 activities were higher in the 3D culture systems as compared with the 2D differentiation system. Cells differentiated in the 3D bioreactor showed an increased expression of albumin and hepatocyte nuclear factor 4α, as well as secretion of α-1-antitrypsin as compared with the 2D differentiation system, suggesting a higher degree of maturation. In contrast, the 3D scaffold-free microspheroid culture provides an easy and robust method to generate spheroids of a defined size for screening applications, while the bioreactor culture model provides an instrument for complex investigations under physiological-like conditions. In conclusion, the present study introduces two 3D culture systems for stem cell derived hepatic differentiation each demonstrating advantages for individual applications as well as benefits in comparison with 2D cultures.

Hydrostatic determinants of cerebral perfusion

International Nuclear Information System (INIS)

Wagner, E.M.; Traystman, R.J.

1986-01-01

We examined the cerebral blood flow response to alterations in perfusion pressure mediated through decreases in mean arterial pressure, increases in cerebrospinal fluid (CSF) pressure, and increases in jugular venous (JV) pressure in 42 pentobarbital anesthetized dogs. Each of these three pressures was independently controlled. Cerebral perfusion pressure was defined as mean arterial pressure minus JV or CSF pressure, depending on which was greater. Mean hemispheric blood flow was measured with the radiolabeled microsphere technique. Despite 30-mm Hg reductions in mean arterial pressure or increases in CSF or JV pressure, CBF did not change as long as the perfusion pressure remained greater than approximately 60 mm Hg. However, whenever perfusion pressure was reduced to an average of 48 mm Hg, cerebral blood flow decreased 27% to 33%. These results demonstrate the capacity of the cerebral vascular bed to respond similarly to changes in the perfusion pressure gradient obtained by decreasing mean arterial pressure, increasing JV pressure or increasing CSF pressure, and thereby support the above definition of cerebral perfusion pressure

Culture of equine fibroblast-like synoviocytes on synthetic tissue scaffolds towards meniscal tissue engineering: a preliminary cell-seeding study

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Jennifer J. Warnock

2014-04-01

Full Text Available Introduction. Tissue engineering is a new methodology for addressing meniscal injury or loss. Synovium may be an ideal source of cells for in vitro meniscal fibrocartilage formation, however, favorable in vitro culture conditions for synovium must be established in order to achieve this goal. The objective of this study was to determine cellularity, cell distribution, and extracellular matrix (ECM formation of equine fibroblast-like synoviocytes (FLS cultured on synthetic scaffolds, for potential application in synovium-based meniscal tissue engineering. Scaffolds included open-cell poly-L-lactic acid (OPLA sponges and polyglycolic acid (PGA scaffolds cultured in static and dynamic culture conditions, and PGA scaffolds coated in poly-L-lactic (PLLA in dynamic culture conditions.Materials and Methods. Equine FLS were seeded on OPLA and PGA scaffolds, and cultured in a static environment or in a rotating bioreactor for 12 days. Equine FLS were also seeded on PGA scaffolds coated in 2% or 4% PLLA and cultured in a rotating bioreactor for 14 and 21 days. Three scaffolds from each group were fixed, sectioned and stained with Masson’s Trichrome, Safranin-O, and Hematoxylin and Eosin, and cell numbers and distribution were analyzed using computer image analysis. Three PGA and OPLA scaffolds from each culture condition were also analyzed for extracellular matrix (ECM production via dimethylmethylene blue (sulfated glycosaminoglycan assay and hydroxyproline (collagen assay. PLLA coated PGA scaffolds were analyzed using double stranded DNA quantification as areflection of cellularity and confocal laser microscopy in a fluorescent cell viability assay.Results. The highest cellularity occurred in PGA constructs cultured in a rotating bioreactor, which also had a mean sulfated glycosaminoglycan content of 22.3 µg per scaffold. PGA constructs cultured in static conditions had the lowest cellularity. Cells had difficulty adhering to OPLA and the PLLA

Filtration characteristics in membrane bioreactors

NARCIS (Netherlands)

Evenblij, H.

2006-01-01

Causes of and remedies for membrane fouling in Membrane Bioreactors for wastewater treatment are only poorly understood and described in scientific literature. A Filtration Characterisation Installation and a measurement protocol were developed with the aim of a) unequivocally determination and

Forecasting the settlement of a bioreactor landfill based on gas pressure changes.

Science.gov (United States)

Qiu, Gang; Li, Liang; Sun, Hongjun

2013-10-01

In order to study the influence of settlement under gas pressure in bioreactor landfill, the landfill is simplified as a one-way gas seepage field, combining Darcy's Law, the gas equation of state, and the principle of effective stress and fluid dynamics of porous media theory. First assume that the bioreactor landfill leachate is fully recharged on the basis of gas mass conservation, then according to the changes in gas pressure (inside the landfill and surrounding atmosphere) during the gas leakage time and settlement in the landfill, establish a numerical model of bioreactor landfill settlement under the action of the gas pressure, and use the finite difference method to solve it. Through a case study, the model's improved prediction of the settlement of bioreactor landfill is demonstrated.

Stoichiometric and kinetic analysis of extreme halophilic Archaea on various substrates in a corrosion resistant bioreactor.

Science.gov (United States)

Lorantfy, Bettina; Seyer, Bernhard; Herwig, Christoph

2014-01-25

Extreme halophilic Archaea are extremophile species which can thrive in hypersaline environments of up to 3-5 M sodium chloride concentration. Although their ecology and physiology are widely identified on the microbiological level, little emphasis has been laid on quantitative bioprocess development with extreme halophiles. The goal of this study was to establish, on the one hand, a methodological basis for quantitative bioprocess analysis of extreme halophilic Archaea with an extreme halophilic strain as an example. Firstly, as a novel usage, a corrosion resistant bioreactor setup for extreme halophiles has been implemented. Then, paying special attention to total bioprocess quantification approaches, an indirect method for biomass quantification using on-line process signals was introduced. Subsequently, robust quantitative data evaluation methods for halophiles could be developed, providing defined and controlled cultivation conditions in the bioreactor and therefore obtaining suitable quality of on-line as well as off-line datasets. On the other hand, new physiological results of extreme halophiles in bioreactor have also been obtained based on the quantitative methodological tools. For the first time, quantitative data on stoichiometry and kinetics were collected and evaluated on different carbon sources. The results on various substrates were interpreted, with proposed metabolic mechanisms, by linking to the reported primary carbon metabolism of extreme halophilic Archaea. Moreover, results of chemostat cultures demonstrated that extreme halophilic organisms show Monod-kinetics on different sole carbon sources. A diauxic growth pattern was described on a mixture of substrates in batch cultivations. In addition, the methodologies presented here enable one to characterize the utilized strain Haloferax mediterranei (HFX) as a potential new host organism. Thus, this study offers a strong methodological basis as well as a fundamental physiological assessment for

Bioreactor droplets from liposome-stabilized all-aqueous emulsions

Science.gov (United States)

Dewey, Daniel C.; Strulson, Christopher A.; Cacace, David N.; Bevilacqua, Philip C.; Keating, Christine D.

2014-08-01

Artificial bioreactors are desirable for in vitro biochemical studies and as protocells. A key challenge is maintaining a favourable internal environment while allowing substrate entry and product departure. We show that semipermeable, size-controlled bioreactors with aqueous, macromolecularly crowded interiors can be assembled by liposome stabilization of an all-aqueous emulsion. Dextran-rich aqueous droplets are dispersed in a continuous polyethylene glycol (PEG)-rich aqueous phase, with coalescence inhibited by adsorbed ~130-nm diameter liposomes. Fluorescence recovery after photobleaching and dynamic light scattering data indicate that the liposomes, which are PEGylated and negatively charged, remain intact at the interface for extended time. Inter-droplet repulsion provides electrostatic stabilization of the emulsion, with droplet coalescence prevented even for submonolayer interfacial coatings. RNA and DNA can enter and exit aqueous droplets by diffusion, with final concentrations dictated by partitioning. The capacity to serve as microscale bioreactors is established by demonstrating a ribozyme cleavage reaction within the liposome-coated droplets.

Developing 3D microstructures for tissue engineering

DEFF Research Database (Denmark)

Mohanty, Soumyaranjan

casting process to generate various large scale tissue engineering constructs with single pore geometry with the desired mechanical stiffness and porosity. In addition, a new technique was developed to fa bricate dual-pore scaffolds for various tissue-engineering applications where 3D printing...... materials have been developed and tested for enhancing the differentiation of hiPSC-derived hepatocytes and fabricating biodegradable scaffolds for in-vivo tissue engineering applications. Along with various scaffolds fabrication methods we finally presented an optimized study of hepatic differentiation...... of hiPSC-derived DE cells cultured for 25 days in a 3D perfusion bioreactor system with an array of 16 small-scale tissue-bioreactors with integrated dual-pore pore scaffolds and flow rates. Hepatic differentiation and functionality of hiPSC-derived hepatocytes were successfully assessed and compared...

Intra-Arterial MR Perfusion Imaging of Meningiomas: Comparison to Digital Subtraction Angiography and Intravenous MR Perfusion Imaging.

Directory of Open Access Journals (Sweden)

Mark A Lum

Full Text Available To evaluate the ability of IA MR perfusion to characterize meningioma blood supply.Studies were performed in a suite comprised of an x-ray angiography unit and 1.5T MR scanner that permitted intraprocedural patient movement between the imaging modalities. Patients underwent intra-arterial (IA and intravenous (IV T2* dynamic susceptibility MR perfusion immediately prior to meningioma embolization. Regional tumor arterial supply was characterized by digital subtraction angiography and classified as external carotid artery (ECA dural, internal carotid artery (ICA dural, or pial. MR perfusion data regions of interest (ROIs were analyzed in regions with different vascular supply to extract peak height, full-width at half-maximum (FWHM, relative cerebral blood flow (rCBF, relative cerebral blood volume (rCBV, and mean transit time (MTT. Linear mixed modeling was used to identify perfusion curve parameter differences for each ROI for IA and IV MR imaging techniques. IA vs. IV perfusion parameters were also directly compared for each ROI using linear mixed modeling.18 ROIs were analyzed in 12 patients. Arterial supply was identified as ECA dural (n = 11, ICA dural (n = 4, or pial (n = 3. FWHM, rCBV, and rCBF showed statistically significant differences between ROIs for IA MR perfusion. Peak Height and FWHM showed statistically significant differences between ROIs for IV MR perfusion. RCBV and MTT were significantly lower for IA perfusion in the Dural ECA compared to IV perfusion. Relative CBF in IA MR was found to be significantly higher in the Dural ICA region and MTT significantly lower compared to IV perfusion.

Pulmonary perfusion ''without ventilation''

International Nuclear Information System (INIS)

Chapman, C.N.; Sziklas, J.J.; Spencer, R.P.; Rosenberg, R.J.

1983-01-01

An 88-yr-old man, with prior left upper lobectomy and phrenic nerve injury, had a ventilation/perfusion lung image. Both wash-in and equilibrium ventilation images showed no radioactive gas in the left lung. Nevertheless, the left lung was perfused. A similar result was obtained on a repeat study 8 days later. Delayed images, during washout, showed some radioactive gas in the left lung. Nearly absent ventilation (but continued perfusion) of that lung might have been related to altered gas dynamics brought about by the prior lobectomy, a submucosal bronchial lesion, phrenic nerve damage, and limited motion of the left part of the diaphragm. This case raises the issue of the degree of ventilation (and the phase relationship between the lungs) required for the entry of radioactive gas into a diseased lung, and the production of a ''reversed ventilation/perfusion mismatch.''

Long-term load duration induces N-cadherin down-regulation and loss of cell phenotype of nucleus pulposus cells in a disc bioreactor culture.

Science.gov (United States)

Li, Pei; Zhang, Ruijie; Wang, Liyuan; Gan, Yibo; Xu, Yuan; Song, Lei; Luo, Lei; Zhao, Chen; Zhang, Chengmin; Ouyang, Bin; Tu, Bing; Zhou, Qiang

2017-04-30

Long-term exposure to a mechanical load causes degenerative changes in the disc nucleus pulposus (NP) tissue. A previous study demonstrated that N-cadherin (N-CDH)-mediated signalling can preserve the NP cell phenotype. However, N-CDH expression and the resulting phenotype alteration in NP cells under mechanical compression remain unclear. The present study investigated the effects of the compressive duration on N-CDH expression and on the phenotype of NP cells in an ex vivo disc organ culture. Porcine discs were organ cultured in a self-developed mechanically active bioreactor for 7 days. The discs were subjected to different dynamic compression durations (1 and 8 h at a magnitude of 0.4 MPa and frequency of 1.0 Hz) once per day. Discs that were not compressed were used as controls. The results showed that long-term compression duration (8 h) significantly down-regulated the expression of N-CDH and NP-specific molecule markers (Brachyury, Laminin, Glypican-3 and Keratin 19), attenuated Alcian Blue staining intensity, decreased glycosaminoglycan (GAG) and hydroxyproline (HYP) contents and decreased matrix macromolecule (aggrecan and collagen II) expression compared with the short-term compression duration (1 h). Taken together, these findings demonstrate that long-term load duration can induce N-CDH down-regulation, loss of normal cell phenotype and result in attenuation of NP-related matrix synthesis in NP cells. © 2017 The Author(s).

Growth factors mediated differentiation of mesenchymal stem cells to cardiac polymicrotissue using hanging drop and bioreactor.

Science.gov (United States)

Konstantinou, Dimitrios; Lei, Ming; Xia, Zhidao; Kanamarlapudi, Venkateswarlu

2015-04-01

Heart disease is the major leading cause of death worldwide and the use of stem cells promises new ways for its treatment. The relatively easy and quick acquisition of human umbilical cord matrix mesenchymal stem cells (HUMSCs) and their properties make them useful for the treatment of cardiac diseases. Therefore, the main aim of this investigation was to create cardiac polymicrotissue from HUMSCs using a combination of growth factors [sphingosine-1-phosphate (S1P) and suramin] and techniques (hanging drop and bioreactor). Using designated culture conditions of the growth factors (100 nM S1P and 500 µM suramin), cardiomyocyte differentiation medium (CDM), hanging drop, bioreactor and differentiation for 7 days, a potential specific cardiac polymicrotissue was derived from HUMSCs. The effectiveness of growth factors alone or in combination in differentiation of HUMSCs to cardiac polymicrotissue was analysed by assessing the presence of cardiac markers by immunocytochemistry. This analysis demonstrated the importance of those growth factors for the differentiation. This study for the first time demonstrated the formation of a cardiac polymicrotissue under specific culture conditions. The polymicrotissue thus obtained may be used in future as a 'patch' to cover the injured cardiac region and would thereby be useful for the treatment of heart diseases. © 2014 International Federation for Cell Biology.

A study of extracellular matrix remodeling in aortic heart valves using a novel biaxial stretch bioreactor.

Science.gov (United States)

Lei, Ying; Masjedi, Shirin; Ferdous, Zannatul

2017-11-01

In aortic valves, biaxial cyclic stretch is known to modulate cell differentiation, extracellular matrix (ECM) synthesis and organization. We designed a novel bioreactor that can apply independent and precise stretch along radial and circumferential directions in a tissue culture environment. While this bioreactor can be used for either native or engineered tissues, this study determined matrix remodeling and strain distribution of aortic cusps after culturing under biaxial stretch for 14 days. The contents of collagen and glycosaminoglycans were determined using standard biochemical assays and compared with fresh controls. Strain fields in static cusps were more uniform than those in stretched cusps, which indicated degradation of the ECM fibers. The glycosaminoglycan content was significantly elevated in the static control as compared to fresh or stretched cusps, but no difference was observed in collagen content among the groups. The strain profile of freshly isolated fibrosa vs. ventricularis and left, right, and noncoronary cusps were also determined by Digital Image Correlation technique. Distinct strain patterns were observed under stretch on fibrosa and ventricularis sides and among the three cusps. This work highlights the critical role of the anisotropic ECM structure for proper functions of native aortic valves and the beneficial effects of biaxial stretch for maintenance of the native ECM structure. Copyright © 2017 Elsevier Ltd. All rights reserved.

Exploitation of Trametes versicolor for bioremediation of endocrine disrupting chemicals in bioreactors.

Directory of Open Access Journals (Sweden)

Cinzia Pezzella

Full Text Available Endocrine disrupting chemicals (EDCs are environmental contaminants causing increasing concerns due to their toxicity, persistence and ubiquity. In the present study, degradative capabilities of Trametes versicolor, Pleurotus ostreatus and Phanerochaete chrysosporium to act on five EDCs, which represent different classes of chemicals (phenols, parabens and phthalate and were first applied as single compounds, were assessed. T. versicolor was selected due to its efficiency against target EDCs and its potentialities were exploited against a mixture of EDCs in a cost-effective bioremediation process. A fed-batch approach as well as a starvation strategy were applied in order to reduce the need for input of 'fresh' biomass, and avoid the requirement for external nutrients. The fungus was successfully operated in two different bioreactors over one week. Semi-batch cultures were carried out by daily adding a mixture of EDCs to the bioreactors in a total of five consecutive degradation cycles. T. versicolor was able to efficiently remove all compounds during each cycle converting up to 21 mg L-1 day-1 of the tested EDCs. The maintained ability of T. versicolor to remove EDCs without any additional nutrients represents the main outcome of this study, which enables to forecast its application in a water treatment process.

Quantitative analysis of microbial biomass yield in aerobic bioreactor.

Science.gov (United States)

Watanabe, Osamu; Isoda, Satoru

2013-12-01

We have studied the integrated model of reaction rate equations with thermal energy balance in aerobic bioreactor for food waste decomposition and showed that the integrated model has the capability both of monitoring microbial activity in real time and of analyzing biodegradation kinetics and thermal-hydrodynamic properties. On the other hand, concerning microbial metabolism, it was known that balancing catabolic reactions with anabolic reactions in terms of energy and electron flow provides stoichiometric metabolic reactions and enables the estimation of microbial biomass yield (stoichiometric reaction model). We have studied a method for estimating real-time microbial biomass yield in the bioreactor during food waste decomposition by combining the integrated model with the stoichiometric reaction model. As a result, it was found that the time course of microbial biomass yield in the bioreactor during decomposition can be evaluated using the operational data of the bioreactor (weight of input food waste and bed temperature) by the combined model. The combined model can be applied to manage a food waste decomposition not only for controlling system operation to keep microbial activity stable, but also for producing value-added products such as compost on optimum condition. Copyright © 2013 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.

Hepatic perfusion changes in an experimental model of acute pancreatitis: Evaluation by perfusion CT

International Nuclear Information System (INIS)

Tutcu, Semra; Serter, Selim; Kaya, Yavuz; Kara, Eray; Nese, Nalan; Pekindil, Goekhan; Coskun, Teoman

2010-01-01

Purpose: It is known that acute pancreatitis may cause secondary changes in several organs. Liver is one of these involved organs. In different experimental studies hepatic damages were shown histopathologically in acute pancreatitis but there are a few studies about perfusion disorders that accompany these histopathologic changes. Perfusion CT (pCT) provides the ability to detect regional and global alterations in organ blood flow. The purpose of the study was to describe hepatic perfusion changes in experimental acute pancreatitis model with pCT. Materials and methods: Forty Sprague-Dawley rats of both genders with average weights of 250 g were used. Rats were randomized into two groups. Twenty rats were in control group and 20 in acute pancreatitis group. pCT was performed. Perfusion maps were formed by processing the obtained images with perfusion CT software. Blood flow (BF) and blood volume (BV) values were obtained from these maps. All pancreatic and liver tissues were taken off with laparotomy and histopathologic investigation was performed. Student's t test was used for statistical analyses. Results: In pCT we found statistically significant increase in blood volume in both lobes of liver and in blood flow in right lobe of the liver (p < 0.01). Although blood flow in left lobe of the liver increased, it did not reach statistical significance. Conclusion: The quantitative analysis of liver parenchyma with pCT showed that acute pancreatitis causes a significant perfusion changes in the hepatic tissue. Systemic mediators seem to be effective as well as local inflammatory changes in perfusion changes.

Hepatic perfusion changes in an experimental model of acute pancreatitis: Evaluation by perfusion CT

Energy Technology Data Exchange (ETDEWEB)

Tutcu, Semra [Department of Surgery, Celal Bayar University, School of Medicine, Manisa (Turkey); Serter, Selim, E-mail: serterselim@gmail.co [Department of Radiology, Celal Bayar University, School of Medicine, Manisa (Turkey); Kaya, Yavuz; Kara, Eray [Department of Surgery, Celal Bayar University, School of Medicine, Manisa (Turkey); Nese, Nalan [Department of Pathology, Celal Bayar University, School of Medicine, Manisa (Turkey); Pekindil, Goekhan [Department of Radiology, Celal Bayar University, School of Medicine, Manisa (Turkey); Coskun, Teoman [Department of Surgery, Celal Bayar University, School of Medicine, Manisa (Turkey)

2010-08-15

Purpose: It is known that acute pancreatitis may cause secondary changes in several organs. Liver is one of these involved organs. In different experimental studies hepatic damages were shown histopathologically in acute pancreatitis but there are a few studies about perfusion disorders that accompany these histopathologic changes. Perfusion CT (pCT) provides the ability to detect regional and global alterations in organ blood flow. The purpose of the study was to describe hepatic perfusion changes in experimental acute pancreatitis model with pCT. Materials and methods: Forty Sprague-Dawley rats of both genders with average weights of 250 g were used. Rats were randomized into two groups. Twenty rats were in control group and 20 in acute pancreatitis group. pCT was performed. Perfusion maps were formed by processing the obtained images with perfusion CT software. Blood flow (BF) and blood volume (BV) values were obtained from these maps. All pancreatic and liver tissues were taken off with laparotomy and histopathologic investigation was performed. Student's t test was used for statistical analyses. Results: In pCT we found statistically significant increase in blood volume in both lobes of liver and in blood flow in right lobe of the liver (p < 0.01). Although blood flow in left lobe of the liver increased, it did not reach statistical significance. Conclusion: The quantitative analysis of liver parenchyma with pCT showed that acute pancreatitis causes a significant perfusion changes in the hepatic tissue. Systemic mediators seem to be effective as well as local inflammatory changes in perfusion changes.

A novel process-based model of microbial growth: self-inhibition in Saccharomyces cerevisiae aerobic fed-batch cultures.

Science.gov (United States)

Mazzoleni, Stefano; Landi, Carmine; Cartenì, Fabrizio; de Alteriis, Elisabetta; Giannino, Francesco; Paciello, Lucia; Parascandola, Palma

2015-07-30

Microbial population dynamics in bioreactors depend on both nutrients availability and changes in the growth environment. Research is still ongoing on the optimization of bioreactor yields focusing on the increase of the maximum achievable cell density. A new process-based model is proposed to describe the aerobic growth of Saccharomyces cerevisiae cultured on glucose as carbon and energy source. The model considers the main metabolic routes of glucose assimilation (fermentation to ethanol and respiration) and the occurrence of inhibition due to the accumulation of both ethanol and other self-produced toxic compounds in the medium. Model simulations reproduced data from classic and new experiments of yeast growth in batch and fed-batch cultures. Model and experimental results showed that the growth decline observed in prolonged fed-batch cultures had to be ascribed to self-produced inhibitory compounds other than ethanol. The presented results clarify the dynamics of microbial growth under different feeding conditions and highlight the relevance of the negative feedback by self-produced inhibitory compounds on the maximum cell densities achieved in a bioreactor.

Biotic transformation of anticoccidials in soil using a lab-scale bio-reactor as a precursor-tool

DEFF Research Database (Denmark)

Hansen, Martin; Björklund, Erland; Krogh, Kristine A

2012-01-01

incubated for 200 h with a mixed culture of soil bacteria. Samples were analyzed by LC-MS/MS and potential transformation products were tentatively identified. Salinomycin was degraded under aerobic conditions and traces could be found after 200 h, however, seems more persistent under anaerobic conditions....... Four transformation products of salinomycin were discovered. Robenidine was degraded under aerobic and anaerobic conditions, however, traces of robenidine were observed after 200 h. Five biotic transformation products of robenidine were discovered.......Two anticoccidial agents, salinomycin and robenidine, heavily used in the worldwide veterinary meat production, were investigated for their potential biotic degradation by cultured soil bacteria. The degradation-study was performed in lab-scale bio-reactors under aerobic and anaerobic conditions...

Contralateral thalamic hypoperfusion on brain perfusion SPECT

International Nuclear Information System (INIS)

Lee, Seok Mo; Bae, Sang Kyun; Yoo, Kyung Moo; Yum, Ha Yong

2000-01-01

Brain perfusion single photon emission computed tomography (SPECT) is useful for the localization of cerebrovascular lesion and sometimes reveals more definite lesion than radiologic imaging modality such as CT or MRI does. The purpose of this study was to evaluate the diagnostic usefulness of brain perfusion SPECT in patients with hemisensory impairment. Thirteen consecutive patients (M:F= 8:5, mean age = 48) who has hemisensory impairment were included. Brain perfusion SPECT was performed after intravenous injection of 1110 MBq of Tc-99m ECD. The images were obtained using a dual-head gamma camera with ultra-high resolution collimator. Semiquantitative analysis was performed after placing multiple ROIs on cerebral cortex, basal ganglia, thalamus and cerebellum. There were 10 patients with left hemisensory impairment and 3 patients with right-sided symptom. Only 2 patients revealed abnormal signal change in the thalamus on MRI. But brain perfusion SPECT showed decreased perfusion in the thalamus in 9 patients. Six patients among 10 patients with left hemisensory impairment revealed decreased perfusion in the contralateral thalamus on brain SPECT. The other 4 patients revealed no abnormality. Two patients among 3 patients with right hemisensory impairment also showed decreased perfusion in the contralateral thalamus on brain SPECT. One patients with right hemisensory impairment showed ipsilateral perfusion decrease. Two patients who had follow-up brain perfusion SEPCT after treatment revealed normalization of perfusion in the thalamus. Brain perfusion SPECT might be a useful tool in diagnosing patients with hemisensory impairment

Magnetic resonance perfusion imaging without contrast media

International Nuclear Information System (INIS)

Martirosian, Petros; Graf, Hansjoerg; Schick, Fritz; Boss, Andreas; Schraml, Christina; Schwenzer, Nina F.; Claussen, Claus D.

2010-01-01

Principles of magnetic resonance imaging techniques providing perfusion-related contrast weighting without administration of contrast media are reported and analysed systematically. Especially common approaches to arterial spin labelling (ASL) perfusion imaging allowing quantitative assessment of specific perfusion rates are described in detail. The potential of ASL for perfusion imaging was tested in several types of tissue. After a systematic comparison of technical aspects of continuous and pulsed ASL techniques the standard kinetic model and tissue properties of influence to quantitative measurements of perfusion are reported. For the applications demonstrated in this paper a flow-sensitive alternating inversion recovery (FAIR) ASL perfusion preparation approach followed by true fast imaging with steady precession (true FISP) data recording was developed and implemented on whole-body scanners operating at 0.2, 1.5 and 3 T for quantitative perfusion measurement in various types of tissue. ASL imaging provides a non-invasive tool for assessment of tissue perfusion rates in vivo. Images recorded from kidney, lung, brain, salivary gland and thyroid gland provide a spatial resolution of a few millimetres and sufficient signal to noise ratio in perfusion maps after 2-5 min of examination time. Newly developed ASL techniques provide especially high image quality and quantitative perfusion maps in tissues with relatively high perfusion rates (as also present in many tumours). Averaging of acquisitions and image subtraction procedures are mandatory, leading to the necessity of synchronization of data recording to breathing in abdominal and thoracic organs. (orig.)

Hepatic arterial perfusion increases in the early stage of severe acute pancreatitis patients: Evaluation by perfusion computed tomography

International Nuclear Information System (INIS)

Koyasu, Sho; Isoda, Hiroyoshi; Tsuji, Yoshihisa; Yamamoto, Hiroshi; Matsueda, Kazuhiro; Watanabe, Yuji; Chiba, Tsutomu; Togashi, Kaori

2012-01-01

Purpose: Although hepatic perfusion abnormalities have been reported in patients with acute pancreatitis, hepatic perfusion with severe acute pancreatitis (SAP) has not been quantitatively evaluated in humans. Therefore, we investigated hepatic perfusion in patients with SAP using perfusion CT. Materials and methods: Hepatic perfusion CT was performed in 67 patients with SAP within 3 days after symptom onset. The patients were diagnosed as having SAP according to the Atlanta criteria. Fifteen cases were established as a control group. Perfusion CT was obtained for 54 s beginning with a bolus injection of 40 ml of contrast agent (600–630 mgI/kg) at a flow rate of 4 ml/s. Perfusion data were analyzed by the dual-input maximum slope method to obtain hepatic arterial perfusion (HAP) and hepatic portal perfusion (HPP). Finally, we compared HAP and HPP in SAP patients with those in the control group, respectively. Results: Average HAP was significantly higher in SAP patients than in the control group (75.1 ± 38.0 vs. 38.2 ± 9.0 ml/min/100 ml; p < 0.001). There was no significant difference in average HPP between SAP patients and the control group (206.7 ± 54.9 vs. 204.4 ± 38.5 ml/min/100 ml; p = 0.92). Conclusion: Using quantitative analysis on perfusion CT, we first demonstrated an increase of HAP in the right hepatic lobe in SAP patients.

Intelligent Bioreactor Management Information System (IBM-IS) for Mitigation of Greenhouse Gas Emissions

Energy Technology Data Exchange (ETDEWEB)

Paul Imhoff; Ramin Yazdani; Don Augenstein; Harold Bentley; Pei Chiu

2010-04-30

Methane is an important contributor to global warming with a total climate forcing estimated to be close to 20% that of carbon dioxide (CO2) over the past two decades. The largest anthropogenic source of methane in the US is 'conventional' landfills, which account for over 30% of anthropogenic emissions. While controlling greenhouse gas emissions must necessarily focus on large CO2 sources, attention to reducing CH4 emissions from landfills can result in significant reductions in greenhouse gas emissions at low cost. For example, the use of 'controlled' or bioreactor landfilling has been estimated to reduce annual US greenhouse emissions by about 15-30 million tons of CO2 carbon (equivalent) at costs between $3-13/ton carbon. In this project we developed or advanced new management approaches, landfill designs, and landfill operating procedures for bioreactor landfills. These advances are needed to address lingering concerns about bioreactor landfills (e.g., efficient collection of increased CH4 generation) in the waste management industry, concerns that hamper bioreactor implementation and the consequent reductions in CH4 emissions. Collectively, the advances described in this report should result in better control of bioreactor landfills and reductions in CH4 emissions. Several advances are important components of an Intelligent Bioreactor Management Information System (IBM-IS).

Improved visualization of delayed perfusion in lung MRI

International Nuclear Information System (INIS)

Risse, Frank; Eichinger, Monika; Kauczor, Hans-Ulrich; Semmler, Wolfhard; Puderbach, Michael

2011-01-01

Introduction: The investigation of pulmonary perfusion by three-dimensional (3D) dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) was proposed recently. Subtraction images are generated for clinical evaluation, but temporal information is lost and perfusion defects might therefore be masked in this process. The aim of this study is to demonstrate a simple analysis strategy and classification for 3D-DCE-MRI perfusion datasets in the lung without omitting the temporal information. Materials and methods: Pulmonary perfusion measurements were performed in patients with different lung diseases using a 1.5 T MR-scanner with a time-resolved 3D-GRE pulse sequence. 25 3D-volumes were acquired after iv-injection of 0.1 mmol/kg KG Gadolinium-DTPA. Three parameters were determined for each pixel: (1) peak enhancement S n,max normalized to the arterial input function to detect regions of reduced perfusion; (2) time between arterial peak enhancement in the large pulmonary artery and tissue peak enhancement τ to visualize regions with delayed bolus onset; and (3) ratio R = S n,max /τ was calculated to visualize impaired perfusion, irrespectively of whether related to reduced or delayed perfusion. Results: A manual selection of peak perfusion images is not required. Five different types of perfusion can be found: (1) normal perfusion; (2) delayed non-reduced perfusion; (3) reduced non-delayed perfusion; (4) reduced and delayed perfusion; and (5) no perfusion. Types II and IV could not be seen in subtraction images since the temporal information is necessary for this purpose. Conclusions: The analysis strategy in this study allows for a simple and observer-independent visualization and classification of impaired perfusion in dynamic contrast-enhanced pulmonary perfusion MRI by using the temporal information of the datasets.

Use of NASA Bioreactor in Engineering Tissue for Bone Repair

Science.gov (United States)

Duke, Pauline

1998-01-01

This study was proposed in search for a new alternative for bone replacement or repair. Because the systems commonly used in repair of bony defects form bone by going through a cartilaginous phase, implantation of a piece of cartilage could enhance the healing process by having a more advanced starting point. However, cartilage has seldom been used to replace bone due, in part, to the limitations in conventional culture systems that did not allow production of enough tissue for implants. The NASA-developed bioreactors known as STLV (Slow Turning Lateral Vessel) provide homogeneous distribution of cells, nutrients, and waste products, with less damaging turbulence and shear forces than conventional systems. Cultures under these conditions have higher growth rates, viability, and longevity, allowing larger "tissue-like" aggregates to form, thus opening the possibilities of producing enough tissue for implantation, along with the inherent advantages of in vitro manipulations. To assure large numbers of cells and to eliminate the use of timed embryos, we proposed to use an immortalized mouse limb bud cell line as the source of cells.

Ventilation-perfused studies using SPECT

International Nuclear Information System (INIS)

Zwijnenburg, A.

1989-01-01

A method for the quantitative analysis of ventilation-perfusion SPECT studies is decribed and an effort is made to evaluate its usefullness. The technical details of the emthod are described. In the the transaxial reconstructions of the tomographic studies the contour of the lungs is detected and regional values of lung volume, ventilation, perfusion and ventilation-perfusion ratios are calculated. The method is operator independent. The lung volume calculations from the SPECT studies are validated by comparing them with lung volume measurements using the helium dilution technique. A good correlation (r=0.91) was found between the two volumes. SPECT volume was greater than the volume measured with helium dilution, which was attributed to non-gas-containing structures in the. lungs. The use of ventilation-perfusion ratio SPECT is described to evaluate the effect of ionizing radiation on the lungs in patients treated with mantle field irradiation for Hodgkin's disease. Perfusion changes appear as early as 2 months after the start of irradiation. Ventilation changes appear later and relatively minor. No changes are seen outside the radiation portals. The ventilation-perfusion inequality in pulmonary sarcoidosis is treated. It is suggested that the decrease D LCO in these patients may be partly due to an even distribution of ventilation perfusion ratios. An effort is made to establish the properties of a new tracer used for the assessment of the metabolic function of the pulmonary endothelium. The lung uptake of I-123 IMP mimics the distribution of a perfusion tracer and it is suggested that this tracer may be useful for the early detection of pulmonary vascular damage, even when blood flow is still intact. Some aspects of the use of Kr-81m as a ventilation tracer are discussed as well as the effect of noise on Kr-81m SPECT reconstructions. (author). 146 refs.; 39 figs.; 8 tabs

Automatic assessment of cardiac perfusion MRI

DEFF Research Database (Denmark)

Ólafsdóttir, Hildur; Stegmann, Mikkel Bille; Larsson, Henrik B.W.

2004-01-01

In this paper, a method based on Active Appearance Models (AAM) is applied for automatic registration of myocardial perfusion MRI. A semi-quantitative perfusion assessment of the registered image sequences is presented. This includes the formation of perfusion maps for three parameters; maximum up...

Perchlorate remediation using packed-bed bioreactors and electricity generation in microbial fuel cells (MFCs)

Science.gov (United States)

Min, Booki

Two pilot-scale fixed bed bioreactors were operated in continuous mode in order to treat groundwater contaminated by perchlorate. The bioreactors were constructed and operated side-by-side at the Texas Street Well Facility in Redlands, California. Each reactor was packed with either sand or plastic media. A perchlorate-reducing bacterium, Dechlorosoma sp. KJ, was used to inoculate the bioreactors. Perchlorate was successfully removed down to a non-detectable level (microbial fuel cells (MFCs), which were run either in batch or continuous mode. In batch experiments, both a pure culture (Geobactor metallireducens) and a mixed culture (wastewater inoculum) were used as the biocatalyst, and acetate was added as substrate in the anode chamber of the MFC. Power output in a membrane MFC with either inoculum was essentially the same, with 40 +/- 1 mW/m2 for G. metallireducens and 38 +/- 1 mW/m2 for mixed culture. A different type of the MFC containing a salt bridge instead of a membrane system was examined to generate power using the same substrate and pure culture as used in the membrane MFC. Power output in the salt bridge MFC was 2.2 mW/m 2. It was found that the lower power output was directly attributed to the higher internal resistance of the salt bridge system (19920 +/- 50 O) in comparison with that of the membrane system (1286 +/- 1 O). Continuous electricity generation was examined in a flat plate microbial fuel cell (FPMFC) using domestic wastewater and specific organic substrates. The FPMFC, containing a combined electrode/proton exchange membrane (PEM), was initially acclimated for one month to domestic wastewater, and then was operated as a plug flow reactor system. Power density using domestic wastewater as a substrate was 72 +/- 1 mW/m2 at a liquid flow rate of 0.39 mL/min (1.1 hr hydraulic retention time, HRT), and COD removal was 42%. At a longer HRT of 4.0 hr, the COD removal increased to 79%, and power density was 43 mW/m2. Several organic compounds

Can preoperative myocardial perfusion scintigraphy predict changes in left ventricular perfusion and function after coronary artery bypass graft surgery?

DEFF Research Database (Denmark)

Eckardt, Rozy; Kjeldsen, Bo Juel; Johansen, Allan

2012-01-01

OBJECTIVESWe wanted to evaluate whether preoperative myocardial perfusion scintigraphy (MPS) could predict changes in cardiac symptoms and postoperative myocardial perfusion and left ventricular function after coronary artery bypass grafting (CABG).METHODSNinety-two patients with stable angina...... in 26%. Left ventricular ejection fraction (LVEF), which was normal before operation in 45%, improved in 40% of all patients. The increase in LVEF was not related to the preoperative pattern of perfusion defects. Of 30 patients with normalized perfusion after CABG, 29 (97%) had reversible defects...... that reversible or partly reversible perfusion defects at a preoperative MPS have a high chance of normalized myocardial perfusion assessed by MPS 6 months after operation. Normal perfusion is obtained almost exclusively in territories with reversible ischaemia. Symptoms improved in nearly all patients and LVEF...

Thinking beyond the Bioreactor Box: Incorporating Stream Ecology into Edge-of-Field Nitrate Management.

Science.gov (United States)

Goeller, Brandon C; Febria, Catherine M; Harding, Jon S; McIntosh, Angus R

2016-05-01

Around the world, artificially drained agricultural lands are significant sources of reactive nitrogen to stream ecosystems, creating substantial stream health problems. One management strategy is the deployment of denitrification enhancement tools. Here, we evaluate the factors affecting the potential of denitrifying bioreactors to improve stream health and ecosystem services. The performance of bioreactors and the structure and functioning of stream biotic communities are linked by environmental parameters like dissolved oxygen and nitrate-nitrogen concentrations, dissolved organic carbon availability, flow and temperature regimes, and fine sediment accumulations. However, evidence of bioreactors' ability to improve waterway health and ecosystem services is lacking. To improve the potential of bioreactors to enhance desirable stream ecosystem functioning, future assessments of field-scale bioreactors should evaluate the influences of bioreactor performance on ecological indicators such as primary production, organic matter processing, stream metabolism, and invertebrate and fish assemblage structure and function. These stream health impact assessments should be conducted at ecologically relevant spatial and temporal scales. Bioreactors have great potential to make significant contributions to improving water quality, stream health, and ecosystem services if they are tailored to site-specific conditions and implemented strategically with land-based and stream-based mitigation tools within watersheds. This will involve combining economic, logistical, and ecological information in their implementation. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

Cancer cells growing on perfused 3D collagen model produced higher reactive oxygen species level and were more resistant to cisplatin compared to the 2D model.

Science.gov (United States)

Liu, Qingxi; Zhang, Zijiang; Liu, Yupeng; Cui, Zhanfeng; Zhang, Tongcun; Li, Zhaohui; Ma, Wenjian

2018-03-01

Three-dimensional (3D) collagen scaffold models, due to their ability to mimic the tissue and organ structure in vivo, have received increasing interest in drug discovery and toxicity evaluation. In this study, we developed a perfused 3D model and studied cellular response to cytotoxic drugs in comparison with traditional 2D cell cultures as evaluated by cancer drug cisplatin. Cancer cells grown in perfused 3D environments showed increased levels of reactive oxygen species (ROS) production compared to the 2D culture. As determined by growth analysis, cells in the 3D culture, after forming a spheroid, were more resistant to the cancer drug cisplatin compared to that of the 2D cell culture. In addition, 3D culturing cells showed elevated level of ROS, indicating a physiological change or the formation of a microenvironment that resembles tumor cells in vivo. These data revealed that cellular response to drugs for cells growing in 3D environments are dramatically different from that of 2D cultured cells. Thus, the perfused 3D collagen scaffold model we report here might be a potentially very useful tool for drug analysis.

Application of dynamic membranes in anaerobic membranes in anaerobic membrane bioreactor systems

NARCIS (Netherlands)

Erşahin, M.E.

2015-01-01

Anaerobic membrane bioreactors (AnMBRs) physically ensure biomass retention by the application of a membrane filtration process. With growing application experiences from aerobic membrane bioreactors (MBRs), the combination of membrane and anaerobic processes has received much attention and become

A dual flow bioreactor with controlled mechanical stimulation for cartilage tissue engineering

NARCIS (Netherlands)

Spitters, Tim; Leijten, Jeroen Christianus Hermanus; Deus, F.D.; Costa, I.B.F.; van Apeldoorn, Aart A.; van Blitterswijk, Clemens; Karperien, Hermanus Bernardus Johannes

2013-01-01

In cartilage tissue engineering bioreactors can create a controlled environment to study chondrocyte behavior under mechanical stimulation or produce chondrogenic grafts of clinically relevant size. Here we present a novel bioreactor, which combines mechanical stimulation with a two compartment

Design and testing of a unique randomized gravity, continuous flow bioreactor

Science.gov (United States)

Lassiter, Carroll B.

1993-01-01

A rotating, null gravity simulator, or Couette bioreactor was successfully used for the culture of mammalian cells in a simulated microgravity environment. Two limited studies using Lipomyces starkeyi and Streptomyces clavuligerus were also conducted under conditions of simulated weightlessness. Although these studies with microorganisms showed promising preliminary results, oxygen limitations presented significant limitations in studying the biochemical and cultural characteristics of these cell types. Microbial cell systems such as bacteria and yeast promise significant potential as investigative models to study the effects of microgravity on membrane transport, as well as substrate induction of inactive enzyme systems. Additionally, the smaller size of the microorganisms should further reduce the gravity induced oscillatory particle motion and thereby improve the microgravity simulation on earth. Focus is on the unique conceptual design, and subsequent development of a rotating bioreactor that is compatible with the culture and investigation of microgravity effects on microbial systems. The new reactor design will allow testing of highly aerobic cell types under simulated microgravity conditions. The described reactor affords a mechanism for investigating the long term effects of reduced gravity on cellular respiration, membrane transfer, ion exchange, and substrate conversions. It offers the capability of dynamically altering nutrients, oxygenation, pH, carbon dioxide, and substrate concentration without disturbing the microgravity simulation, or Couette flow, of the reactor. All progeny of the original cell inoculum may be acclimated to the simulated microgravity in the absence of a substrate or nutrient. The reactor has the promise of allowing scientists to probe the long term effects of weightlessness on cell interactions in plants, bacteria, yeast, and fungi. The reactor is designed to have a flow field growth chamber with uniform shear stress, yet transfer

Norfloxacin degradation by Bacillus subtilis strains able to produce biosurfactants on a bioreactor scale

Directory of Open Access Journals (Sweden)

Jałowiecki Łukasz

2017-01-01

Full Text Available The discharge of antibiotics into the environment has become a major concern since this group of pharmaceuticals influence on microbial communities not only by its mode of action, but also because of the risk of a worldwide dispersal of antibiotic resistance genes (ARG. Antibiotics residues have been found in various environments such as waters, sediments, and soils. Moreover, most WWTPs are not designed to treat such kind of pollutants, which remain incompletely removed. Currently, biodegradation processes which involved bacterial strains with increased degradation capabilities is one of the most promising technique. The aim of this study was to evaluate the norfloxacin biodegradation potential of the three Bacillus subtilis strains named T-1, T’-1 and I’-1a on a bioreactor scale. The aerobic degradation was conducted in a 5-liter bioreactor on minimal salts medium in co-metabolic culture supplemented with glucose. The degradation rate of norfloxacin was determined with the HPLC technique. The surface tension was determined using ring method in order to observe the changes in biosurfactants production. Also, the biofilm formation abilities of the bacteria with two quantitative methods, crystal violet (CV method and TTC-based test and enzymes production were evaluated.

Ex-vivo machine perfusion for kidney preservation.

Science.gov (United States)

Hamar, Matyas; Selzner, Markus

2018-06-01

Machine perfusion is a novel strategy to decrease preservation injury, improve graft assessment, and increase organ acceptance for transplantation. This review summarizes the current advances in ex-vivo machine-based kidney preservation technologies over the last year. Ex-vivo perfusion technologies, such as hypothermic and normothermic machine perfusion and controlled oxygenated rewarming, have gained high interest in the field of organ preservation. Keeping kidney grafts functionally and metabolically active during the preservation period offers a unique chance for viability assessment, reconditioning, and organ repair. Normothermic ex-vivo kidney perfusion has been recently translated into clinical practice. Preclinical results suggest that prolonged warm perfusion appears superior than a brief end-ischemic reconditioning in terms of renal function and injury. An established standardized protocol for continuous warm perfusion is still not available for human grafts. Ex-vivo machine perfusion represents a superior organ preservation method over static cold storage. There is still an urgent need for the optimization of the perfusion fluid and machine technology and to identify the optimal indication in kidney transplantation. Recent research is focusing on graft assessment and therapeutic strategies.

Simulation of three-phase fluidized bioreactors for denitrification

International Nuclear Information System (INIS)

Hamza, A.V.; Dolan, J.F.; Wong, E.W.

1981-03-01

Fluidized-bed bioreactors were developed and operated at three scales (diameters of 0.1, 0.2, and 0.5 m) by the Chemical Technology Division. The performance of these reactors in denitrification was simulated using the following modified form of Monod kinetics to describe the reaction kinetics: rate = V/sub max/ (NO 3 - /K/sub s/ + NO 3 - ) (% biomass). In the fluids-movement portion of the simulation the tanks-in-series approximation to backmixing was used. This approach yielded a V/sub max/ of 3.5 g/m 3 -min (% biomass) and a K/sub s/ of 163 g/m 3 for the 0.5-m bioreactor. Values of V/sub max/ and K/sub s/ were also determined for data derived from the 0.1-m bioreactor, but inadequate RTD data reduced the confidence level in these results. A complication in denitrification is the multi-step nature of the reduction from nitrate to nitrite to hyponitrite and finally to nitrogen. An experimental study of the effect of biomass loading upon denitrification was begun. It is recommended that the experimental work be continued

Cultivation of methanogenic community from 2-km deep subseafloor coalbeds using a continuous-flow bioreactor

Science.gov (United States)

Imachi, H.; Tasumi, E.; Morono, Y.; Ito, M.; Takai, K.; Inagaki, F.

2013-12-01

Deep subseafloor environments associated with hydrocarbon reservoirs have been least explored by previous scientific drilling and hence the nature of deep subseafloor life and its ecological roles in the carbon cycle remain largely unknown. In this study, we performed cultivation of subseafloor methanogenic communities using a continuous-flow bioreactor with polyurethane sponges, called down-flow hanging sponge (DHS) reactor. The sample used for the reactor cultivation was obtained from 2 km-deep coalbeds off the Shimokita Peninsula of Japan, the northwestern Pacific, during the Integrated Ocean Drilling Program (IODP) Expedition 337 using a riser drilling technology of the drilling vessel Chikyu. The coalbed samples were incubated anaerobically in the DHS reactor at the in-situ temperature of 40°C. Synthetic seawater supplemented with a tiny amount of yeast extract, acetate, propionate and butyrate was provided into the DHS reactor. After 34 days of the bioreactor operation, a small production of methane was observed. The methane concentration was gradually increased and the stable carbon isotopic composition of methane was consistency 13C-depleted during the bioreactor operation, indicating the occurrence of microbial methanogenesis. Microscopic observation showed that the enrichment culture contained a variety of microorganisms, including methanogen-like rod-shaped cells with F420 auto-fluorescence. Interestingly, many spore-like particles were observed in the bioreactor enrichment. Phylogenetic analysis of 16S rRNA genes showed the growth of phylogenetically diverse bacteria and archaea in the DHS reactor. Predominant archaeal components were closely related to hydrogenotrophic methanogens within the genus Methanobacterium. Some predominant bacteria were related to the spore-formers within the class Clostridia, which are overall in good agreement with microscopic observations. By analyzing ion images using a nano-scale secondary ion mass spectrometry (Nano

Formation of three-dimensional cell/polymer constructs for bone tissue engineering in a spinner flask and a rotating wall vessel bioreactor

Science.gov (United States)

Sikavitsas, Vassilios I.; Bancroft, Gregory N.; Mikos, Antonios G.; McIntire, L. V. (Principal Investigator)

2002-01-01

The aim of this study is to investigate the effect of the cell culture conditions of three-dimensional polymer scaffolds seeded with rat marrow stromal cells (MSCs) cultured in different bioreactors concerning the ability of these cells to proliferate, differentiate towards the osteoblastic lineage, and generate mineralized extracellular matrix. MSCs harvested from male Sprague-Dawley rats were culture expanded, seeded on three-dimensional porous 75:25 poly(D,L-lactic-co-glycolic acid) biodegradable scaffolds, and cultured for 21 days under static conditions or in two model bioreactors (a spinner flask and a rotating wall vessel) that enhance mixing of the media and provide better nutrient transport to the seeded cells. The spinner flask culture demonstrated a 60% enhanced proliferation at the end of the first week when compared to static culture. On day 14, all cell/polymer constructs exhibited their maximum alkaline phosphatase activity (AP). Cell/polymer constructs cultured in the spinner flask had 2.4 times higher AP activity than constructs cultured under static conditions on day 14. The total osteocalcin (OC) secretion in the spinner flask culture was 3.5 times higher than the static culture, with a peak OC secretion occurring on day 18. No considerable AP activity and OC secretion were detected in the rotating wall vessel culture throughout the 21-day culture period. The spinner flask culture had the highest calcium content at day 14. On day 21, the calcium deposition in the spinner flask culture was 6.6 times higher than the static cultured constructs and over 30 times higher than the rotating wall vessel culture. Histological sections showed concentration of cells and mineralization at the exterior of the foams at day 21. This phenomenon may arise from the potential existence of nutrient concentration gradients at the interior of the scaffolds. The better mixing provided in the spinner flask, external to the outer surface of the scaffolds, may explain the

Introducing Textiles as Material of Construction of Ethanol Bioreactors

Directory of Open Access Journals (Sweden)

Osagie A. Osadolor

2014-11-01

Full Text Available The conventional materials for constructing bioreactors for ethanol production are stainless and cladded carbon steel because of the corrosive behaviour of the fermenting media. As an alternative and cheaper material of construction, a novel textile bioreactor was developed and examined. The textile, coated with several layers to withstand the pressure, resist the chemicals inside the reactor and to be gas-proof was welded to form a 30 L lab reactor. The reactor had excellent performance for fermentative production of bioethanol from sugar using baker’s yeast. Experiments with temperature and mixing as process parameters were performed. No bacterial contamination was observed. Bioethanol was produced for all conditions considered with the optimum fermentation time of 15 h and ethanol yield of 0.48 g/g sucrose. The need for mixing and temperature control can be eliminated. Using a textile bioreactor at room temperature of 22 °C without mixing required 2.5 times longer retention time to produce bioethanol than at 30 °C with mixing. This will reduce the fermentation investment cost by 26% for an ethanol plant with capacity of 100,000 m3 ethanol/y. Also, replacing one 1300 m3 stainless steel reactor with 1300 m3 of the textile bioreactor in this plant will reduce the fermentation investment cost by 19%.

Anaerobic digestion of citrus waste using two-stage membrane bioreactor

Science.gov (United States)

Millati, Ria; Lukitawesa; Dwi Permanasari, Ervina; Wulan Sari, Kartika; Nur Cahyanto, Muhammad; Niklasson, Claes; Taherzadeh, Mohammad J.

2018-03-01

Anaerobic digestion is a promising method to treat citrus waste. However, the presence of limonene in citrus waste inhibits anaerobic digestion process. Limonene is an antimicrobial compound and could inhibit methane forming bacteria that takes a longer time to recover than the injured acid forming bacteria. Hence, volatile fatty acids will be accumulated and methane production will be decreased. One way to solve this problem is by conducting anaerobic digestion process into two stages. The first step is aimed for hydrolysis, acidogenesis, and acetogenesis reactions and the second stage is aimed for methanogenesis reaction. The separation of the system would further allow each stage in their optimum conditions making the process more stable. In this research, anaerobic digestion was carried out in batch operations using 120 ml-glass bottle bioreactors in 2 stages. The first stage was performed in free-cells bioreactor, whereas the second stage was performed in both bioreactor of free cells and membrane bioreactor. In the first stage, the reactor was set into ‘anaerobic’ and ‘semi-aerobic’ conditions to examine the effect of oxygen on facultative anaerobic bacteria in acid production. In the second stage, the protection of membrane towards the cells against limonene was tested. For the first stage, the basal medium was prepared with 1.5 g VS of inoculum and 4.5 g VS of citrus waste. The digestion process was carried out at 55°C for four days. For the second stage, the membrane bioreactor was prepared with 3 g of cells that were encased and sealed in a 3×6 cm2 polyvinylidene fluoride membrane. The medium contained 40 ml basal medium and 10 ml liquid from the first stage. The bioreactors were incubated at 55°C for 2 days under anaerobic condition. The results from the first stage showed that the maximum total sugar under ‘anaerobic’ and ‘semi-aerobic’ conditions was 294.3 g/l and 244.7 g/l, respectively. The corresponding values for total volatile

Biogeochemistry of the compost bioreactor components of a composite acid mine drainage passive remediation system

International Nuclear Information System (INIS)

Johnson, D. Barrie; Hallberg, Kevin B.

2005-01-01

The compost bioreactor ('anaerobic cell') components of three composite passive remediation systems constructed to treat acid mine drainage (AMD) at the former Wheal Jane tin mine, Cornwall, UK were studied over a period of 16 months. While there was some amelioration of the preprocessed AMD in each of the three compost bioreactors, as evidenced by pH increase and decrease in metal concentrations, only one of the cells showed effective removal of the two dominant heavy metals (iron and zinc) present. With two of the compost bioreactors, concentrations of soluble (ferrous) iron draining the cells were significantly greater than those entering the reactors, indicating that there was net mobilisation (by reductive dissolution) of colloidal and/or solid-phase ferric iron compounds within the cells. Soluble sulfide was also detected in waters draining all three compost bioreactors which was rapidly oxidised, in contrast to ferrous iron. Oxidation and hydrolysis of iron, together with sulfide oxidation, resulted in reacidification of processed AMD downstream of the compost bioreactors in two of the passive treatment systems. The dominant cultivatable microorganism in waters draining the compost bioreactors was identified, via analysis of its 16S rRNA gene, as a Thiomonas sp. and was capable of accelerating the dissimilatory oxidation of both ferrous iron and reduced sulfur compounds. Sulfate-reducing bacteria (SRB) were also detected, although only in the bioreactor that was performing well were these present in significant numbers. This particular compost bioreactor had been shut down for 10 months prior to the monitoring period due to operational problems. This unforeseen event appears to have allowed more successful development of AMD-tolerant and other microbial populations with critical roles in AMD bioremediation, including neutrophilic SRB (nSRB), in this compost bioreactor than in the other two, where the throughput of AMD was not interrupted. This study has

Biogeochemistry of the compost bioreactor components of a composite acid mine drainage passive remediation system.

Science.gov (United States)

Johnson, D Barrie; Hallberg, Kevin B

2005-02-01

The compost bioreactor ("anaerobic cell") components of three composite passive remediation systems constructed to treat acid mine drainage (AMD) at the former Wheal Jane tin mine, Cornwall, UK were studied over a period of 16 months. While there was some amelioration of the preprocessed AMD in each of the three compost bioreactors, as evidenced by pH increase and decrease in metal concentrations, only one of the cells showed effective removal of the two dominant heavy metals (iron and zinc) present. With two of the compost bioreactors, concentrations of soluble (ferrous) iron draining the cells were significantly greater than those entering the reactors, indicating that there was net mobilisation (by reductive dissolution) of colloidal and/or solid-phase ferric iron compounds within the cells. Soluble sulfide was also detected in waters draining all three compost bioreactors which was rapidly oxidised, in contrast to ferrous iron. Oxidation and hydrolysis of iron, together with sulfide oxidation, resulted in reacidification of processed AMD downstream of the compost bioreactors in two of the passive treatment systems. The dominant cultivatable microorganism in waters draining the compost bioreactors was identified, via analysis of its 16S rRNA gene, as a Thiomonas sp. and was capable of accelerating the dissimilatory oxidation of both ferrous iron and reduced sulfur compounds. Sulfate-reducing bacteria (SRB) were also detected, although only in the bioreactor that was performing well were these present in significant numbers. This particular compost bioreactor had been shut down for 10 months prior to the monitoring period due to operational problems. This unforeseen event appears to have allowed more successful development of AMD-tolerant and other microbial populations with critical roles in AMD bioremediation, including neutrophilic SRB (nSRB), in this compost bioreactor than in the other two, where the throughput of AMD was not interrupted. This study has

Plant cell tissue culture: A potential source of chemicals

Energy Technology Data Exchange (ETDEWEB)

Scott, C.D.; Dougall, D.K.

1987-08-01

Higher plants produce many industrially important products. Among these are drugs and medicinal chemicals, essential oils and flavors, vegetable oils and fats, fine and specialty chemicals, and even some commodity chemicals. Although, currently, whole-plant extraction is the primary means of harvesting these materials, the advent of plant cell tissue culture could be a much more effective method of producing many types of phytochemicals. The use of immobilized plant cells in an advanced bioreactor configuration with excretion of the product into the reactor medium may represent the most straightforward way of commercializing such techniques for lower-value chemicals. Important research and development opportunities in this area include screening for plant cultures for nonmedical, lower-value chemicals; understanding and controlling plant cell physiology and biochemistry; optimizing effective immobilization methods; developing more efficient bioreactor concepts; and perfecting product extraction and purification techniques. 62 refs., 2 figs.

Brain Perfusion Changes in Intracerebral Hemorrhage

International Nuclear Information System (INIS)

Mititelu, R.; Mazilu, C.; Ghita, S.; Rimbu, A.; Marinescu, G.; Codorean, I.; Bajenaru, O.

2006-01-01

Full text: Purpose: Despite the latest advances in medical treatment and neuro critical care, patients suffering spontaneous intracerebral hemorrhage (SICH) still have a very poor prognosis, with a greater mortality and larger neurological deficits at the survivors than for ischemic stroke. Many authors have shown that there are many mechanisms involved in the pathology of SICH: edema, ischemia, inflammation, apoptosis. All of these factors are affecting brain tissue surrounding hematoma and are responsible of the progressive neurological deterioration; most of these damages are not revealed by anatomical imaging techniques. The aim of our study was to asses the role of brain perfusion SPECT in demonstrating perfusion changes in SICH patients. Method: 17 SICH pts were studied. All pts underwent same day CT and brain SPECT with 99mTcHMPAO, 24h-5d from onset of stroke. Results: 14/17 pts showed a larger perfusion defect than expected after CT. In 2 pts hematoma diameter was comparable on CT and SPECT; 1pt had quasinormal aspect of SPECT study. In pts with larger defects, SPECT revealed a large cold spot with similar size compared with CT, and a surrounding hypo perfused area. 6/17 pts revealed cortical hyper perfusion adjacent to hypo perfused area and corresponding to a normal-appearing brain tissue on CT. In 3 pts we found crossed cerebellar diaskisis.In 2 pts we found cortical hypo perfused area in the contralateral cortex, with normal appearing brain tissue on CT. Conclusions: Brain perfusion SPECT revealed different types of perfusion changes in the brain tissue surrounding hematoma. These areas contain viable brain tissue that may be a target for future ne uroprotective strategies. Further studies are definitely required to demonstrate prognostic significance of these changes, but we can conclude that brain perfusion SPECT can play an important role in SICH, by early demonstrating functional changes responsible of clinical deterioration, thus allowing prompt

An evaluation of different bioreactor configurations for continuous bio-ethanol production

International Nuclear Information System (INIS)

Ntihuga, Jean Nepomuscene; Senn, Thomas; Gschwind, Peter; Kohlus, Reinhard

2013-01-01

Highlights: • Two bioreactor configurations were constructed and compared. • Continuous bioethanol production was performed in both bioreactors. • Plate heat exchanger bioreactor was the best for solid mash fermentation. • Operational power costs of both bioreactors were different in small scale levels. • Further study needed for both bioreactors with optimized parameters. - Abstract: In this preliminary investigation, a so-called Blenke cascade and plate heat exchanger bioreactor configuration were compared in terms of mixing characteristics, contamination free process, operational power costs and overall performance. At room temperature, fermentation was initially started as batch run and switched to continuous operation, when the residual sugars within the reactor were detected to be C ⩽ 1% (g/L). Samples from both configurations were taken and analyzed for ethanol and residual sugar content, as well as for any infection of the fermentation and lactic acid content, respectively. Mixing characteristics were studied by the residence time distribution method. Both geometries behaved as a finite number n of continuous stirred tanks in series, behaving as a plug flow with superimposed axial dispersion. The number of tanks in series n obtained in the plate heat exchanger configuration was 1.5–3 times larger than those in the Blenke cascade. The average ethanol productivity was Q p = 3.07 (g/L h) and Q p = 2.31 (g/L h) for cascade and plate exchanger configuration, respectively. The analysis of operational power costs indicates relevant differences between the two reactors at laboratory scale; however, systems with different types of pumps and viscosities are compared. From an industrial scale point of view, specific operational costs decrease with scale-up, as no mechanical mixing is needed in the fermenters

Effect of aeration on the fermentative activity of Saccharomyces cerevisiae cultured in apple juice

OpenAIRE

Estela-Escalante, W.; Rychtera, M.; Melzoch, K.; Hatta-Sakoda, B.

2012-01-01

The influence of aeration on the fermentative activity of Saccharomyces cerevisiaeRTVE V 15-1-416 was studied in order to evaluate the synthesis of fermentation by-products. To achieve this, the strain was cultured in Erlenmeyer flasks and bioreactor containing sterilized and aroma removed apple juice. The chemical compounds produced during fermentations in shaken (200 min-¹) and static (without agitation) flasks and bioreactor, all in batch mode, were determined by GC and HPLC. The results s...

Developing a Benchmarking Process in Perfusion: A Report of the Perfusion Downunder Collaboration

Science.gov (United States)

Baker, Robert A.; Newland, Richard F.; Fenton, Carmel; McDonald, Michael; Willcox, Timothy W.; Merry, Alan F.

2012-01-01

Abstract: Improving and understanding clinical practice is an appropriate goal for the perfusion community. The Perfusion Downunder Collaboration has established a multi-center perfusion focused database aimed at achieving these goals through the development of quantitative quality indicators for clinical improvement through benchmarking. Data were collected using the Perfusion Downunder Collaboration database from procedures performed in eight Australian and New Zealand cardiac centers between March 2007 and February 2011. At the Perfusion Downunder Meeting in 2010, it was agreed by consensus, to report quality indicators (QI) for glucose level, arterial outlet temperature, and pCO2 management during cardiopulmonary bypass. The values chosen for each QI were: blood glucose ≥4 mmol/L and ≤10 mmol/L; arterial outlet temperature ≤37°C; and arterial blood gas pCO2 ≥ 35 and ≤45 mmHg. The QI data were used to derive benchmarks using the Achievable Benchmark of Care (ABC™) methodology to identify the incidence of QIs at the best performing centers. Five thousand four hundred and sixty-five procedures were evaluated to derive QI and benchmark data. The incidence of the blood glucose QI ranged from 37–96% of procedures, with a benchmark value of 90%. The arterial outlet temperature QI occurred in 16–98% of procedures with the benchmark of 94%; while the arterial pCO2 QI occurred in 21–91%, with the benchmark value of 80%. We have derived QIs and benchmark calculations for the management of several key aspects of cardiopulmonary bypass to provide a platform for improving the quality of perfusion practice. PMID:22730861

Non-contrast MRI perfusion angiosome in diabetic feet

Energy Technology Data Exchange (ETDEWEB)

Zheng, Jie [Cardiovascular Imaging Lab, Mallinckrodt Institute of Radiology, St. Louis, MO (United States); Hastings, Mary K.; Mueller, Michael J. [Washington University School of Medicine, The Program in Physical Therapy, St. Louis, MO (United States); Muccigross, David; Hildebolt, Charles F. [Washington University School of Medicine, Mallinckrodt Institute of Radiology, St. Louis, MO (United States); Fan, Zhaoyang [Cedars-Sinai Medical Center, Biomedical Imaging Research Institute, Los Angeles, CA (United States); Gao, Fabao [West China Hospital, Sichuan University, Department of Radiology, Chengdu (China); Curci, John [Washington University School of Medicine, The Department of Surgery, St. Louis, MO (United States)

2015-01-15

The purpose of this study is to develop a non-contrast magnetic resonance imaging (MRI) approach to evaluate skeletal muscle perfusion in the diabetic foot based on the concept of angiosomes of the foot. Five healthy volunteers and five participants with diabetes (HbA1c = 7.2 ± 1.8 %) without a history of peripheral artery disease were examined. The non-contrast perfusion measurements were performed during a toe flexion challenge. Absolute perfusion maps were created and two regions (medial and lateral) on the maps were segmented based on angiosomes. Regional difference in the perfusion of foot muscle was readily visualized in the MRI perfusion angiosomes during the challenge. In the participants with diabetes, the perfusion during toe flexion challenge was significantly lower than in healthy volunteers (P < 0.01). The average perfusion for the medial plantar region of the right foot was lower in subjects with diabetes (38 ± 9 ml/min/100 g) than in healthy subjects (93 ± 33 ml/min/100 g). Non-contrast MRI perfusion angiosome maps demonstrate the feasibility of determining regional perfusion in foot muscles during toe challenge and may facilitate evaluation of muscle perfusion in diabetic feet. (orig.)

Non-contrast MRI perfusion angiosome in diabetic feet

International Nuclear Information System (INIS)

Zheng, Jie; Hastings, Mary K.; Mueller, Michael J.; Muccigross, David; Hildebolt, Charles F.; Fan, Zhaoyang; Gao, Fabao; Curci, John

2015-01-01

The purpose of this study is to develop a non-contrast magnetic resonance imaging (MRI) approach to evaluate skeletal muscle perfusion in the diabetic foot based on the concept of angiosomes of the foot. Five healthy volunteers and five participants with diabetes (HbA1c = 7.2 ± 1.8 %) without a history of peripheral artery disease were examined. The non-contrast perfusion measurements were performed during a toe flexion challenge. Absolute perfusion maps were created and two regions (medial and lateral) on the maps were segmented based on angiosomes. Regional difference in the perfusion of foot muscle was readily visualized in the MRI perfusion angiosomes during the challenge. In the participants with diabetes, the perfusion during toe flexion challenge was significantly lower than in healthy volunteers (P < 0.01). The average perfusion for the medial plantar region of the right foot was lower in subjects with diabetes (38 ± 9 ml/min/100 g) than in healthy subjects (93 ± 33 ml/min/100 g). Non-contrast MRI perfusion angiosome maps demonstrate the feasibility of determining regional perfusion in foot muscles during toe challenge and may facilitate evaluation of muscle perfusion in diabetic feet. (orig.)

A Novel Pulsatile Bioreactor for Mechanical Stimulation of Tissue Engineered Cardiac Constructs

Directory of Open Access Journals (Sweden)

Günther Eissner

2011-07-01

Full Text Available After myocardial infarction, the implantation of stem cell seeded scaffolds on the ischemic zone represents a promising strategy for restoration of heart function. However, mechanical integrity and functionality of tissue engineered constructs need to be determined prior to implantation. Therefore, in this study a novel pulsatile bioreactor mimicking the myocardial contraction was developed to analyze the behavior of mesenchymal stem cells derived from umbilical cord tissue (UCMSC colonized on titanium-coated polytetrafluorethylene scaffolds to friction stress. The design of the bioreactor enables a simple handling and defined mechanical forces on three seeded scaffolds at physiological conditions. The compact system made of acrylic glass, Teflon®, silicone, and stainless steel allows the comparison of different media, cells and scaffolds. The bioreactor can be gas sterilized and actuated in a standard incubator. Macroscopic observations and pressure-measurements showed a uniformly sinusoidal pulsation, indicating that the bioreactor performed well. Preliminary experiments to determine the adherence rate and morphology of UCMSC after mechanical loadings showed an almost confluent cellular coating without damage on the cell surface. In summary, the bioreactor is an adequate tool for the mechanical stress of seeded scaffolds and offers dynamic stimuli for pre-conditioning of cardiac tissue engineered constructs in vitro.

Dual-energy perfusion-CT of pancreatic adenocarcinoma

International Nuclear Information System (INIS)

Klauß, M.; Stiller, W.; Pahn, G.; Fritz, F.; Kieser, M.; Werner, J.; Kauczor, H.U.; Grenacher, L.

2013-01-01

Purpose: To evaluate the feasibility of dual-energy CT (DECT)-perfusion of pancreatic carcinomas for assessing the differences in perfusion, permeability and blood volume of healthy pancreatic tissue and histopathologically confirmed solid pancreatic carcinoma. Materials and methods: 24 patients with histologically proven pancreatic carcinoma were examined prospectively with a 64-slice dual source CT using a dynamic sequence of 34 dual-energy (DE) acquisitions every 1.5 s (80 ml of iodinated contrast material, 370 mg/ml, flow rate 5 ml/s). 80 kV p , 140 kV p , and weighted average (linearly blended M0.3) 120 kV p -equivalent dual-energy perfusion image data sets were evaluated with a body-perfusion CT tool (Body-PCT, Siemens Medical Solutions, Erlangen, Germany) for estimating perfusion, permeability, and blood volume values. Color-coded parameter maps were generated. Results: In all 24 patients dual-energy CT-perfusion was. All carcinomas could be identified in the color-coded perfusion maps. Calculated perfusion, permeability and blood volume values were significantly lower in pancreatic carcinomas compared to healthy pancreatic tissue. Weighted average 120 kV p -equivalent perfusion-, permeability- and blood volume-values determined from DE image data were 0.27 ± 0.04 min −1 vs. 0.91 ± 0.04 min −1 (p −1 vs. 0.67 ± 0.05 *0.5 min −1 (p = 0.06) and 0.49 ± 0.07 min −1 vs. 1.28 ± 0.11 min −1 (p p the standard deviations of the kV p 120 kV p -equivalent values were manifestly smaller. Conclusion: Dual-energy CT-perfusion of the pancreas is feasible. The use of DECT improves the accuracy of CT-perfusion of the pancreas by fully exploiting the advantages of enhanced iodine contrast at 80 kV p in combination with the noise reduction at 140 kV p . Therefore using dual-energy perfusion data could improve the delineation of pancreatic carcinomas

Woodchip bioreactors effectively treat aquaculture effluent

Science.gov (United States)

Nutrients, in particular nitrogen and phosphorus, can create eutrophication problems in any watershed. Preventing water quality impairment requires controlling nutrients from both point-source and non-point source discharges. Woodchip bioreactors are one relatively new approach that can be utilized ...

In-vivo quantitative evaluation of perfusion zones and perfusion gradient in the deep inferior epigastric artery perforator flap

Science.gov (United States)

Saint-Cyr, Michel; Lakhiani, Chrisovalantis; Cheng, Angela; Mangum, Michael; Liang, Jinyang; Teotia, Sumeet; Livingston, Edward H.; Zuzak, Karel J.

2013-03-01

The selection of well-vascularized tissue during DIEP flap harvest remains controversial. While several studies have elucidated cross-midline perfusion, further characterization of perfusion to the ipsilateral hemiabdomen is necessary for minimizing rates of fat necrosis or partial fat necrosis in bilateral DIEP flaps. Eighteen patients (29 flaps) underwent DIEP flap harvest using a prospectively designed protocol. Perforators were marked and imaged with a novel system for quantitatively measuring tissue oxygenation, the Digital Light Hyperspectral Imager. Images were then analyzed to determine if perforator selection influenced ipsilateral flap perfusion. Flaps based on a single lateral row perforator (SLRP) were found to have a higher level of hemoglobin oxygenation in Zone I (mean %HbO2 = 76.1) compared to single medial row perforator (SMRP) flaps (%HbO2 = 71.6). Perfusion of Zone III relative to Zone I was similar between SLRP and SMRP flaps (97.4% vs. 97.9%, respectively). These differences were not statistically significant (p>0.05). Perfusion to the lateral edge of the flap was slightly greater for SLRP flaps compared SMRP flaps (92.1% vs. 89.5%, respectively). SMRP flaps had superior perfusion travelling inferiorly compared to SLRP flaps (88.8% vs. 83.9%, respectively). Overall, it was observed that flaps were better perfused in the lateral direction than inferiorly. Significant differences in perfusion gradients directed inferiorly or laterally were observed, and perforator selection influenced perfusion in the most distal or inferior aspects of the flap. This suggests broader clinical implications for flap design that merit further investigation.

Tissue engineering of cartilage using a mechanobioreactor exerting simultaneous mechanical shear and compression to simulate the rolling action of articular joints.

Science.gov (United States)

Shahin, Kifah; Doran, Pauline M

2012-04-01

The effect of dynamic mechanical shear and compression on the synthesis of human tissue-engineered cartilage was investigated using a mechanobioreactor capable of simulating the rolling action of articular joints in a mixed fluid environment. Human chondrocytes seeded into polyglycolic acid (PGA) mesh or PGA-alginate scaffolds were precultured in shaking T-flasks or recirculation perfusion bioreactors for 2.5 or 4 weeks prior to mechanical stimulation in the mechanobioreactor. Constructs were subjected to intermittent unconfined shear and compressive loading at a frequency of 0.05 Hz using a peak-to-peak compressive strain amplitude of 2.2% superimposed on a static axial compressive strain of 6.5%. The mechanical treatment was carried out for up to 2.5 weeks using a loading regime of 10 min duration each day with the direction of the shear forces reversed after 5 min and release of all loading at the end of the daily treatment period. Compared with shaking T-flasks and mechanobioreactor control cultures without loading, mechanical treatment improved the amount and quality of cartilage produced. On a per cell basis, synthesis of both major structural components of cartilage, glycosaminoglycan (GAG) and collagen type II, was enhanced substantially by up to 5.3- and 10-fold, respectively, depending on the scaffold type and seeding cell density. Levels of collagen type II as a percentage of total collagen were also increased after mechanical treatment by up to 3.4-fold in PGA constructs. Mechanical treatment had a less pronounced effect on the composition of constructs precultured in perfusion bioreactors compared with perfusion culture controls. This work demonstrates that the quality of tissue-engineered cartilage can be enhanced significantly by application of simultaneous dynamic mechanical shear and compression, with the greatest benefits evident for synthesis of collagen type II. Copyright © 2011 Wiley Periodicals, Inc.

Perfusion vector - a new method to quantify myocardial perfusion scintigraphy images: a simulation study with validation in patients

DEFF Research Database (Denmark)

Minarik, David; Senneby, Martin; Wollmer, Per

2015-01-01

Background The interpretation of myocardial perfusion scintigraphy (MPS) largely relies on visual assessment by the physician of the localization and extent of a perfusion defect. The aim of this study was to introduce the concept of the perfusion vector as a new objective quantitative method...

Bacterial community dynamics in a rumen fluid bioreactor during in-vitro cultivation.

Science.gov (United States)

Zapletalová, Martina; Kašparovská, Jitka; Křížová, Ludmila; Kašparovský, Tomáš; Šerý, Omar; Lochman, Jan

2016-09-20

To study the various processes in the rumen the in vitro techniques are widely used to realize more controlled and reproducible conditions compared to in vivo experiments. Mostly, only the parameters like pH changes, volatile fatty acids content or metabolite production are monitored. In this study we examine the bacterial community dynamics of rumen fluid in course of ten day cultivation realize under standard conditions described in the literature. Whereas the pH values, total VFA content and A/P ratio in bioreactor were consistent with natural conditions in the rumen, the mean redox-potential values of -251 and -243mV were much more negative. For culture-independent assessment of bacterial community composition, the Illumina MiSeq results indicated that the community contained 292 bacterial genera. In course of ten days cultivation a significant changes in the microbial community were measured when Bacteroidetes to Firmicutes ratio changed from 3.2 to 1.2 and phyla Proteobacteria and Actinobacteria represented by genus Bifidobacterium and Olsenella significantly increased. The main responsible factor of these changes seems to be very low redox potential in bioreactor together with accumulation of simple carbohydrates in milieu as a result of limited excretion of fermented feed and absence of nutrient absorbing mechanisms. Copyright © 2016 Elsevier B.V. All rights reserved.

Placental perfusion - a human alternative

DEFF Research Database (Denmark)

Mose, Tina; Knudsen, Lisbeth E

2006-01-01

Foetal exposures to environmental and medicinal products have impact on the growth of the foetus (e.g. cigarette smoke) and development of organs (e.g. methylmercury and Thalidomide). Perfusion studies of the human term placenta enable investigation of placental transport of chemical substances...... between the mother and foetus. Dual perfusion of a single cotyledon in the human placenta can contribute to a better understanding of the placental barrier, transport rate and mechanisms of different substances and placental metabolism. The perfusion system has recently been established in Copenhagen...

The Groningen hypothermic liver perfusion pump : Functional evaluation of a new machine perfusion system

NARCIS (Netherlands)

van der Plaats, A.; Maathuis, M. H. J.; Hart, N. A. 't; Bellekom, A. A.; Hofker, H. S.; van der Houwen, E. B.; Verkerke, G. J.; Leuvenink, H. G. D.; Verdonck, P.; Ploeg, R. J.; Rakhorst, G.

2006-01-01

To improve preservation of donor livers, we have developed a portable hypothermic machine perfusion (HMP) system as an alternative for static cold storage. A prototype of the system was built and evaluated on functionality. Evaluation criteria included 24 h of adequate pressure controlled perfusion,

Evaluation of Productivity of Zymotis Solid-State Bioreactor Based on Total Reactor Volume

Directory of Open Access Journals (Sweden)

Oscar F. von Meien

2002-01-01

Full Text Available In this work a method of analyzing the performance of solid-state fermentation bioreactors is described. The method is used to investigate the optimal value for the spacing between the cooling plates of the Zymotis bioreactor, using simulated fermentation data supplied by a mathematical model. The Zymotis bioreactor has good potential for those solid-state fermentation processes in which the substrate bed must remain static. The current work addresses two design parameters introduced by the presence of the internal heat transfer plates: the width of the heat transfer plate, which is governed by the amount of heat to be removed and the pressure drop of the cooling water, and the spacing between these heat transfer plates. In order to analyze the performance of the bioreactor a productivity term is introduced that takes into account the volume occupied within the bioreactor by the heat transfer plates. As part of this analysis, it is shown that, for logistic growth kinetics, the time at which the biomass reaches 90 % of its maximum possible value is a good estimate of the optimum harvesting time for maximizing productivity. Application of the productivity analysis to the simulated fermentation results suggests that, with typical fast growing fungi ( = 0.324 h–1, the optimal spacing between heat transfer plates is of the order of 6 cm. The general applicability of this approach to evaluate the productivity of solid-state bioreactors is demonstrated.

Increased perfusion pressure enhances the expression of endothelin (ETB) and angiotensin II (AT1, AT2) receptors in rat mesenteric artery smooth muscle cells

DEFF Research Database (Denmark)

Lindstedt, Isak; Xu, Cang-Bao; Zhang, Yaping

2009-01-01

and luminally perfused in a perfusion chamber. After either exposure to no ("organ culture" (0 mmHg)), normal (85/75 mmHg) or high pressure (160/150 mmHg) at constant flow for 1-17 h, the vessel segments were snap frozen and real-time polymerase chain reaction was performed to quantify the ET- and AT-receptor m...

Ultra-micro aqua bioreactor systems for modifying edible oils and fats; Shokuyo yushi kaishitsuyo chobisuikei bioreactor

Energy Technology Data Exchange (ETDEWEB)

Kurashige, J. [Ajinomoto Co. Inc., Tokyo (Japan)

1995-10-20

Practical solvent-free bioreactor systems using immobilized lipases have been constructed to convert palm oil to high quality foodstuff oil without quality deterioration through hydrolysis of triglycerides in oil. To avoid hydrolysis, moisture level of substrate oil has to be maintained at less than the solubility level of water in oil, which we call ultra-micro aqueous level. On the other hand, lipase is well known to manifest its activities mostly at the interface between oil and water phases. To make lipase manifest its activities at the ultra-micro aqueous oil phase, the novel bioreactor systems with the new immobilizing method of lipase together with activator on-to hydrophylic carriers, and without a drying procedure have been developed. These biochemical accomplishments show high promises for efficient convention of edible fats and oils to highly valuable foodstuff, which can not be attained by means of chemical or physical methods. 29 refs., 9 figs., 4 tabs.

Vicarious audiovisual learning in perfusion education.

Science.gov (United States)

Rath, Thomas E; Holt, David W

2010-12-01

Perfusion technology is a mechanical and visual science traditionally taught with didactic instruction combined with clinical experience. It is difficult to provide perfusion students the opportunity to experience difficult clinical situations, set up complex perfusion equipment, or observe corrective measures taken during catastrophic events because of patient safety concerns. Although high fidelity simulators offer exciting opportunities for future perfusion training, we explore the use of a less costly low fidelity form of simulation instruction, vicarious audiovisual learning. Two low fidelity modes of instruction; description with text and a vicarious, first person audiovisual production depicting the same content were compared. Students (n = 37) sampled from five North American perfusion schools were prospectively randomized to one of two online learning modules, text or video.These modules described the setup and operation of the MAQUET ROTAFLOW stand-alone centrifugal console and pump. Using a 10 question multiple-choice test, students were assessed immediately after viewing the module (test #1) and then again 2 weeks later (test #2) to determine cognition and recall of the module content. In addition, students completed a questionnaire assessing the learning preferences of today's perfusion student. Mean test scores from test #1 for video learners (n = 18) were significantly higher (88.89%) than for text learners (n = 19) (74.74%), (p audiovisual learning modules may be an efficacious, low cost means of delivering perfusion training on subjects such as equipment setup and operation. Video learning appears to improve cognition and retention of learned content and may play an important role in how we teach perfusion in the future, as simulation technology becomes more prevalent.

Bioreactors for fixation and effective utilization of carbon dioxide gas. Tansan gas no koteiter dot yuko riyo no tame no bio reactor

Energy Technology Data Exchange (ETDEWEB)

Miyamoto, K. (Osaka University, Osaka (Japan). Faculty of Pharmaceutical Science); Benemann, J. (California University, CA (USA))

1991-06-01

As for a preventive countermeasure against the global warming, experiments and studies have been conducted on the bioreactors to fix carbon dioxide gas recovered from the concentric and large scale generating sources such as thermal power plamts in a form of carbohydrate by means of the culture of microbial algae. By using the Vertical Tube Reactors (VTR) culturing apparatus, a variety of microbial algae were cultivated and experiments were performed on the relationship of biomass productivity and absorption rate of carbon dioxide gas indoors and outdoors. Consequently, it was found that when the flow rate of carbon dioxide gas is adjusted to make the biomass productivity of filament type Nostoc maximum,the inlet and outlet concentrations of carbon dioxide gas were 0.7% and 0.05% respectively with the absorption rate of more than 90%. From the standpoint of fixation and effective utilization of carbon dioxide gas, the above rate of removal is one of the important parameters and it will be necessary in future to compare the rates of removal of carbon dioxide gas among various types of bioreactors as a function of operating condition. 9 refs., 6 figs., 2 tabs.

BIOREACTOR WITH LID FOR EASY ACCESS TO INCUBATION CAVITY

DEFF Research Database (Denmark)

2012-01-01

There is provided a bioreactor which is provided with a lid (13) that facilitates access to the incubation cavity. Specifically the end wall of the incubation cavity is constituted by the lid (13) so that removal of the cap renders the incubation cavity fully accessible.......There is provided a bioreactor which is provided with a lid (13) that facilitates access to the incubation cavity. Specifically the end wall of the incubation cavity is constituted by the lid (13) so that removal of the cap renders the incubation cavity fully accessible....

Pulmonary ventilation and perfusion abnormalities and ventilation perfusion imbalance in children with pulmonary atresia or extreme tetralogy of Fallot

Energy Technology Data Exchange (ETDEWEB)

Dowdle, S.C.; Human, D.G.; Mann, M.D. (Univ. of Cape Town (South Africa))

1990-08-01

Xenon-133 lung ventilation and perfusion scans were done preoperatively after cardiac catheterization and cineangiocardiography in 19 children; 6 had pulmonary atresia with an intact ventricular septum and hypoplastic right ventricle, 4 pulmonary atresia with associated complex univentricular heart, and 9 extreme Tetralogy of Fallot. The four patients with discrepancies in the sizes of the left and right pulmonary arteries on angiography had marked asymmetry of pulmonary perfusion and ventilation-perfusion imbalance on scintigraphy. Similar degrees of asymmetry and imbalance were present in 6 of the 15 children with equal-size pulmonary vessels. Asymmetry of pulmonary perfusion and ventilation-perfusion imbalance were associated with a poor prognosis.

Pulmonary ventilation and perfusion abnormalities and ventilation perfusion imbalance in children with pulmonary atresia or extreme tetralogy of Fallot

International Nuclear Information System (INIS)

Dowdle, S.C.; Human, D.G.; Mann, M.D.

1990-01-01

Xenon-133 lung ventilation and perfusion scans were done preoperatively after cardiac catheterization and cineangiocardiography in 19 children; 6 had pulmonary atresia with an intact ventricular septum and hypoplastic right ventricle, 4 pulmonary atresia with associated complex univentricular heart, and 9 extreme Tetralogy of Fallot. The four patients with discrepancies in the sizes of the left and right pulmonary arteries on angiography had marked asymmetry of pulmonary perfusion and ventilation-perfusion imbalance on scintigraphy. Similar degrees of asymmetry and imbalance were present in 6 of the 15 children with equal-size pulmonary vessels. Asymmetry of pulmonary perfusion and ventilation-perfusion imbalance were associated with a poor prognosis

Predictions for optimal mitigation of paracrine inhibitory signalling in haemopoietic stem cell cultures.

Science.gov (United States)

Berry, Joseph D; Godara, Pankaj; Liovic, Petar; Haylock, David N

2015-04-16

Recent studies in the literature have highlighted the critical role played by cell signalling in determining haemopoietic stem cell (HSC) fate within ex vivo culture systems. Stimulatory signals can enhance proliferation and promote differentiation, whilst inhibitory signals can significantly limit culture output. Numerical models of various mitigation strategies are presented and applied to determine effectiveness of these strategies toward mitigation of paracrine inhibitory signalling inherent in these culture systems. The strategies assessed include mixing, media-exchange, fed-batch and perfusion. The models predict that significant spatial concentration gradients exist in typical cell cultures, with important consequences for subsequent cell expansion. Media exchange is shown to be the most effective mitigation strategy, but remains labour intensive and difficult to scale-up for large culture systems. The fed-batch strategy is only effective at very small Peclet number, and its effect is diminished as the cell culture volume grows. Conversely, mixing is effective at high Peclet number, and ineffective at low Peclet number. The models predict that cell expansion in fed-batch cultures becomes independent of increasing dilution rate, consistent with experimental results previously reported in the literature. In contrast, the models predict that increasing the flow rate in perfused cultures will lead to increased cell expansion, indicating the suitability of perfusion for use as an automated, tunable strategy. The effect of initial cell seeding density is also investigated, with the model showing that perfusion outperforms dilution for all densities considered. The models predict that the impact of inhibitory signalling in HSC cultures can be mitigated against using media manipulation strategies, with the optimal strategy dependent upon the protein diffusion time-scale relative to the media manipulation time-scale. The key messages from this study can be applied to

Efficacy of Bioremediation of Agricultural Runoff Using Bacterial Communities in Woodchip Bioreactors.

Science.gov (United States)

Mortensen, Z. H.; Leandro, M.; Silveus, J. M.

2016-12-01

California's agricultural sector is fundamental in the State's economic growth and is responsible for supplying a large portion of the country's produce. In order to meet the market's demand for crop production the region's agrarian landscape requires an abundance of nutrient rich irrigation. The resultant agricultural effluent is a source of increased nutrient content in California's watershed and groundwater systems, promoting eutrophication and contributing to negative impacts on local ecosystems and human health. Previous studies have examined the denitrification potential of woodchip bioreactors. However, research has been deficient regarding specific variables that may affect the remediation process. To evaluate the efficacy of woodchip bioreactors in remediating waters containing high nitrate concentrations, denitrification rates were examined and parameters such as temperature, laminar flow, and hydraulic residence times were measured to identify potential methods for increasing denitrification efficiency. By measuring the rate of denitrification in a controlled environment where potentially confounding factors can be manipulated, physical components affecting the efficiency of woodchip bioreactors were examined to assess effects. Our research suggests the implementation of woodchip bioreactors to treat agricultural runoff would significantly reduce the concentration of nitrate in agricultural effluent and contribute to the mitigation of negative impacts associated with agricultural irrigation. Future research should focus on the ability of woodchip bioreactors to successfully remediate other agricultural pollutants, such as phosphates and pesticides, to optimize the efficiency of the bioremediation process.

Modeling of hydrodynamics in hollow fiber membrane bioreactor for mammalian cells cultivation

Directory of Open Access Journals (Sweden)

N. V. Menshutina

2016-01-01

Full Text Available The mathematical modelling in CFD-packages are powerfull instrument for design and calculation of any engineering tasks. CFD-package contains the set of programs that allow to model the different objects behavior based on the mathematical lows. ANSYS Fluent are widely used for modelling of biotechnological and chemical-technological processes. This package is convenient to describe their hydrodynamics. As cell cultivation is one of the actual scientific direction in modern biotechnology ANSYS Fluent was used to create the model of hollow fiber membrane bioreactor. The fibers are hollow cylindrical membrane to be used for cell cultivation. The criterion of process effectiveness for cell growth is full filling of the membrane surface by cells in the bioreactor. While the cell growth the fiber permeability is decreased which effects to feed flow through membrane pores. The specific feature of this process is to ensure such feed flow to deliver the optimal nutrition for the cells on the external membrane surface. The velocity distribution inside the fiber and in all bioreactor as a whole has been calculated based on mass an impulse conservation equations taking into account the mathematical model assumptions. The hydrodynamics analysis in hollow fiber membrane bioreactor is described by the three-dimensional model created in ANSYS Fluent. The specific features of one membrane model are considered and for whole bioreactor too.

Combination of Electrochemical Processes with Membrane Bioreactors for Wastewater Treatment and Fouling Control: A Review

OpenAIRE

Ensano, Benny M. B.; Borea, Laura; Naddeo, Vincenzo; Belgiorno, Vincenzo; de Luna, Mark D. G.; Ballesteros, Florencio C.

2016-01-01

This paper provides a critical review about the integration of electrochemical processes into membrane bioreactors (MBR) in order to understand the influence of these processes on wastewater treatment performance and membrane fouling control. The integration can be realized either in an internal or an external configuration. Electrically enhanced membrane bioreactors or electro membrane bioreactors (eMBRs) combine biodegradation, electrochemical and membrane filtration processes into one syst...

Combination of electrochemical processes with membrane bioreactors for wastewater treatment and fouling control: A review

OpenAIRE

Benny Marie B. Ensano; Laura Borea; Vincenzo Naddeo; Vincenzo Belgiorno; Mark Daniel G. de Luna; Mark Daniel G. de Luna; Florencio C. Ballesteros, Jr.; Florencio C. Ballesteros, Jr.

2016-01-01

This paper provides a critical review about the integration of electrochemical processes into membrane bioreactors (MBR) in order to understand the influence of these processes on wastewater treatment performance and membrane fouling control. The integration can be realized either in an internal or an external configuration. Electrically enhanced membrane bioreactors or electro membrane bioreactors (eMBRs) combine biodegradation, electrochemical and membrane filtration processes into one syst...

Sulfate-reducing bacteria in anaerobic bioreactors

NARCIS (Netherlands)

Oude Elferink, S.J.W.H.

1998-01-01

The treatment of industrial wastewaters containing high amounts of easily degradable organic compounds in anaerobic bioreactors is a well-established process. Similarly, wastewaters which in addition to organic compounds also contain sulfate can be treated in this way. For a long time, the

LANDFILL BIOREACTOR PERFORMANCE, SECOND INTERIM REPORT

Science.gov (United States)

A bioreactor landfill is a landfill that is operated in a manner that is expected to increase the rate and extent of waste decomposition, gas generation, and settlement compared to a traditional landfill. This Second Interim Report was prepared to provide an interpretation of fie...

Clinical application of cerebral dynamic perfusion studies

International Nuclear Information System (INIS)

DeLand, F.H.

1975-01-01

Radionuclide cerebral perfusion studies are assuming a far greater importance in the detection and differential diagnosis of cerebral lesions. Perfusion studies not only contribute to the differential diagnosis of lesions but in certain cases are the preferred methods by which more accurate clinical interpretations can be made. The characteristic blood flow of arterio-venous malformations readily differentiates this lesion from neoplasms. The decreased perfusion or absent perfusion observed in cerebral infarctions is diagnostic without concurrent evidence from static images. Changes in rates and direction of blood flow contribute fundamental information to the status of stenosis and vascular occlusion and, in addition, offer valuable information on the competency and routes of collateral circulation. The degree of cerebral perfusion after cerebral vascular accidents appears to be directly related to patient recovery, particularly muscular function. Cerebral perfusion adds a new parameter in the diagnosis of subdural haematomas and concussion and in the differentiation of obscuring radioactivity from superficial trauma. Although pictorial displays of perfusion blood flow will offer information in most cerebral vascular problems, the addition of computer analysis better defines temporal relationships of regional blood flow, quantitative changes in flow and the detection of the more subtle increases or decreases in cerebral blood flow. The status of radionuclide cerebral perfusion studies has taken on an importance making it the primary modality for the diagnosis of cerebral lesions. (author)

Lactic acid Production with in situ Extraction in Membrane Bioreactor

Directory of Open Access Journals (Sweden)

Hamidreza Ghafouri Taleghani

2017-01-01

Full Text Available Background and Objective: Lactic acid is widely used in the food, chemical and pharmaceutical industries. The major problems associated with lactic acid production are substrate and end-product inhibition, and by-product formation. Membrane technologyrepresents one of the most effective processes for lactic acid production. The aim of this work is to increase cell density and lactic acid productivity due to reduced inhibition effect of substrate and product in membrane bioreactor.Material and Methods: In this work, lactic acid was produced from lactose in membrane bioreactor. A laboratory scale membrane bioreactor was designed and fabricated. Five types of commercial membranes were tested at the same operating conditions (transmembrane pressure: 500 KPa and temperature: 25°C. The effects of initial lactose concentration and dilution rate on biomass growth, lactic acid production and substrate utilization were evaluated.Results and Conclusion: The high lactose retention of 79% v v-1 and low lactic acid retention of 22% v v-1 were obtained with NF1 membrane; therefore, this membrane was selected for membrane bioreactor. The maximal productivity of 17.1 g l-1 h-1 was obtainedwith the lactic acid concentration of 71.5 g l-1 at the dilution rate of 0.24 h−1. The maximum concentration of lactic acid was obtained at the dilution rate of 0.04 h−1. The inhibiting effect of lactic acid was not observed at high initial lactose concentration. The critical lactose concentration at which the cell growth severely hampered was 150 g l-1. This study proved that membrane bioreactor had great advantages such as elimination of substrate and product inhibition, high concentration of process substrate, high cell density,and high lactic acid productivity.Conflict of interest: There is no conflict of interest.

Effect of dynamic three-dimensional culture on osteogenic potential of human periodontal ligament-derived mesenchymal stem cells entrapped in alginate microbeads.

Science.gov (United States)

Vecchiatini, R; Penolazzi, L; Lambertini, E; Angelozzi, M; Morganti, C; Mazzitelli, S; Trombelli, L; Nastruzzi, C; Piva, R

2015-08-01

Bioreactors are devices that efficiently create an environment that enables cell cultures to grow in a three-dimensional (3D) context mimicking in vivo conditions. In this study, we investigate the effect of dynamic fluid flow on the osteogenic potential of human mesenchymal stem cells obtained from periodontal ligament and entrapped in alginate microbeads. After proper immunophenotyping, cells were encapsulated in barium alginate, cultured in 3D static or 3D dynamic conditions represented by a bioreactor system. Calcein-AM/propidium iodide staining was used to assess cellular viability. Quantitative real-time polymerase chain reaction was used to analyze the expression of osteogenic markers (Runx2 and COL1). Alizarin Red S staining and the Fourier transform infrared spectroscopy were used to assess mineral matrix deposition. Optimal encapsulation procedure, in terms of polymer pumping rate, distance from droplet generator to the gelling bath and atomizing airflow was assessed. Cell viability was not affected by encapsulation in alginate microbeads. Bioreactor cell exposure was effective in anticipating osteogenic differentiation and improving mineral matrix deposition. For the first time human mesenchymal stem cells obtained from periodontal ligaments encapsulated in alginate microbeads were cultured in a bioreactor system. This combination could represent a promising strategy to create a cell-based smart system with enhanced osteogenic potential useful for many different dental applications. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Hydrodynamic performance of a single-use aerated stirred bioreactor in animal cell culture: applications of tomography, dynamic gas disengagement (DGD), and CFD.

Science.gov (United States)

Kazemzadeh, Argang; Elias, Cynthia; Tamer, Melih; Ein-Mozaffari, Farhad

2018-05-01

The hydrodynamics of gas-liquid two-phase flow in a single-use bioreactor were investigated in detail both experimentally and numerically. Electrical resistance tomography (ERT) and dynamic gas disengagement (DGD) combined with computational fluid dynamics (CFD) were employed to assess the effect of the volumetric gas flow rate and impeller speed on the gas-liquid flow field, local and global gas holdup values, and Sauter mean bubble diameter. From the results obtained from DGD coupled with ERT, the bubble sizes were determined. The experimental data indicated that the total gas holdup values increased with increasing both the rotational speed of impeller and volumetric gas flow rate. Moreover, the analysis of the flow field generated inside the aerated stirred bioreactor was conducted using CFD results. Overall, a more uniform distribution of the gas holdup was obtained at impeller speeds ≥ 100 rpm for volumetric gas flow rates ≥ 1.6 × 10 -5  m 3 /s.

Cyclic uniaxial compression of human stem cells seeded on a bone biomimetic nanocomposite decreases anti-osteogenic commitment evoked by shear stress.

Science.gov (United States)

Baumgartner, Walter; Schneider, Isabelle; Hess, Samuel C; Stark, Wendelin J; Märsmann, Sonja; Brunelli, Marzia; Calcagni, Maurizio; Cinelli, Paolo; Buschmann, Johanna

2018-04-05

Chemical supplementation of culture media to induce differentiation of adult stem cells seeded on a scaffold may mask other differentiation triggers such as scaffold stiffness, chemical composition or mechanical stimulation. However, stem cells can be differentiated towards osteoblasts without any supplementation given an appropriate osteogenic scaffold and an adequate mechanical stimulation. Electrospun meshes of poly-lactic-co-glycolic acid and amorphous calcium phosphate nanoparticles (PLGA/aCaP) in a weight ratio of 60:40 were seeded with human adipose-derived stem cells (ASCs) and cultured in DMEM. After two weeks of static cultivation, they were either further cultivated statically for another two weeks (group 1), or placed in a Bose® bioreactor with a flow rate per area of 0.16 mL cm -2 min 1 (group 2). Furthermore, group 3 was also cultivated under perfusion, however, with an additional uniaxial cyclic compression. Stiffness of the scaffolds was assessed as a function of time. After a total of four weeks, minimum stem cell criteria markers as well as typical markers for osteogenesis, endothelial cell differentiation, adipogenesis and chondrogenesis were analyzed by quantitative real-time PCR, cell distribution within the scaffolds by histology and protein expression by immunohistochemistry. Dynamic conditions (perfusion ± uniaxial cyclic compression) significantly upregulated gene and protein expression of PPAR-γ-2 compared to static cultivation, while osteogenic markers were slightly downregulated. However, the compression in the perfusion bioreactor favored osteogenesis compared to mere perfusion as indicated by upregulation of ALP, Runx2 and collagen I. This behavior was not only attributed to the compressive load, but also to the significant increase in stiffness of the scaffold. Furthermore, CD105 was significantly upregulated under compression. Although an osteogenic electrospun composite material with an organic (PLGA) and an inorganic phase

A symbiotic gas exchange between bioreactors enhances microalgal biomass and lipid productivities: taking advantage of complementary nutritional modes.

Science.gov (United States)

Santos, C A; Ferreira, M E; da Silva, T Lopes; Gouveia, L; Novais, J M; Reis, A

2011-08-01

This paper describes the association of two bioreactors: one photoautotrophic and the other heterotrophic, connected by the gas phase and allowing an exchange of O(2) and CO(2) gases between them, benefiting from a symbiotic effect. The association of two bioreactors was proposed with the aim of improving the microalgae oil productivity for biodiesel production. The outlet gas flow from the autotrophic (O(2) enriched) bioreactor was used as the inlet gas flow for the heterotrophic bioreactor. In parallel, the outlet gas flow from another heterotrophic (CO(2) enriched) bioreactor was used as the inlet gas flow for the autotrophic bioreactor. Aside from using the air supplied from the auto- and hetero-trophic bioreactors as controls, one mixotrophic bioreactor was also studied and used as a model, for its claimed advantage of CO(2) and organic carbon being simultaneously assimilated. The microalga Chlorella protothecoides was chosen as a model due to its ability to grow under different nutritional modes (auto, hetero, and mixotrophic), and its ability to attain a high biomass productivity and lipid content, suitable for biodiesel production. The comparison between heterotrophic, autotrophic, and mixotrophic Chlorella protothecoides growth for lipid production revealed that heterotrophic growth achieved the highest biomass productivity and lipid content (>22%), and furthermore showed that these lipids had the most suitable fatty acid profile in order to produce high quality biodiesel. Both associations showed a higher biomass productivity (10-20%), when comparing the two separately operated bioreactors (controls) which occurred on the fourth day. A more remarkable result would have been seen if in actuality the two bioreactors had been inter-connected in a closed loop. The biomass productivity gain would have been 30% and the lipid productivity gain would have been 100%, as seen by comparing the productivities of the symbiotic assemblage with the sum of the two

Analysis of the efficiency of recombinant Escherichia coli strain cultivation in a gas-vortex bioreactor.

Science.gov (United States)

Savelyeva, Anna V; Nemudraya, Anna A; Podgornyi, Vladimir F; Laburkina, Nadezhda V; Ramazanov, Yuriy A; Repkov, Andrey P; Kuligina, Elena V; Richter, Vladimir A

2017-09-01

The levels of aeration and mass transfer are critical parameters required for an efficient aerobic bioprocess, and directly depend on the design features of exploited bioreactors. A novel apparatus, using gas vortex for aeration and mass transfer processes, was constructed in the Center of Vortex Technologies (Novosibirsk, Russia). In this paper, we compared the efficiency of recombinant Escherichia coli strain cultivation using novel gas-vortex technology with conventional bioprocess technologies such as shake flasks and bioreactors with mechanical stirrers. We demonstrated that the system of aeration and agitation used in gas-vortex bioreactors provides 3.6 times higher volumetric oxygen transfer coefficient in comparison with mechanical bioreactor. The use of gas-vortex bioreactor for recombinant E. coli strain cultivation allows to increase the efficiency of target protein expression at 2.2 times for BL21(DE3)/pFK2 strain and at 3.5 times for auxotrophic C600/pRT strain (in comparison with stirred bioreactor). © 2016 International Union of Biochemistry and Molecular Biology, Inc.

Dynamic perfusion CT: Optimizing the temporal resolution for the calculation of perfusion CT parameters in stroke patients

Energy Technology Data Exchange (ETDEWEB)

Kaemena, Andreas [Department of Radiology, Charite-Medical University Berlin, Augustenburger Platz 1, D-13353 Berlin (Germany)], E-mail: andreas.kaemena@charite.de; Streitparth, Florian; Grieser, Christian; Lehmkuhl, Lukas [Department of Radiology, Charite-Medical University Berlin, Augustenburger Platz 1, D-13353 Berlin (Germany); Jamil, Basil [Department of Radiotherapy, Charite-Medical University Berlin, Schumannstr. 20/21, D-10117 Berlin (Germany); Wojtal, Katarzyna; Ricke, Jens; Pech, Maciej [Department of Radiology, Charite-Medical University Berlin, Augustenburger Platz 1, D-13353 Berlin (Germany)

2007-10-15

Purpose: To assess the influence of different temporal sampling rates on the accuracy of the results from cerebral perfusion CTs in patients with an acute ischemic stroke. Material and methods: Thirty consecutive patients with acute stroke symptoms received a dynamic perfusion CT (LightSpeed 16, GE). Forty millilitres of iomeprol (Imeron 400) were administered at an injection rate of 4 ml/s. After a scan delay of 7 s, two adjacent 10 mm slices at 80 kV and 190 mA were acquired in a cine mode technique with a cine duration of 49 s. Parametric maps for the blood flow (BF), blood volume (BV) and mean transit time (MTT) were calculated for temporal sampling intervals of 0.5, 1, 2, 3 and 4 s using GE's Perfusion 3 software package. In addition to the quantitative ROI data analysis, a visual perfusion map analysis was performed. Results: The perfusion analysis proved to be technically feasible with all patients. The calculated perfusion values revealed significant differences with regard to the BF, BV and MTT, depending on the employed temporal resolution. The perfusion contrast between ischemic lesions and healthy brain tissue decreased continuously at the lower temporal resolutions. The visual analysis revealed that ischemic lesions were best depicted with sampling intervals of 0.5 and 1 s. Conclusion: We recommend a temporal scan resolution of two images per second for the best detection and depiction of ischemic areas.

Comparison between moving bed-membrane bioreactor (MB-MBR) and membrane bioreactor (MBR) systems: influence of wastewater salinity variation.

Science.gov (United States)

Di Trapani, Daniele; Di Bella, Gaetano; Mannina, Giorgio; Torregrossa, Michele; Viviani, Gaspare

2014-06-01

Two pilot plant systems were investigated for the treatment of wastewater subject to a gradual increase of salinity. In particular, a membrane bioreactor (MBR) and a moving bed biofilm membrane bioreactor (MB-MBR) were analyzed. Carbon and ammonium removal, kinetic constants and membranes fouling rates have been assessed. Both plants showed very high efficiency in terms of carbon and ammonium removal and the gradual salinity increase led to a good acclimation of the biomass, as confirmed by the respirometric tests. Significant biofilm detachments from carriers were experienced, which contributed to increase the irreversible superficial cake deposition. However, this aspect prevented the pore fouling tendency in the membrane module of MB-MBR system. On the contrary, the MBR pilot, even showing a lower irreversible cake deposition, was characterized by a higher pore fouling tendency. Copyright © 2014 Elsevier Ltd. All rights reserved.

Membrane bioreactors for enzymatic hydrolysis of lactose; Idrolisi enzimatica del lattosio con bioreattori a membrana

Energy Technology Data Exchange (ETDEWEB)

Pizzichini, M; Pilloton, R [ENEA, Casaccia (Italy). Area Energia e Innovazione; Pontecorvo, M; Mignogna, G; Fortunato, A; Beone, F

1993-03-01

Bioreactor systems obtained by cell or enzyme immobilization offer many advantages compared with native enzyme, intact cell systems or other biocatalysts. Thus, many attempts have been made to design and use new types of bioreactor systems in order to improve performance, enhance productivity and reduce environmental impacts. Membrane bioreactors, obtained by physical immobilization of biocatalysts, in polymeric membrane support, offer such practical advantages as: a continuous separation and transformation process with low product inhibition and suitable hydraulic configuration (backflushing recycling, ultrafiltrating). Specific membrane modules (Amicon VitaFiber), for bioreactor applications are being commercialized. Beta-galctosidase enzyme has successfully been immobilized in a hollow fiber and in ceramic modules to hydrolyze lactose in waste whey. This technical report presents the general properties and performances (permeability, washing procedures, hydraulic configurations, physical and chemical properties) of both, polymeric and ceramic supports, enzyme kinetics, physical and covalent immobilization, mathematical model of the bioreactor and on-line process monitoring.

Microbial Community Structure and Functions in Ethanol-Fed Sulfate Removal Bioreactors for Treatment of Mine Water

Directory of Open Access Journals (Sweden)

Malin Bomberg

2017-09-01

Full Text Available Sulfate-rich mine water must be treated before it is released into natural water bodies. We tested ethanol as substrate in bioreactors designed for biological sulfate removal from mine water containing up to 9 g L−1 sulfate, using granular sludge from an industrial waste water treatment plant as inoculum. The pH, redox potential, and sulfate and sulfide concentrations were measured twice a week over a maximum of 171 days. The microbial communities in the bioreactors were characterized by qPCR and high throughput amplicon sequencing. The pH in the bioreactors fluctuated between 5.0 and 7.7 with the highest amount of up to 50% sulfate removed measured around pH 6. Dissimilatory sulfate reducing bacteria (SRB constituted only between 1% and 15% of the bacterial communities. Predicted bacterial metagenomes indicated a high prevalence of assimilatory sulfate reduction proceeding to formation of l-cystein and acetate, assimilatory and dissimilatory nitrate reduction, denitrification, and oxidation of ethanol to acetaldehyde with further conversion to ethanolamine, but not to acetate. Despite efforts to maintain optimal conditions for biological sulfate reduction in the bioreactors, only a small part of the microorganisms were SRB. The microbial communities were highly diverse, containing bacteria, archaea, and fungi, all of which affected the overall microbial processes in the bioreactors. While it is important to monitor specific physicochemical parameters in bioreactors, molecular assessment of the microbial communities may serve as a tool to identify biological factors affecting bioreactor functions and to optimize physicochemical attributes for ideal bioreactor performance.

A comparative study of leachate quality and biogas generation in simulated anaerobic and hybrid bioreactors

Energy Technology Data Exchange (ETDEWEB)

Xu, Qiyong; Tian, Ying; Wang, Shen; Ko, Jae Hac, E-mail: jaehacko@pkusz.edu.cn

2015-07-15

Highlights: • Temporary aeration shortened the initial acid inhibition phase for methanogens. • COD decreased faster in the hybrid bioreactor than that in the anaerobic control. • Methane generations from hybrid bioreactors were 133.4 L/kg{sub vs} and 113.2 L/kg{sub vs}. • MSW settlement increased with increasing the frequency of intermittent aeration. - Abstract: Research has been conducted to compare leachate characterization and biogas generation in simulated anaerobic and hybrid bioreactor landfills with typical Chinese municipal solid waste (MSW). Three laboratory-scale reactors, an anaerobic (A1) and two hybrid bioreactors (C1 and C2), were constructed and operated for about 10 months. The hybrid bioreactors were operated in an aerobic–anaerobic mode with different aeration frequencies by providing air into the upper layer of waste. Results showed that the temporary aeration into the upper layer aided methane generation by shortening the initial acidogenic phase because of volatile fatty acids (VFAs) reduction and pH increase. Chemical oxygen demand (COD) decreased faster in the hybrid bioreactors, but the concentrations of ammonia–nitrogen in the hybrid bioreactors were greater than those in the anaerobic control. Methanogenic conditions were established within 75 d and 60 d in C1 and C2, respectively. However, high aeration frequency led to the consumption of organic matters by aerobic degradation and resulted in reducing accumulative methane volume. The temporary aeration enhanced waste settlement and the settlement increased with increasing the frequency of aeration. Methane production was inhibited in the anaerobic control; however, the total methane generations from hybrid bioreactors were 133.4 L/kg{sub vs} and 113.2 L/kg{sub vs}. As for MSW with high content of food waste, leachate recirculation right after aeration stopped was not recommended due to VFA inhibition for methanogens.

Characteristics of Brain Perfusion in Patients of Parkinson's Disease

International Nuclear Information System (INIS)

Jeong, Young Jin; Park, Min Jung; Kim, Jae Woo; Kang, Young Kang

2008-01-01

It was well known that cerebral blood perfusion is normal or diffusely decreased in the majority of patients with Parkinson's disease (PD). Actually we interpreted brain perfusion SPECT images of PD patients in the clinical situation, we observed various cerebral perfusion patterns in patients with PD. So we performed brain perfusion SPECT to know the brain perfusion patterns of PD patients and the difference of perfusion patterns according to the sex and the age. Also we classified PD patients into small groups based on the brain perfusion pattern. Two hundred nineteen patients (M: 70, F: 149, mean age: 62.9±6.9 y/o) who were diagnosed as PD without dementia clinically and 55 patients (M: 15, F: 40, mean age: 61.4±9.2 y/o) as normal controls who had no past illness history were performed 99m Tc-HMPAO brain perfusion SPECT and neuropsychological test. At first, we compared all patients with PD and normal controls. Brain perfusion in left inferior frontal gyrus, left insula, left transverse temporal gyrus, left inferior parietal lobule, left superior parietal lobule, right precuneus, right caudate tail were lower in patients with PD than normal controls. Secondly, we compared male and female patients with PD and normal controls, respectively. Brain perfusion SPECT showed more decreased cerebral perfusion in left hemisphere than right side in both male and female patients compared to normal controls. And there was larger hypoperfusion area in female patients compared with male. Thirdly, we classified patients with PD and normal controls into 4 groups according to the age and compared brain perfusion respectively. In patient below fifties, brain perfusion in both occipitoparietal and left temporal lobe were lower in PD group. As the patients with PD grew older, hypoperfusion area were shown in both frontal, temporal and limbic lobes. Fourthly, We were able to divide patients into small groups based on cerebral perfusion pattern. There was normal cerebral blood

Methyl Jasmonate and Salicylic Acid Induced Oxidative Stress and Accumulation of Phenolics in Panax ginseng Bioreactor Root Suspension Cultures

Directory of Open Access Journals (Sweden)

Kee-Yoeup Paek

2007-03-01

Full Text Available To investigate the enzyme variations responsible for the synthesis of phenolics, 40 day-old adventitious roots of Panax ginseng were treated with 200 μM methyl jasmonate (MJ or salicylic acid (SA in a 5 L bioreactor suspension culture (working volume 4 L. Both treatments caused an increase in the carbonyl and hydrogen peroxide (H2O2 contents, although the levels were lower in SA treated roots. Total phenolic, flavonoid, ascorbic acid, non-protein thiol (NPSH and cysteine contents and 1,1-diphenyl-2-picrylhydrazyl (DPPH radical reducing activity were increased by MJ and SA. Fresh weight (FW and dry weight (DW decreased significantly after 9 days of exposure to SA and MJ. The highest total phenolics (62%, DPPH activity (40%, flavonoids (88%, ascorbic acid (55%, NPSH (33%, and cysteine (62% contents compared to control were obtained after 9 days in SA treated roots. The activities of glucose 6-phosphate dehydrogenase, phenylalanine ammonia lyase, substrate specific peroxidases (caffeic acid peroxidase, quercetin peroxidase and ferulic acid peroxidase were higher in MJ treated roots than the SA treated ones. Increased shikimate dehydrogenase, chlorogenic acid peroxidase and β-glucosidase activities and proline content were observed in SA treated roots than in MJ ones. Cinnamyl alcohol dehydrogenase activity remained unaffected by both MJ and SA. These results strongly indicate that MJ and SA induce the accumulation of phenolic compounds in ginseng root by altering the phenolic synthesis enzymes.

Regional Cerebral Perfusion in Progressive Supranuclear Palsy

Energy Technology Data Exchange (ETDEWEB)

Lee, Won Yong; Lee, Ki Hyeong; Yoon, Byung Woo; Lee, Sang Bok; Jeon, Beom S. [Samsung Medical Center, Seoul (Korea, Republic of); Lee, Kyung Han; Lee, Myung Chul [Seoul National University College of Medicine, Seoul (Korea, Republic of)

1996-03-15

Progressive supranuclear palsy (PSP) is a Parkinson-plus syndrome characterized clinically by supranuclear ophthalmoplegia, pseudobulbar palsy, axial rigidity, bradykinesia, postural instability and dementia. Presence of dementia and lack of cortical histopathology suggest the derangement of cortical function by pathological changes in subcortical structures in PSP, which is supported by the pattern of behavioral changes and measurement of brain metabolism using positron emission tomography. This study was done to examine whether there are specific changes of regional cerebral perfusion in PSP and whether there is a correlation between severity of motor abnormaility and degree of changes in cerebral perfusion. We measured regional cerebral perfusion indices in 5 cortical and 2 subcortical areas in 6 patients with a clinical diagnosis of PSP and 6 healthy age and sex matched controls using Tc-99m-HMPAO SPECT. Compared with age and sex matched controls, only superior frontal regional perfusion index was significantly decreased in PSP (p<0.05). There was no correlation between the severity of the motor abnormality and any of the regional cerebral perfusion indices (p>0.05). We affirm the previous reports that perfusion in superior frontal cortex is decreased in PSP. Based on our results that there was no correlation between severity of motor abnormality and cerebral perfusion in the superior frontal cortex, nonmotoric symptoms including dementia needs to be looked at whether there is a correlation with the perfusion abnormality in superior frontal cortex

Dextran Utilization During Its Synthesis by Weissella cibaria RBA12 Can Be Overcome by Fed-Batch Fermentation in a Bioreactor.

Science.gov (United States)

Baruah, Rwivoo; Deka, Barsha; Kashyap, Niharika; Goyal, Arun

2018-01-01

Weissella cibaria RBA12 produced a maximum of 9 mg/ml dextran (with 90% efficiency) using shake flask culture under the optimized concentration of medium components viz. 2% (w/v) of each sucrose, yeast extract, and K 2 HPO 4 after incubation at optimized conditions of 20 °C and 180 rpm for 24 h. The optimized medium and conditions were used for scale-up of dextran production from Weissella cibaria RBA12 in 2.5-l working volume under batch fermentation in a bioreactor that yielded a maximum of 9.3 mg/ml dextran (with 93% efficiency) at 14 h. After 14 h, dextran produced was utilized by the bacterium till 18 h in its stationary phase under sucrose depleted conditions. Dextran utilization was further studied by fed-batch fermentation using sucrose feed. Dextran on production under fed-batch fermentation in bioreactor gave 35.8 mg/ml after 32 h. In fed-batch mode, there was no decrease in dextran concentration as observed in the batch mode. This showed that the utilization of dextran by Weissella cibaria RBA12 is initiated when there is sucrose depletion and therefore the presence of sucrose can possibly overcome the dextran hydrolysis. This is the first report of utilization of dextran, post-sucrose depletion by Weissella sp. studied in bioreactor.

A review of some parameters involved in fluidized bed bioreactors

Energy Technology Data Exchange (ETDEWEB)

Wright, P.C. [School of Chemical Engineering and Industrial Chemistry, The Univ. of New South Wales, Sydney (Australia); Raper, J.A. [School of Chemical Engineering and Industrial Chemistry, The Univ. of New South Wales, Sydney (Australia)

1996-02-01

Three-phase fluidized bed bioreactors have advantages over conventional chemical reaction systems. There is a lack of agreement over most major operational conditions, and a wide range of design variables are open to question. A large body of recent work in the field has been reviewed, with a degree of historical comparison and discussion. It has been found that aspects of fluidized bed biofilm reactors of vital importance include: choice of solid media, gas and liquid loadings, bacterial type and reactor mechanical design. A large proportion of the work in the field of three-phase fluidization is non-biologically specific, or not tested on a bacterially inoculated system. The majority of three-phase fluidized bed bioreactor work is in the field of water treatment. Although this work has highlighted the potential for use of bio-fluidized beds for this application, there are still specific problems hinderin the large scale industrial acceptance of three-phase fluidized bed bioreactors. (orig.)

Membrane bioreactors' potential for ethanol and biogas production: a review.

Science.gov (United States)

Ylitervo, Päivi; Akinbomia, Julius; Taherzadeha, Mohammad J

2013-01-01

Companies developing and producing membranes for different separation purposes, as well as the market for these, have markedly increased in numbers over the last decade. Membrane and separation technology might well contribute to making fuel ethanol and biogas production from lignocellulosic materials more economically viable and productive. Combining biological processes with membrane separation techniques in a membrane bioreactor (MBR) increases cell concentrations extensively in the bioreactor. Such a combination furthermore reduces product inhibition during the biological process, increases product concentration and productivity, and simplifies the separation of product and/or cells. Various MBRs have been studied over the years, where the membrane is either submerged inside the liquid to be filtered, or placed in an external loop outside the bioreactor. All configurations have advantages and drawbacks, as reviewed in this paper. The current review presents an account of the membrane separation technologies, and the research performed on MBRs, focusing on ethanol and biogas production. The advantages and potentials of the technology are elucidated.

Regional Cerebral Perfusion in Progressive Supranuclear Palsy

International Nuclear Information System (INIS)

Lee, Won Yong; Lee, Ki Hyeong; Yoon, Byung Woo; Lee, Sang Bok; Jeon, Beom S.; Lee, Kyung Han; Lee, Myung Chul

1996-01-01

Progressive supranuclear palsy (PSP) is a Parkinson-plus syndrome characterized clinically by supranuclear ophthalmoplegia, pseudobulbar palsy, axial rigidity, bradykinesia, postural instability and dementia. Presence of dementia and lack of cortical histopathology suggest the derangement of cortical function by pathological changes in subcortical structures in PSP, which is supported by the pattern of behavioral changes and measurement of brain metabolism using positron emission tomography. This study was done to examine whether there are specific changes of regional cerebral perfusion in PSP and whether there is a correlation between severity of motor abnormaility and degree of changes in cerebral perfusion. We measured regional cerebral perfusion indices in 5 cortical and 2 subcortical areas in 6 patients with a clinical diagnosis of PSP and 6 healthy age and sex matched controls using Tc-99m-HMPAO SPECT. Compared with age and sex matched controls, only superior frontal regional perfusion index was significantly decreased in PSP (p 0.05). We affirm the previous reports that perfusion in superior frontal cortex is decreased in PSP. Based on our results that there was no correlation between severity of motor abnormality and cerebral perfusion in the superior frontal cortex, nonmotoric symptoms including dementia needs to be looked at whether there is a correlation with the perfusion abnormality in superior frontal cortex

Computerized analysis of brain perfusion parameter images

International Nuclear Information System (INIS)

Turowski, B.; Haenggi, D.; Wittsack, H.J.; Beck, A.; Aurich, V.

2007-01-01

Purpose: The development of a computerized method which allows a direct quantitative comparison of perfusion parameters. The display should allow a clear direct comparison of brain perfusion parameters in different vascular territories and over the course of time. The analysis is intended to be the basis for further evaluation of cerebral vasospasm after subarachnoid hemorrhage (SAH). The method should permit early diagnosis of cerebral vasospasm. Materials and Methods: The Angiotux 2D-ECCET software was developed with a close cooperation between computer scientists and clinicians. Starting from parameter images of brain perfusion, the cortex was marked, segmented and assigned to definite vascular territories. The underlying values were averages for each segment and were displayed in a graph. If a follow-up was available, the mean values of the perfusion parameters were displayed in relation to time. The method was developed under consideration of CT perfusion values but is applicable for other methods of perfusion imaging. Results: Computerized analysis of brain perfusion parameter images allows an immediate comparison of these parameters and follow-up of mean values in a clear and concise manner. Values are related to definite vascular territories. The tabular output facilitates further statistic evaluations. The computerized analysis is precisely reproducible, i. e., repetitions result in exactly the same output. (orig.)

Aortic valve biomechanics during LVAD support: Implementation in a bioreactor design and preliminary testing

Science.gov (United States)

Jamal, Madiha

Although Left Ventricle Assist Device (LVAD) support has enhanced the quality of life for many heart failure patients, its prolonged in-vivo implantation causes change in hemodynamics and biomechanics of the aortic heart valve eventually leading to development of aortic insufficiency (AI). The LVAD decreases pressure in the left ventricle, leading to high transvalvular pressure and reduced aortic valve opening. Our hypothesis is that the increased pressure leads to increased mechanical stretch in the aortic valve leaflets, inducing a cascade of responses that ultimately result in local tissue fibrosis and AI. The goal of this study is to investigate the mechanism behind this hypothesis using the methods of tissue engineering. A bioreactor has been built that imparts cyclic stretch and flow to small 3-D constructs of living cells cultured in a silicone membrane. The approach is to use this device for in vitro tissue culture of vascular interstitial cells (VICs) embedded in a collagen gel, which will be subjected to normal and altered stretch and shear representative of the in vivo valve biomechanics. The bioreactor was validated to measure the amount of stretch and shear it can impart to closely replicate in-vivo conditions using PIV technique and ImageJ software. Mean longitudinal strain of 0.037cm (SD= +/-0.013cm) was recorded with mean perpendicular strain being 0.0046cm (SD= +/-0.0169cm). Measured average shear stress imparted at 100ml/min was 2.735 dynes/cm2 (SD= +/-2.25 dynes/cm2) with 6.21 dynes/cm2 (SD= +/-3.35 dynes/cm2) at 200ml/min. The cells that underwent cycles of stretch and shear in the bioreactor were screened for formation of myofibroblast using techniques of immunohistochemistry. The marker used was ? smooth muscle actin (SMA) which identifies pathological differentiation of the CPCs to myofibroblast. Yhe expression of the myofibroblast phenotype is a feature of valvupathy. In case of Shear Vs Static control, the mean value for SMA expression for

Novel bacterial sulfur oxygenase reductases from bioreactors treating gold-bearing concentrates

DEFF Research Database (Denmark)

Chen, Z-W; Liu, Y-Y; Wu, J-F

2007-01-01

The microbial community and sulfur oxygenase reductases of metagenomic DNA from bioreactors treating gold-bearing concentrates were studied by 16S rRNA library, real-time polymerase chain reaction (RT-PCR), conventional cultivation, and molecular cloning. Results indicated that major bacterial......) of bacteria and archaea were 4.59 x 10(9) and 6.68 x 10(5), respectively. Bacterial strains representing Acidithiobacillus, Leptospirillum, and Sulfobacillus were isolated from the bioreactors. To study sulfur oxidation in the reactors, pairs of new PCR primers were designed for the detection of sulfur...... oxygenase reductase (SOR) genes. Three sor-like genes, namely, sor (Fx), sor (SA), and sor (SB) were identified from metagenomic DNAs of the bioreactors. The sor (Fx) is an inactivated SOR gene and is identical to the pseudo-SOR gene of Ferroplasma acidarmanus. The sor (SA) and sor (SB) showed...

Comparison with myocardial perfusion MRI and myocardial perfusion SPECT in the diagnostic performance of coronary artery disease. A meta-analysis

International Nuclear Information System (INIS)

Iwata, Kunihiro; Kubota, Makoto; Ogasawara, Katsuhiko

2008-01-01

We compared the diagnostic abilities of stress myocardial perfusion MRI (myocardial perfusion MRI) and myocardial perfusion single photon emission computed tomography (SPECT), using a meta-analysis method. We investigated the diagnostic abilities of MRI and SPECT in similar subject groups in reports written in English or Japanese. The reports to be used for analysis were selected according to a ''screening standard,'' which was established in advance. After consolidating the data from the selected reports, we compared the integrated odds ratio, the point estimation values of sensibility/specificity, and the summary receiver operating characteristic (ROC) curve. For the analysis, six reports were selected (subjects: 153, coronary-artery target sites: 447). Meta-analysis revealed that the diagnostic ability of myocardial perfusion MRI was superior to that of myocardial perfusion SPECT regarding each of the parameters. This is considered to be supportive evidence of the usefulness of myocardial perfusion MRI. (author)

Tc-99m DTPA perfusion scintigraphy and color coded duplex sonography in the evaluation of minimal renal allograft perfusion

International Nuclear Information System (INIS)

Bair, H.J.; Platsch, G.; Wolf, F.; Guenter, E.; Becker, D.; Rupprecht, H.; Neumayer, H.H.

1997-01-01

Aim: The clinical impact of perfusion scintigraphy versus color coded Duplex sonography was evaluated, with respect to their potential in assessing minimal allograft perfusion in vitally threatened kidney transplants, i.e. oligoanuric allografts suspected to have either severe rejection or thrombosis of the renal vein or artery. Methods: From July 1990 to August 1994 the grafts of 15 out of a total of 315 patients were vitally threatened. Technetium-99m DTPA scintigraphy and color coded Duplex sonography were performed in all patients. For scintigraphic evaluation of transplant perfusion analog scans up to 60 min postinjection, and time-activity curves over the first 60 sec after injection of 370-440 MBq Tc-99m diethylenetriaminepentaacetate acid (DTPA) were used and classified by a perfusion score, the time between renal and iliac artery peaks (TDiff) and the washout of the renogram curve. Additionally, evaluation of excretion function and assessment of vascular or urinary leaks were performed. By color coded Duplex sonography the perfusion in all sections of the graft as well as the vascular anastomoses were examined and the maximal blood flow velocity (Vmax) and the resistive index (RI) in the renal artery were determined by means of the pulsed Doppler device. Pathologic-anatomical diagnosis was achieved by either biopsy or post-explant histology in all grafts. Results: Scintigraphy and color coded Duplex sonography could reliably differentiate minimal (8/15) and not perfused (7/15) renal allografts. The results were confirmed either by angiography in digital subtraction technique (DSA) or the clinical follow up. Conclusion: In summary, perfusion scintigraphy and color coded Duplex sonography are comparable modalities to assess kidney graft perfusion. In clinical practice scintigraphy and colorcoded Doppler sonography can replace digital subtraction angiography in the evaluation of minimal allograft perfusion. (orig.) [de

Normal anatomy of lung perfusion SPECT scintigraphy

International Nuclear Information System (INIS)

Moskowitz, G.W.; Levy, L.M.

1987-01-01

Ten patients studies for possible pulmonary embolic disease had normal lung perfusion planar and SPECT scintigraphy. A computer program was developed to superimpose the CT scans on corresponding SPECT images. Superimposition of CT scans on corresponding SPECT transaxial cross-sectional images, when available, provides the needed definition and relationships of adjacent organs. SPECT transaxial sections provide clear anatomic definition of perfusion defects without foreground and background lung tissue superimposed. The location, shape, and size of the perfusion defects can be readily assessed by SPECT. An algorithm was developed for the differentiation of abnormal pulmonary perfusion patterns from normal structures on variation

Air purification from TCE and PCE contamination in a hybrid bioreactors and biofilter integrated system.

Science.gov (United States)

Tabernacka, Agnieszka; Zborowska, Ewa; Lebkowska, Maria; Borawski, Maciej

2014-01-15

A two-stage waste air treatment system, consisting of hybrid bioreactors (modified bioscrubbers) and a biofilter, was used to treat waste air containing chlorinated ethenes - trichloroethylene (TCE) and tetrachloroethylene (PCE). The bioreactor was operated with loadings in the range 0.46-5.50gm(-3)h(-1) for TCE and 2.16-9.02gm(-3)h(-1) for PCE. The biofilter loadings were in the range 0.1-0.97gm(-3)h(-1) for TCE and 0.2-2.12gm(-3)h(-1) for PCE. Under low pollutant loadings, the efficiency of TCE elimination was 23-25% in the bioreactor and 54-70% in the biofilter. The efficiency of PCE elimination was 44-60% in the bioreactor and 50-75% in the biofilter. The best results for the bioreactor were observed one week after the pollutant loading was increased. However, the process did not stabilize. In the next seven days contaminant removal efficiency, enzymatic activity and biomass content were all diminished. Copyright © 2013 Elsevier B.V. All rights reserved.

Effects of granular activated carbon on methane removal performance and methanotrophic community of a lab-scale bioreactor.

Science.gov (United States)

Lee, Eun-Hee; Choi, Sun-Ah; Yi, Taewoo; Kim, Tae Gwan; Lee, Sang-Don; Cho, Kyung-Suk

2015-01-01

Two identical lab-scale bioreactor systems were operated to examine the effects of granular activated carbon (GAC) on methane removal performance and methanotrophic community. Both bioreactor systems removed methane completely at a CH4 loading rate of 71.2 g-CH4·d(-1) for 17 days. However, the methane removal efficiency declined to 88% in the bioreactor without GAC, while the bioreactor amended with GAC showed greater methane removal efficiency of 97% at a CH4 loading rate of 107.5 g-CH4·d(-1). Although quantitative real-time PCR showed that methanotrophic populations were similar levels of 5-10 × 10(8) pmoA gene copy number·VSS(-1) in both systems, GAC addition changed the methanotrophic community composition of the bioreactor systems. Microarray assay revealed that GAC enhanced the type I methanotrophic genera including Methylobacter, Methylomicrobium, and Methylomonas of the system, which suggests that GAC probably provided a favorable environment for type I methanotrophs. These results indicated that GAC is a promising support material in bioreactor systems for CH4 mitigation.

Regional cortical hyper perfusion on perfusion CT during postical motor deficit: A case report

Energy Technology Data Exchange (ETDEWEB)

Baik, Hye Jin [Dept. of Radiology, Haeundae Paik Hospital, Inje University College of Medicine, Busan (Korea, Republic of)

2013-08-15

Postictal neurologic deficit is a well-known complication mimicking the manifestation of a stroke. We present a case of a patient with clinical evidence of Todd's paralysis correlating with reversible postictal parenchymal changes on perfusion CT and magnetic resonance (MR) imaging. In this case, perfusion CT and MR imaging were helpful in the differential diagnosis of stroke-mimicking conditions.

Wastewater treatments by membrane bioreactors (MBR); Bioreactores de membrana (MBR) para la depuracion de aguas residuales

Energy Technology Data Exchange (ETDEWEB)

Guardino Ferre, R.

2001-07-01

Wastewater treatments by membrane bioreactors (MBR), are a good alternative of treatment to the conventional processes when wish to obtain very high quality of the treated water or to try high load contaminants in low flow. Simultaneously, the article explains the significant reduction of the wastewater treatment plant space, eliminating the secondary septic tank. (Author) 7 refs.

Ventilation and perfusion display in a single image

International Nuclear Information System (INIS)

Lima, J.J.P. de; Botelho, M.F.R.; Pereira, A.M.S.; Rafael, J.A.S.; Pinto, A.J.; Marques, M.A.T.; Pereira, M.C.; Baganha, M.F.; Godinho, F.

1991-01-01

A new method of ventilation and perfusion display onto a single image is presented. From the data on regions of interest of the lungs, three-dimensional histograms are created, containing as parameters X and Y for the position of the pixels, Z for the perfusion and colour for local ventilation. The perfusion value is supplied by sets of curves having Z proportional to the local perfusion count rate. Ventilation modulates colour. Four perspective views of the histogram are simultaneously displayed to allow visualization of the entire organ. Information about the normal ranges for both ventilation and perfusion is also provided in the histograms. (orig.)

Manufacturing recombinant proteins in kg-ton quantities using animal cells in bioreactors.

Science.gov (United States)

De Jesus, Maria; Wurm, Florian M

2011-06-01

Mammalian cells in bioreactors as production host are the focus of this review. We wish to briefly describe today's technical status and to highlight emerging trends in the manufacture of recombinant therapeutic proteins, focusing on Chinese hamster ovary (CHO) cells. CHO cells are the manufacturing host system of choice for more than 70% of protein pharmaceuticals on the market [21]. The current global capacity to grow mammalian cells in bioreactors stands at about 0.5 million liters, whereby the largest vessels can have a working volume of about 20,000l. We are focusing in this article on the upstream part of protein manufacturing. Over the past 25 years, volumetric yields for recombinant cell lines have increased about 20-fold mainly as the result of improvements in media and bioprocess design. Future yield increases are expected to come from improved gene delivery methods, from improved, possibly genetically modified host systems, and from further improved bioprocesses in bioreactors. Other emerging trends in protein manufacturing that are discussed include the use of disposal bioreactors and transient gene expression. We specifically highlight here current research in our own laboratories. Copyright © 2011 Elsevier B.V. All rights reserved.

Membrane filtration device for studying compression of fouling layers in membrane bioreactors.

Directory of Open Access Journals (Sweden)

Mads Koustrup Jørgensen

Full Text Available A filtration devise was developed to assess compressibility of fouling layers in membrane bioreactors. The system consists of a flat sheet membrane with air scouring operated at constant transmembrane pressure to assess the influence of pressure on resistance of fouling layers. By fitting a mathematical model, three model parameters were obtained; a back transport parameter describing the kinetics of fouling layer formation, a specific fouling layer resistance, and a compressibility parameter. This stands out from other on-site filterability tests as model parameters to simulate filtration performance are obtained together with a characterization of compressibility. Tests on membrane bioreactor sludge showed high reproducibility. The methodology's ability to assess compressibility was tested by filtrations of sludges from membrane bioreactors and conventional activated sludge wastewater treatment plants from three different sites. These proved that membrane bioreactor sludge showed higher compressibility than conventional activated sludge. In addition, detailed information on the underlying mechanisms of the difference in fouling propensity were obtained, as conventional activated sludge showed slower fouling formation, lower specific resistance and lower compressibility of fouling layers, which is explained by a higher degree of flocculation.

Bioreactors

Energy Technology Data Exchange (ETDEWEB)

Jamaleddine, E. [McGill Univ., Montreal, PQ (Canada). Dept. of Bioresource Engineering

2010-07-01

Composting is once again gaining interest among ecological engineers in view of greener industrial and residential activities. Uniform composting is needed to ensure decomposition and to keep the whole system at the same composting stage. A homogeneous temperature must be maintained throughout the media. A bioreactor design consisting of a heater core made of copper tubing was designed and tested. Two four-inch holes were made at the top and bottom of the barrel to allow air to flow through the system and promote aerobic composting. Once composting began and temperature increased, the water began to flow through the copper piping and the core heat was distributed throughout the medium. Three thermocouples were inserted at different heights on a 200 litre plastic barrel fitted with the aforementioned apparatus. Temperature variations were found to be considerably lower when the apparatus was operated with the heat redistribution system, enabling uniform composting, accelerating the process and reducing the risks of pathogenic or other contaminants remaining active in the barrels.

Mycelium differentiation and development of Streptomyces coelicolor in lab-scale bioreactors: Programmed cell death, differentiation, and lysis are closely linked to undecylprodigiosin and actinorhodin production

OpenAIRE

Rioseras de Bustos, Beatriz; López García, María Teresa (Bio); Yagüe Menéndez, Paula; Sánchez Martín, Jesús; Manteca Fernández, Ángel

2014-01-01

Streptomycetes are mycelium-forming bacteria that produce two thirds of clinically relevant secondary metabolites. Secondary metabolite production is activated at specific developmental stages of Streptomyces life cycle. Despite this, Streptomyces differentiation in industrial bioreactors tends to be underestimated and the most important parameters managed are only indirectly related to differentiation: modifications to the culture media, optimization of productive strains by random or direct...

ANAEROBIC MEMBRANE BIOREACTORS FOR DOMESTIC WASTEWATER TREATMENT. PRELIMINARY STUDY

Directory of Open Access Journals (Sweden)

Luisa Vera

2014-12-01

Full Text Available The operation of submerged anaerobic membrane bioreactors (SAnMBRs for domestic wastewaters treatment was studied in laboratory scale, with the objective to define sustainable filtration conditions of the suspensions along the process. During continuous experiments, the organic matter degradation by anaerobic way showed an average DQOT removal of 85% and 93%. Indeed, the degradation generated biogas after 12 days of operation and its relative methane composition was of 60% after 25 days of operation. Additionally, the comparison between membrane bioreactors (MBRs performance in aerobic and anaerobic conditions in filterability terms, reported that both systems behave similarly once reached the stationary state.

Effects of Steroid Hormones on Sex Differences in Cerebral Perfusion.

Directory of Open Access Journals (Sweden)

Carmen Ghisleni

Full Text Available Sex differences in the brain appear to play an important role in the prevalence and progression of various neuropsychiatric disorders, but to date little is known about the cerebral mechanisms underlying these differences. One widely reported finding is that women demonstrate higher cerebral perfusion than men, but the underlying cause of this difference in perfusion is not known. This study investigated the putative role of steroid hormones such as oestradiol, testosterone, and dehydroepiandrosterone sulphate (DHEAS as underlying factors influencing cerebral perfusion. We acquired arterial spin labelling perfusion images of 36 healthy adult subjects (16 men, 20 women. Analyses on average whole brain perfusion levels included a multiple regression analysis to test for the relative impact of each hormone on the global perfusion. Additionally, voxel-based analyses were performed to investigate the sex difference in regional perfusion as well as the correlations between local perfusion and serum oestradiol, testosterone, and DHEAS concentrations. Our results replicated the known sex difference in perfusion, with women showing significantly higher global and regional perfusion. For the global perfusion, DHEAS was the only significant predictor amongst the steroid hormones, showing a strong negative correlation with cerebral perfusion. The voxel-based analyses revealed modest sex-dependent correlations between local perfusion and testosterone, in addition to a strong modulatory effect of DHEAS in cortical, subcortical, and cerebellar regions. We conclude that DHEAS in particular may play an important role as an underlying factor driving the difference in cerebral perfusion between men and women.

CT perfusion of the liver during selective hepatic arteriography. Pure arterial blood perfusion of liver tumor and parenchyma

International Nuclear Information System (INIS)

Komemushi, Atsushi; Tanigawa, Noboru; Kojima, Hiroyuki; Kariya, Shuji; Sawada, Satoshi

2003-01-01

The purpose of this study was to quantify pure arterial blood perfusion of liver tumor and parenchyma by using CT perfusion during selective hepatic arteriography. A total of 44 patients underwent liver CT perfusion study by injection of contrast medium via the hepatic artery. CT-perfusion parameters including arterial blood flow, arterial blood volume, and arterial mean transit time in the liver parenchyma and liver tumor were calculated using the deconvolution method. The CT-perfusion parameters and vascularity of the tumor were compared. A complete analysis could be performed in 36 of the 44 patients. For liver tumor and liver parenchyma, respectively, arterial blood flow was 184.6±132.7 and 41.0±27.0 ml/min/100 g, arterial blood volume was 19.4±14.6 and 4.8±4.2 ml/100 g, and arterial mean transit time was 8.9±4.2 and 10.2±5.3 sec. Arterial blood flow and arterial blood volume correlated significantly with the vascularity of the tumor; however no correlation was detected between arterial mean transit time and the vascularity of the tumor. This technique could be used to quantify pure hepatic arterial blood perfusion. (author)

Quantitative perfusion imaging in magnetic resonance imaging

International Nuclear Information System (INIS)

Zoellner, F.G.; Gaa, T.; Zimmer, F.; Ong, M.M.; Riffel, P.; Hausmann, D.; Schoenberg, S.O.; Weis, M.

2016-01-01

Magnetic resonance imaging (MRI) is recognized for its superior tissue contrast while being non-invasive and free of ionizing radiation. Due to the development of new scanner hardware and fast imaging techniques during the last decades, access to tissue and organ functions became possible. One of these functional imaging techniques is perfusion imaging with which tissue perfusion and capillary permeability can be determined from dynamic imaging data. Perfusion imaging by MRI can be performed by two approaches, arterial spin labeling (ASL) and dynamic contrast-enhanced (DCE) MRI. While the first method uses magnetically labelled water protons in arterial blood as an endogenous tracer, the latter involves the injection of a contrast agent, usually gadolinium (Gd), as a tracer for calculating hemodynamic parameters. Studies have demonstrated the potential of perfusion MRI for diagnostics and also for therapy monitoring. The utilization and application of perfusion MRI are still restricted to specialized centers, such as university hospitals. A broad application of the technique has not yet been implemented. The MRI perfusion technique is a valuable tool that might come broadly available after implementation of standards on European and international levels. Such efforts are being promoted by the respective professional bodies. (orig.) [de

Perfusion-weighted MR imaging of uterine leiomyoma

Energy Technology Data Exchange (ETDEWEB)

Takase, Hiroyasu; Munechika, Hirotsugu [Showa Univ., Tokyo (Japan). School of Medicine

2001-06-01

Serial images of uterine leiomyoma in gradient-echo, echo-planar, magnetic resonance imaging were taken to draw a {delta}R2{sup *} curve after intravenous bolus injection of Gd-DTPA. The {delta}R2{sup *} integral was calculated from a {delta}R2{sup *} curve to have relative perfusion of uterine leiomyoma. We then, evaluated the amount of perfusion correlated with MR findings, size and number of leiomyoma or the clinical symptoms and established that perfusion was correlated positively with the findings of T2 weighted images and clinical symptoms but not with other MR findings or size and number of leiomyoma. In conclusion, we presumed that the clinical symptoms could be reduced by decreasing of an amount of perfusion of uterine leiomyoma in some means. However, it remained uncertain why severe clinical symptoms were associated with a high amount of perfusion in uterine leiomyomas. (author)

Biological manganese removal from acid mine drainage in constructed wetlands and prototype bioreactors.

Science.gov (United States)

Hallberg, Kevin B; Johnson, D Barrie

2005-02-01

Mine drainage waters vary considerably in the range and concentration of heavy metals they contain. Besides iron, manganese is frequently present at elevated concentrations in waters draining both coal and metal mines. Passive treatment systems (aerobic wetlands and compost bioreactors) are designed to remove iron by biologically induced oxidation/precipitation. Manganese, however, is problematic as it does not readily form sulfidic minerals and requires elevated pH (>8) for abiotic oxidation of Mn (II) to insoluble Mn (IV). As a result, manganese removal in passive remediation systems is often less effective than removal of iron. This was found to be the case at the pilot passive treatment plant (PPTP) constructed to treat water draining the former Wheal Jane tin mine in Cornwall, UK, where effective removal of manganese occurred only in one of the three rock filter components of the composite systems over a 1-year period of monitoring. Water in the two rock filter systems where manganese removal was relatively poor was generally system. These differences in water chemistry and manganese removal were due to variable performances in the compost bioreactors that feed the rock filter units in the composite passive systems at Wheal Jane. An alternative approach for removing soluble manganese from mine waters, using fixed bed bioreactors, was developed. Ferromanganese nodules (about 2 cm diameter), collected from an abandoned mine adit in north Wales, were used to inoculate the bioreactors (working volume ca. 700 ml). Following colonization by manganese-oxidizing microbes, the aerated bioreactor catalysed the removal of soluble manganese, via oxidation of Mn (II) and precipitation of the resultant Mn (IV) in the bioreactor, in synthetic media and mine water from the Wheal Jane PPTP. Such an approach has potential application for removing soluble Mn from mine streams and other Mn-contaminated water courses.