WorldWideScience

Sample records for bioreactor culture model

  1. NASA Bioreactor tissue culture

    Science.gov (United States)

    1998-01-01

    Dr. Lisa E. Freed of the Massachusetts Institute of Technology and her colleagues have reported that initially disc-like specimens tend to become spherical in space, demonstrating that tissues can grow and differentiate into distinct structures in microgravity. The Mir Increment 3 (Sept. 16, 1996 - Jan. 22, 1997) samples were smaller, more spherical, and mechanically weaker than Earth-grown control samples. These results demonstrate the feasibility of microgravity tissue engineering and may have implications for long human space voyages and for treating musculoskeletal disorders on earth. 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.

  2. NASA Bioreactor tissue culture

    Science.gov (United States)

    1998-01-01

    Dr. Lisa E. Freed of the Massachusetts Institute of Technology and her colleagues have reported that initially disc-like specimens tend to become spherical in space, demonstrating that tissues can grow and differentiate into distinct structures in microgravity. The Mir Increment 3 (Sept. 16, 1996 - Jan. 22, 1997) samples were smaller, more spherical, and mechanically weaker than Earth-grown control samples. These results demonstrate the feasibility of microgravity tissue engineering and may have implications for long human space voyages and for treating musculoskeletal disorders on earth. 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.

  3. Oscillating Cell Culture Bioreactor

    Science.gov (United States)

    Freed, Lisa E.; Cheng, Mingyu; Moretti, Matteo G.

    2010-01-01

    To better exploit the principles of gas transport and mass transport during the processes of cell seeding of 3D scaffolds and in vitro culture of 3D tissue engineered constructs, the oscillatory cell culture bioreactor provides a flow of cell suspensions and culture media directly through a porous 3D scaffold (during cell seeding) and a 3D construct (during subsequent cultivation) within a highly gas-permeable closed-loop tube. This design is simple, modular, and flexible, and its component parts are easy to assemble and operate, and are inexpensive. Chamber volume can be very low, but can be easily scaled up. This innovation is well suited to work with different biological specimens, particularly with cells having high oxygen requirements and/or shear sensitivity, and different scaffold structures and dimensions. The closed-loop changer is highly gas permeable to allow efficient gas exchange during the cell seeding/culturing process. A porous scaffold, which may be seeded with cells, is fixed by means of a scaffold holder to the chamber wall with scaffold/construct orientation with respect to the chamber determined by the geometry of the scaffold holder. A fluid, with/without biological specimens, is added to the chamber such that all, or most, of the air is displaced (i.e., with or without an enclosed air bubble). Motion is applied to the chamber within a controlled environment (e.g., oscillatory motion within a humidified 37 C incubator). Movement of the chamber induces relative motion of the scaffold/construct with respect to the fluid. In case the fluid is a cell suspension, cells will come into contact with the scaffold and eventually adhere to it. Alternatively, cells can be seeded on scaffolds by gel entrapment prior to bioreactor cultivation. Subsequently, the oscillatory cell culture bioreactor will provide efficient gas exchange (i.e., of oxygen and carbon dioxide, as required for viability of metabolically active cells) and controlled levels of fluid

  4. Use of microgravity bioreactors for development of an in vitro rat salivary gland cell culture model

    Science.gov (United States)

    Lewis, M. L.; Moriarity, D. M.; Campbell, P. S.

    1993-01-01

    During development, salivary gland (SG) cells both secrete factors which modulate cellular behavior and express specific hormone receptors. Whether SG cell growth is modulated by an autocrine epidermal growth factor (EGF) receptor-mediated signal transduction pathway is not clearly understood. SG tissue is the synthesis site for functionally distinct products including growth factors, digestive enzymes, and homeostasis maintaining factors. Historically, SG cells have proven difficult to grow and may be only maintained as limited three-dimensional ductal-type structures in collagen gels or on reconstituted basement membrane gels. A novel approach to establishing primary rat SG cultures is use of microgravity bioreactors originally designed by NASA as low-shear culture systems for predicting cell growth and differentiation in the microgravity environment of space. These completely fluid-filled bioreactors, which are oriented horizontally and rotate, have proven advantageous for Earth-based culture of three-dimensional cell assemblies, tissue-like aggregates, and glandular structures. Use of microgravity bioreactors for establishing in vitro models to investigate steroid-mediated secretion of EGF by normal SG cells may also prove useful for the investigation of cancer and other salivary gland disorders. These microgravity bioreactors promise challenging opportunities for future applications in basic and applied cell research.

  5. Culturing and applications of rotating wall vessel bioreactor derived 3D epithelial cell models.

    Science.gov (United States)

    Radtke, Andrea L; Herbst-Kralovetz, Melissa M

    2012-04-03

    Cells and tissues in the body experience environmental conditions that influence their architecture, intercellular communications, and overall functions. For in vitro cell culture models to accurately mimic the tissue of interest, the growth environment of the culture is a critical aspect to consider. Commonly used conventional cell culture systems propagate epithelial cells on flat two-dimensional (2-D) impermeable surfaces. Although much has been learned from conventional cell culture systems, many findings are not reproducible in human clinical trials or tissue explants, potentially as a result of the lack of a physiologically relevant microenvironment. Here, we describe a culture system that overcomes many of the culture condition boundaries of 2-D cell cultures, by using the innovative rotating wall vessel (RWV) bioreactor technology. We and others have shown that organotypic RWV-derived models can recapitulate structure, function, and authentic human responses to external stimuli similarly to human explant tissues (1-6). The RWV bioreactor is a suspension culture system that allows for the growth of epithelial cells under low physiological fluid shear conditions. The bioreactors come in two different formats, a high-aspect rotating vessel (HARV) or a slow-turning lateral vessel (STLV), in which they differ by their aeration source. Epithelial cells are added to the bioreactor of choice in combination with porous, collagen-coated microcarrier beads (Figure 1A). The cells utilize the beads as a growth scaffold during the constant free fall in the bioreactor (Figure 1B). The microenvironment provided by the bioreactor allows the cells to form three-dimensional (3-D) aggregates displaying in vivo-like characteristics often not observed under standard 2-D culture conditions (Figure 1D). These characteristics include tight junctions, mucus production, apical/basal orientation, in vivo protein localization, and additional epithelial cell-type specific properties

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

    Science.gov (United States)

    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 CO2 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

  7. Generic Raman-based calibration models enabling real-time monitoring of cell culture bioreactors.

    Science.gov (United States)

    Mehdizadeh, Hamidreza; Lauri, David; Karry, Krizia M; Moshgbar, Mojgan; Procopio-Melino, Renee; Drapeau, Denis

    2015-01-01

    Raman-based multivariate calibration models have been developed for real-time in situ monitoring of multiple process parameters within cell culture bioreactors. Developed models are generic, in the sense that they are applicable to various products, media, and cell lines based on Chinese Hamster Ovarian (CHO) host cells, and are scalable to large pilot and manufacturing scales. Several batches using different CHO-based cell lines and corresponding proprietary media and process conditions have been used to generate calibration datasets, and models have been validated using independent datasets from separate batch runs. All models have been validated to be generic and capable of predicting process parameters with acceptable accuracy. The developed models allow monitoring multiple key bioprocess metabolic variables, and hence can be utilized as an important enabling tool for Quality by Design approaches which are strongly supported by the U.S. Food and Drug Administration.

  8. Application of bioreactor design principles and multivariate analysis for development of cell culture scale down models.

    Science.gov (United States)

    Tescione, Lia; Lambropoulos, James; Paranandi, Madhava Ram; Makagiansar, Helena; Ryll, Thomas

    2015-01-01

    A bench scale cell culture model representative of manufacturing scale (2,000 L) was developed based on oxygen mass transfer principles, for a CHO-based process producing a recombinant human protein. Cell culture performance differences across scales are characterized most often by sub-optimal performance in manufacturing scale bioreactors. By contrast in this study, reduced growth rates were observed at bench scale during the initial model development. Bioreactor models based on power per unit volume (P/V), volumetric mass transfer coefficient (kL a), and oxygen transfer rate (OTR) were evaluated to address this scale performance difference. Lower viable cell densities observed for the P/V model were attributed to higher sparge rates and reduced oxygen mass transfer efficiency (kL a) of the small scale hole spargers. Increasing the sparger kL a by decreasing the pore size resulted in a further decrease in growth at bench scale. Due to sensitivity of the cell line to gas sparge rate and bubble size that was revealed by the P/V and kL a models, an OTR model based on oxygen enrichment and increased P/V was selected that generated endpoint sparge rates representative of 2,000 L scale. This final bench scale model generated similar growth rates as manufacturing. In order to take into account other routinely monitored process parameters besides growth, a multivariate statistical approach was applied to demonstrate validity of the small scale model. After the model was selected based on univariate and multivariate analysis, product quality was generated and verified to fall within the 95% confidence limit of the multivariate model.

  9. Bioreactor

    Science.gov (United States)

    1996-01-01

    The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. 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. 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 currently being cultured in rotating bioreactors by investigators

  10. Optimizing of Culture Conditionin Horizontal Rotating Bioreactor

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    1 IntroductionBioreactor is the most important equipment in tissue engineering. It can mimic the micro-environment of cell growth in vitro. At present, horizontal rotating bioreactor is the most advanced equipment for cell culture in the world. 2 Rotating bioreactors2.1 Working principleThere are two kinds of horizontal rotating bioreactor: HARV(high aspect ratio vessel) and RCCS (rotary cell culture system). It is drived by step motor with horizontal rotation, the culture medium and cell is filled between ...

  11. Hairy root culture: bioreactor design and process intensification.

    Science.gov (United States)

    Stiles, Amanda R; Liu, Chun-Zhao

    2013-01-01

    The cultivation of hairy roots for the production of secondary metabolites offers numerous advantages; hairy roots have a fast growth rate, are genetically stable, and are relatively simple to maintain in phytohormone free media. Hairy roots provide a continuous source of secondary metabolites, and are useful for the production of chemicals for pharmaceuticals, cosmetics, and food additives. In order for hairy roots to be utilized on a commercial scale, it is necessary to scale-up their production. Over the last several decades, significant research has been conducted on the cultivation of hairy roots in various types of bioreactor systems. In this review, we discuss the advantages and disadvantages of various bioreactor systems, the major factors related to large-scale bioreactor cultures, process intensification technologies and overview the mathematical models and computer-aided methods that have been utilized for bioreactor design and development.

  12. Human cell culture in a space bioreactor

    Science.gov (United States)

    Morrison, Dennis R.

    1988-01-01

    Microgravity offers new ways of handling fluids, gases, and growing mammalian cells in efficient suspension cultures. In 1976 bioreactor engineers designed a system using a cylindrical reactor vessel in which the cells and medium are slowly mixed. The reaction chamber is interchangeable and can be used for several types of cell cultures. NASA has methodically developed unique suspension type cell and recovery apparatus culture systems for bioprocess technology experiments and production of biological products in microgravity. The first Space Bioreactor was designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small Bioreactor is being constructed for flight experiments in the Shuttle Middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption and control of low shear stress on cells.

  13. Optimizing of Culture Condition in Horizontal Rotating Bioreactor

    Institute of Scientific and Technical Information of China (English)

    Yan-Fang ZHANG; Huai-Qing CHEN; Hua HUANG

    2005-01-01

    @@ 1 Introduction Bioreactor is the most important equipment in tissue engineering. It can mimic the micro-environment of cell growth in vitro. At present, horizontal rotating bioreactor is the most advanced equipment for cell culture in the world.

  14. Analysis of Tubespins as a suitable scale-down model of bioreactors for high cell density CHO cell culture.

    Science.gov (United States)

    Gomez, Natalia; Ambhaikar, Malhar; Zhang, Li; Huang, Chung-Jr; Barkhordarian, Hedieh; Lull, Jonathan; Gutierrez, Chris

    2017-03-01

    High cell density (HCD) culture increases recombinant protein productivity via higher biomass. Compared to traditional fed-batch cultures, HCD is achieved by increased nutrient availability and removal of undesired metabolic components via regular medium replenishment. HCD process development is usually performed in instrumented lab-scale bioreactors (BR) that require time and labor for setup and operation. To potentially minimize resources and cost during HCD experiments, we evaluated a 2-week 50-mL Tubespin (TS) simulated HCD process where daily medium exchanges mimic the medium replacement rate in BR. To best assess performance differences, we cultured 13 different CHO cell lines in simulated HCD as satellites from simultaneous BR, and compared growth, metabolism, productivity and product quality. Overall, viability, cell-specific productivity and metabolism in TS were comparable to BR, but TS cell growth and final titer were lower by 25 and 15% in average, respectively. Peak viable cell densities were lower in TS than BR as a potential consequence of lower pH, different medium exchange strategy and dissolved oxygen limitations. Product quality attributes highly dependent on intrinsic molecule or cell line characteristics (e.g., galactosylation, afucosylation, aggregation) were comparable in both scales. However, product quality attributes that can change extracellularly as a function of incubation time (e.g., deamidation, C-terminal lysine, fragmentation) were in general lower in TS because of shorter residence time than HCD BR. Our characterization results and two case studies show that TS-simulated HCD cultures can be effectively used as a simple scale-down model for relative comparisons among cell lines for growth or productivity (e.g., clone screening), and for investigating effects on protein galactosylation. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:490-499, 2017. © 2017 American Institute of Chemical Engineers.

  15. Two new disposable bioreactors for plant cell culture: The wave and undertow bioreactor and the slug bubble bioreactor.

    Science.gov (United States)

    Terrier, Bénédicte; Courtois, Didier; Hénault, Nicolas; Cuvier, Arnaud; Bastin, Maryse; Aknin, Aziz; Dubreuil, Julien; Pétiard, Vincent

    2007-04-01

    The present article describes two novel flexible plastic-based disposable bioreactors. The first one, the WU bioreactor, is based on the principle of a wave and undertow mechanism that provides agitation while offering convenient mixing and aeration to the plant cell culture contained within the bioreactor. The second one is a high aspect ratio bubble column bioreactor, where agitation and aeration are achieved through the intermittent generation of large diameter bubbles, "Taylor-like" or "slug bubbles" (SB bioreactor). It allows an easy volume increase from a few liters to larger volumes up to several hundred liters with the use of multiple units. The cultivation of tobacco and soya cells producing isoflavones is described up to 70 and 100 L working volume for the SB bioreactor and WU bioreactor, respectively. The bioreactors being disposable and pre-sterilized before use, cleaning, sterilization, and maintenance operations are strongly reduced or eliminated. Both bioreactors represent efficient and low cost cell culture systems, applicable to various cell cultures at small and medium scale, complementary to traditional stainless-steel bioreactors.

  16. 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-b...

  17. A simple eccentric stirred tank mini-bioreactor: mixing characterization and mammalian cell culture experiments.

    Science.gov (United States)

    Bulnes-Abundis, David; Carrillo-Cocom, Leydi M; Aráiz-Hernández, Diana; García-Ulloa, Alfonso; Granados-Pastor, Marisa; Sánchez-Arreola, Pamela B; Murugappan, Gayathree; Alvarez, Mario M

    2013-04-01

    In industrial practice, stirred tank bioreactors are the most common mammalian cell culture platform. However, research and screening protocols at the laboratory scale (i.e., 5-100 mL) rely primarily on Petri dishes, culture bottles, or Erlenmeyer flasks. There is a clear need for simple-easy to assemble, easy to use, easy to clean-cell culture mini-bioreactors for lab-scale and/or screening applications. Here, we study the mixing performance and culture adequacy of a 30 mL eccentric stirred tank mini-bioreactor. A detailed mixing characterization of the proposed bioreactor is presented. Laser induced fluorescence (LIF) experiments and computational fluid dynamics (CFD) computations are used to identify the operational conditions required for adequate mixing. Mammalian cell culture experiments were conducted with two different cell models. The specific growth rate and the maximum cell density of Chinese hamster ovary (CHO) cell cultures grown in the mini-bioreactor were comparable to those observed for 6-well culture plates, Erlenmeyer flasks, and 1 L fully instrumented bioreactors. Human hematopoietic stem cells were successfully expanded tenfold in suspension conditions using the eccentric mini-bioreactor system. Our results demonstrate good mixing performance and suggest the practicality and adequacy of the proposed mini-bioreactor.

  18. Design and Performance of an Automated Bioreactor for Cell Culture Experiments in a Microgravity Environment

    Science.gov (United States)

    Kim, Youn-Kyu; Park, Seul-Hyun; Lee, Joo-Hee; Choi, Gi-Hyuk

    2015-03-01

    In this paper, we describe the development of a bioreactor for a cell-culture experiment on the International Space Station (ISS). The bioreactor is an experimental device for culturing mouse muscle cells in a microgravity environment. The purpose of the experiment was to assess the impact of microgravity on the muscles to address the possibility of longterm human residence in space. After investigation of previously developed bioreactors, and analysis of the requirements for microgravity cell culture experiments, a bioreactor design is herein proposed that is able to automatically culture 32 samples simultaneously. This reactor design is capable of automatic control of temperature, humidity, and culture-medium injection rate; and satisfies the interface requirements of the ISS. Since bioreactors are vulnerable to cell contamination, the medium-circulation modules were designed to be a completely replaceable, in order to reuse the bioreactor after each experiment. The bioreactor control system is designed to circulate culture media to 32 culture chambers at a maximum speed of 1 ml/min, to maintain the temperature of the reactor at 36°C, and to keep the relative humidity of the reactor above 70%. Because bubbles in the culture media negatively affect cell culture, a de-bubbler unit was provided to eliminate such bubbles. A working model of the reactor was built according to the new design, to verify its performance, and was used to perform a cell culture experiment that confirmed the feasibility of this device.

  19. A human hepatoma cell line FLC4 cultured on the radial flow bioreactor as a model for human hepatocytes

    Institute of Scientific and Technical Information of China (English)

    LiYW; BabuE

    2002-01-01

    Hepatocytes play central roles in the metabolism and excretion of drugs and xenobiovics.For this purpose,hepatocytes were endowed with high levels of enzyme activity for the phase I and phase Ⅱ metabolism as well as high levels of transmembrane transport activity which enables the entrance and the exit of drugs and xenobiotics and their metabolites through the plasma membrane of the hepatocytes.They include the transporters in the canalicular and sinusoidal membrane.Although a lot of cell lines were established from hepatoma cells or normal hepatocytes,none of them are fully satisfactory in the expression of the enzymes and transportens.We have established and characterized a hepatoma cell line designated FLC4 and found that this cell line exhibits properties quite similar to those of the normal hepatocytes in the light of enzymes and transporters for drug metabolism and transkport when they are cultured on the radial flow bioreactors.Using FLC4 cells cultured on the radial flow bioreactors,we are developing in vitro systems to evaluate the interaction of drugs with liver transporters and drug-drug interaction through the hepa tocyte transporters.

  20. A Versatile Bioreactor for Dynamic Suspension Cell Culture. Application to the Culture of Cancer Cell Spheroids.

    Science.gov (United States)

    Massai, Diana; Isu, Giuseppe; Madeddu, Denise; Cerino, Giulia; Falco, Angela; Frati, Caterina; Gallo, Diego; Deriu, Marco A; Falvo D'Urso Labate, Giuseppe; Quaini, Federico; Audenino, Alberto; Morbiducci, Umberto

    2016-01-01

    A versatile bioreactor suitable for dynamic suspension cell culture under tunable shear stress conditions has been developed and preliminarily tested culturing cancer cell spheroids. By adopting simple technological solutions and avoiding rotating components, the bioreactor exploits the laminar hydrodynamics establishing within the culture chamber enabling dynamic cell suspension in an environment favourable to mass transport, under a wide range of tunable shear stress conditions. The design phase of the device has been supported by multiphysics modelling and has provided a comprehensive analysis of the operating principles of the bioreactor. Moreover, an explanatory example is herein presented with multiphysics simulations used to set the proper bioreactor operating conditions for preliminary in vitro biological tests on a human lung carcinoma cell line. The biological results demonstrate that the ultralow shear dynamic suspension provided by the device is beneficial for culturing cancer cell spheroids. In comparison to the static suspension control, dynamic cell suspension preserves morphological features, promotes intercellular connection, increases spheroid size (2.4-fold increase) and number of cycling cells (1.58-fold increase), and reduces double strand DNA damage (1.5-fold reduction). It is envisioned that the versatility of this bioreactor could allow investigation and expansion of different cell types in the future.

  1. 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.

  2. The nitric oxide donor S-nitrosoglutathione reduces apoptotic primary liver cell loss in a three-dimensional perfusion bioreactor culture model developed for liver support.

    Science.gov (United States)

    Prince, Jose M; Vodovotz, Yoram; Baun, Matthew J; Monga, Satdarshan Pal; Billiar, Timothy R; Gerlach, Jörg C

    2010-03-01

    Artificial extracorporeal support for hepatic failure has met with limited clinical success. In hepatocytes, nitric oxide (NO) functions as an antiapoptotic modulator in response to a variety of stresses. We hypothesized that NO administration would yield improved viability and hepatocellular restructuring in a four-compartment, hollow fiber-based bioreactor with integral oxygenation for dynamic three-dimensional perfusion of hepatic cells in bioartificial liver support systems. Isolated adult rat liver cells were placed in culture medium alone (control) or medium supplemented with various concentrations of an NO donor (S-nitrosoglutathione [GSNO]) in the bioreactors. Media samples were obtained from the cell perfusion circuit to monitor cellular response. After 24 and 72 h, histology biopsies were taken to investigate spontaneous restructuring of the cells. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to quantify apoptotic nuclei. Control bioreactors exhibited 47.9 +/- 2.9% (mean +/- standard error of the mean) apoptotic nuclei. In contrast, NO-treated bioreactors exhibited a biphasic response. Fewer apoptotic nuclei were seen in the 200 and 500 microM GSNO groups (14.4 +/- 0.4%). No effect was observed in the 10 microM GSNO group (47.3%), and increased TUNEL staining was observed in the 1000 microM GSNO group (82.6%). Media lactate dehydrogenase levels were lower in bioreactor groups treated with 200 or 500 microM GSNO (310 +/- 38 IU/L) compared with the control group (919 +/- 188 IU/L; p bioreactors at 24 h vs. 110 +/- 13 in controls; p = 0.851). Histologically, all of the bioreactor groups exhibited liver cell aggregates with some attached to the bioreactor capillaries. Increased numbers of cells in the aggregates and superior spontaneous restructuring of the cells were seen at 24 and 72 h in the bioreactor groups treated with either 200 or 500 microM GSNO compared with the control groups. Addition of an NO donor

  3. Cell culture experiments planned for the space bioreactor

    Science.gov (United States)

    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.

  4. Hydrofocusing Bioreactor for Three-Dimensional Cell Culture

    Science.gov (United States)

    Gonda, Steve R.; Spaulding, Glenn F.; Tsao, Yow-Min D.; Flechsig, Scott; Jones, Leslie; Soehnge, Holly

    2003-01-01

    The hydrodynamic focusing bioreactor (HFB) is a bioreactor system designed for three-dimensional cell culture and tissue-engineering investigations on orbiting spacecraft and in laboratories on Earth. The HFB offers a unique hydrofocusing capability that enables the creation of a low-shear culture environment simultaneously with the "herding" of suspended cells, tissue assemblies, and air bubbles. Under development for use in the Biotechnology Facility on the International Space Station, the HFB has successfully grown large three-dimensional, tissuelike assemblies from anchorage-dependent cells and grown suspension hybridoma cells to high densities. The HFB, based on the principle of hydrodynamic focusing, provides the capability to control the movement of air bubbles and removes them from the bioreactor without degrading the low-shear culture environment or the suspended three-dimensional tissue assemblies. The HFB also provides unparalleled control over the locations of cells and tissues within its bioreactor vessel during operation and sampling.

  5. Bioreactors addressing diabetes mellitus.

    Science.gov (United States)

    Minteer, Danielle M; Gerlach, Jorg C; Marra, Kacey G

    2014-11-01

    The concept of bioreactors in biochemical engineering is a well-established process; however, the idea of applying bioreactor technology to biomedical and tissue engineering issues is relatively novel and has been rapidly accepted as a culture model. Tissue engineers have developed and adapted various types of bioreactors in which to culture many different cell types and therapies addressing several diseases, including diabetes mellitus types 1 and 2. With a rising world of bioreactor development and an ever increasing diagnosis rate of diabetes, this review aims to highlight bioreactor history and emerging bioreactor technologies used for diabetes-related cell culture and therapies.

  6. The culture of human embryonic stem cells in microchannel perfusion bioreactors

    Science.gov (United States)

    Korin, Natanel; Bransky, Avishay; Dinnar, Uri; Levenberg, Shulamit

    2007-12-01

    The culture of human Embryonic Stem (ES) cells in microchannel bioreactors can be highly beneficial for ES cell biology studies and ES tissue engineering applications. In the present study we examine the use of Human Foreskin Fibroblasts (HFF) cells as feeder cells for human ES culture in a microchannel perfusion bioreactor. PDMS microchannels (depth:130 micron) were fabricated using conventional soft-lithography techniques. The channels were sterilized, coated with a human fibronectin solution and seeded with cells. Following a period of static incubation, culture medium was perfused through the channels at various flow rates and cell growth was monitored throughout the culture process. Mass transport and fluid mechanics models were used to evaluate the culture conditions (shear stress, oxygen levels within the micro-bioreactor as a function of the medium flow rate. The conditions for successful long-term culture (>7 days) of HFF under flow were established. Experiments with human embryonic stem cells cultured in microchannels show that the conditions essential to co-culture human ES cell on HFF cells under perfusion differ from the conditions necessary for HFF cell culture. Human ES cells were found to be highly sensitive to flow and culture conditions and did not grow under flow rates which were suitable for HFF long-term culture. Successful culture of undifferentiated human ES cell colonies in a perfusion micro-bioreactor is a basic step towards utilizing microfluidic techniques to explore stem cell biology.

  7. Advanced microscale bioreactor system: a representative scale-down model for bench-top bioreactors.

    Science.gov (United States)

    Hsu, Wei-Ting; Aulakh, Rigzen P S; Traul, Donald L; Yuk, Inn H

    2012-12-01

    In recent years, several automated scale-down bioreactor systems have been developed to increase efficiency in cell culture process development. ambr™ is an automated workstation that provides individual monitoring and control of culture dissolved oxygen and pH in single-use, stirred-tank bioreactors at a working volume of 10-15 mL. To evaluate the ambr™ system, we compared the performance of four recombinant Chinese hamster ovary cell lines in a fed-batch process in parallel ambr™, 2-L bench-top bioreactors, and shake flasks. Cultures in ambr™ matched 2-L bioreactors in controlling the environment (temperature, dissolved oxygen, and pH) and in culture performance (growth, viability, glucose, lactate, Na(+), osmolality, titer, and product quality). However, cultures in shake flasks did not show comparable performance to the ambr™ and 2-L bioreactors.

  8. 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.

  9. Over-pressurized bioreactors: application to microbial cell cultures.

    Science.gov (United States)

    Lopes, Marlene; Belo, Isabel; Mota, Manuel

    2014-01-01

    In industrial biotechnology, microbial cultures are exposed to different local pressures inside bioreactors. Depending on the microbial species and strains, the increased pressure may have detrimental or beneficial effects on cellular growth and product formation. In this review, the effects of increased air pressure on various microbial cultures growing in bioreactors under moderate total pressure conditions (maximum, 15 bar) will be discussed. Recent data illustrating the diversity of increased air pressure effects at different levels in microbial cells cultivation will be presented, with particular attention to the effects of oxygen and carbon dioxide partial pressures on cellular growth and product formation, and the concomitant effect of oxygen pressure on antioxidant cellular defense mechanisms.

  10. Three-dimensional perfusion bioreactor culture supports differentiation of human fetal liver cells.

    Science.gov (United States)

    Schmelzer, Eva; Triolo, Fabio; Turner, Morris E; Thompson, Robert L; Zeilinger, Katrin; Reid, Lola M; Gridelli, Bruno; Gerlach, Jörg C

    2010-06-01

    The ability of human fetal liver cells to survive, expand, and form functional tissue in vitro is of high interest for the development of bioartificial extracorporeal liver support systems, liver cell transplantation therapies, and pharmacologic models. Conventional static two-dimensional culture models seem to be inadequate tools. We focus on dynamic three-dimensional perfusion technologies and developed a scaled-down bioreactor, providing decentralized mass exchange with integral oxygenation. Human fetal liver cells were embedded in a hyaluronan hydrogel within the capillary system to mimic an in vivo matrix and perfusion environment. Metabolic performance was monitored daily, including glucose consumption, lactate dehydrogenase activity, and secretion of alpha-fetoprotein and albumin. At culture termination cells were analyzed for proliferation and liver-specific lineage-dependent cytochrome P450 (CYP3A4/3A7) gene expression. Occurrence of hepatic differentiation in bioreactor cultures was demonstrated by a strong increase in CYP3A4/3A7 gene expression ratio, lower alpha-fetoprotein, and higher albumin secretion than in conventional Petri dish controls. Cells in bioreactors formed three-dimensional structures. Viability of cells was higher in bioreactors than in control cultures. In conclusion, the culture model implementing three-dimensionality, constant perfusion, and integral oxygenation in combination with a hyaluronan hydrogel provides superior conditions for liver cell survival and differentiation compared to conventional culture.

  11. Microfluidic bioreactors for culture of non-adherent cells

    DEFF Research Database (Denmark)

    Shah, Pranjul Jaykumar; Vedarethinam, Indumathi; Kwasny, Dorota

    2011-01-01

    Microfluidic bioreactors (μBR) are becoming increasingly popular for cell culture, sample preparation and analysis in case of routine genetic and clinical diagnostics. We present a novel μBR for non-adherent cells designed to mimic in vivo perfusion of cells based on diffusion of media through...

  12. Three-Dimensional Modelling inside a Differential Pressure Laminar Flow Bioreactor Filled with Porous Media

    Directory of Open Access Journals (Sweden)

    Birgit Weyand

    2015-01-01

    Full Text Available A three-dimensional computational fluid dynamics- (CFD- model based on a differential pressure laminar flow bioreactor prototype was developed to further examine performance under changing culture conditions. Cell growth inside scaffolds was simulated by decreasing intrinsic permeability values and led to pressure build-up in the upper culture chamber. Pressure release by an integrated bypass system allowed continuation of culture. The specific shape of the bioreactor culture vessel supported a homogenous flow profile and mass flux at the scaffold level at various scaffold permeabilities. Experimental data showed an increase in oxygen concentration measured inside a collagen scaffold seeded with human mesenchymal stem cells when cultured in the perfusion bioreactor after 24 h compared to static culture in a Petri dish (dynamic: 11% O2 versus static: 3% O2. Computational fluid simulation can support design of bioreactor systems for tissue engineering application.

  13. Long-term three-dimensional perfusion culture of human adult bone marrow mononuclear cells in bioreactors.

    Science.gov (United States)

    Schmelzer, Eva; Finoli, Anthony; Nettleship, Ian; Gerlach, Jörg C

    2015-04-01

    The construction and long-term maintenance of three-dimensional in vitro bone marrow models is of great interest but still quite challenging. Here we describe the use of a multi-compartment hollow-fiber membrane based three-dimensional perfusion bioreactor for long-term culture of whole human bone marrow mononuclear cells. We also investigated bioreactors with incorporated open-porous foamed hydroxyapatite scaffolds, mimicking the in vivo bone matrix. Cells in bioreactors with and without scaffolds were cultured to 6 weeks and compared to Petri dish controls. Cells were analyzed for gene expression, surface markers by flow cytometry, metabolic activity, hematopoietic potential, viability, and attachment by immunocytochemistry. Cells in bioreactors were metabolic active during long-term culture. The percentages of hematopoietic stem cell and mature endothelial cell fractions were maintained in bioreactors. The expression of most of the analyzed genes stabilized and increased after long-term culture of 6 weeks. Compared to Petri dish culture controls, bioreactor perfusion culture improved in both the short and long-term, the colony formation unit capacity of hematopoietic progenitors. Cells attached to the ample surface area provided by hydroxyapatite scaffolds. The implementation of a hydroxyapatite scaffold did not influence colony formation capacity, percentages of cell type specific fractions, gene expression, cell viability or metabolic turnover when compared to control cells cultured in bioreactors without scaffolds. In conclusion, three-dimensional perfusion bioreactor culture enables long-term maintenance of primary human bone marrow cells, with hydroxyapatite scaffolds providing an in vivo-like scaffold for three-dimensional culture. © 2015 Wiley Periodicals, Inc.

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

    Science.gov (United States)

    Rowland-Jones, Ruth C; van den Berg, Frans; Racher, Andrew J; Martin, Elaine B; Jaques, Colin

    2017-03-01

    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 number of offline samples that can be taken compared to those taken for monitoring cultures in large-scale bioreactors. The small volume of miniature bioreactor cultures (15 mL) is incompatible with the large sample volume (600 µL) required for bioanalysers routinely used. Spectroscopy technologies may be used to resolve this limitation. The purpose of this study was to compare the use of NIR, Raman, and 2D-fluorescence to measure multiple analytes simultaneously in volumes suitable for daily monitoring of a miniature bioreactor system. A novel design-of-experiment approach is described that utilizes previously analyzed cell culture supernatant to assess metabolite concentrations under various conditions while providing optimal coverage of the desired design space. Multivariate data analysis techniques were used to develop predictive models. Model performance was compared to determine which technology is more suitable for this application. 2D-fluorescence could more accurately measure ammonium concentration (RMSECV 0.031 g L(-1) ) than Raman and NIR. Raman spectroscopy, however, was more robust at measuring lactate and glucose concentrations (RMSECV 1.11 and 0.92 g L(-1) , respectively) than the other two techniques. The findings suggest that Raman spectroscopy is more suited for this application than NIR and 2D-fluorescence. The implementation of Raman spectroscopy increases at-line measuring capabilities, enabling daily monitoring of key cell culture components within miniature bioreactor cultures. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:337-346, 2017. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on

  15. 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.

  16. 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.

  17. In vivo bone regeneration using tubular perfusion system bioreactor cultured nanofibrous scaffolds

    NARCIS (Netherlands)

    Yeatts, A.B.; Both, S.K.; Yang, W.; Alghamdi, H.S.A.; Yang, F.; Fisher, J.P.; Jansen, J.A.

    2014-01-01

    The use of bioreactors for the in vitro culture of constructs for bone tissue engineering has become prevalent as these systems may improve the growth and differentiation of a cultured cell population. Here we utilize a tubular perfusion system (TPS) bioreactor for the in vitro culture of human

  18. Long-term immunologically competent human peripheral lymphoid tissue cultures in a 3D bioreactor

    Science.gov (United States)

    Kuzin, Igor; Sun, Hongliang; Moshkani, Safiekhatoon; Feng, Changyong; Mantalaris, Athanasios; Wu, JH David; Bottaro, Andrea

    2011-01-01

    Peripheral lymphoid organs (PLOs), the primary sites of development of adaptive immune responses, display a complex structural organization reflecting separation of cellular subsets (e.g. T and B lymphocytes) and functional compartments which is critical for immune function. The generation of in vitro culture systems capable of recapitulating salient features of PLOs for experimental, biotechnological and clinical applications would be highly desirable, but has been hampered so far by the complexity of these systems. We have previously developed a three-dimensional bioreactor system for long-term, functional culture of human bone marrow cells on macroporous microspheres in a packed-bed bioreactor with frequent medium change. Here we adapt the same system for culture of human primary cells from PLOs (tonsil) in the absence of specific exogenous growth factors or activators. Cells in this system displayed higher viability over several weeks, and maintain population diversity and cell surface markers largely comparable to primary cells. Light microscopy showed cells organizing in large diverse clusters within the scaffold pores and presence of B cell-enriched areas. Strikingly, these cultures generated a significant number of antibody-producing B cells when challenged with a panel of diverse antigens, as expected from a lymphoid tissue. Thus the three-dimensional tonsil bioreactor culture system may serve as a useful model of PLOs by recapitulating their structural organization and function ex vivo. PMID:21309085

  19. Microliter-bioreactor array with buoyancy-driven stirring for human hematopoietic stem cell culture

    OpenAIRE

    Luni, Camilla; Feldman, Hope C.; Pozzobon, Michela; De Coppi, Paolo; Meinhart, Carl D.; Elvassore, Nicola

    2010-01-01

    This work presents the development of an array of bioreactors where finely controlled stirring is provided at the microliter scale (100–300 μl). The microliter-bioreactor array is useful for performing protocol optimization in up to 96 parallel experiments of hematopoietic stem cell (HSC) cultures. Exploring a wide range of experimental conditions at the microliter scale minimizes cost and labor. Once the cell culture protocol is optimized, it can be applied to large-scale bioreactors for ste...

  20. The bioreactor: a powerful tool for large-scale culture of animal cells.

    Science.gov (United States)

    Wang, Dianliang; Liu, Wanshun; Han, Baoqin; Xu, Ruian

    2005-10-01

    Bioreactors play a key role in the field of biologics, where they are used for the production of recombinant therapeutic proteins by large-scale cultivation of animal cells. There are several types of bioreactors, including stirred-tank, airlift, hollow-fiber, and Rotary Cell Culture System (RCCS) designs. The stirred-tank bioreactor is one of the most commonly used types, and is used both for industrial applications and laboratory research. The RCCS, invented by NASA, is increasingly used in the area of tissue engineering for medical purposes. Important improvements have been made in the design of traditional bioreactors, and new types of bioreactor are also being developed such as Couette-Taylor bioreactor, multifunctional-membrane bioreactor, and shaking bioreactor. Work is also progressing on techniques to improve the performance of bioreactors, including perfusion culture, the use of microcarriers, and methods of suppressing apoptosis and of monitoring cell growth in real time. Given the demand for the production by animal cells for use in the growing number of clinical applications, further advances in bioreactor technology can be expected during the next few years. Two main goals will be pursued: firstly, to increase output by high density cultivation of animal cells to produce high value protein pharmaceutics or viral vectors for clinical gene therapy; and secondly, to create a three-dimension space similar to that of an in vivo environment to regenerate tissue or organ and to reproduce valuable cells that are hard to culture in the traditional culture system.

  1. Cell Cycle Progression of Human Cells Cultured in Rotating Bioreactor

    Science.gov (United States)

    Parks, Kelsey

    2009-01-01

    Space flight has been shown to alter the astronauts immune systems. Because immune performance is complex and reflects the influence of multiple organ systems within the host, scientists sought to understand the potential impact of microgravity alone on the cellular mechanisms critical to immunity. Lymphocytes and their differentiated immature form, lymphoblasts, play an important and integral role in the body's defense system. T cells, one of the three major types of lymphocytes, play a central role in cell-mediated immunity. They can be distinguished from other lymphocyte types, such as B cells and natural killer cells by the presence of a special receptor on their cell surface called T cell receptors. Reported studies have shown that spaceflight can affect the expression of cell surface markers. Cell surface markers play an important role in the ability of cells to interact and to pass signals between different cells of the same phenotype and cells of different phenotypes. Recent evidence suggests that cell-cycle regulators are essential for T-cell function. To trigger an effective immune response, lymphocytes must proliferate. The objective of this project is to investigate the changes in growth of human cells cultured in rotating bioreactors and to measure the growth rate and the cell cycle distribution for different human cell types. Human lymphocytes and lymphoblasts will be cultured in a bioreactor to simulate aspects of microgravity. The bioreactor is a cylindrical culture vessel that incorporates the aspects of clinostatic rotation of a solid fluid body around a horizontal axis at a constant speed, and compensates gravity by rotation and places cells within the fluid body into a sustained free-fall. Cell cycle progression and cell proliferation of the lymphocytes will be measured for a number of days. In addition, RNA from the cells will be isolated for expression of genes related in cell cycle regulations.

  2. Cell Cycle Progression of Human Cells Cultured in Rotating Bioreactor

    Science.gov (United States)

    Parks, Kelsey

    2009-01-01

    Space flight has been shown to alter the astronauts immune systems. Because immune performance is complex and reflects the influence of multiple organ systems within the host, scientists sought to understand the potential impact of microgravity alone on the cellular mechanisms critical to immunity. Lymphocytes and their differentiated immature form, lymphoblasts, play an important and integral role in the body's defense system. T cells, one of the three major types of lymphocytes, play a central role in cell-mediated immunity. They can be distinguished from other lymphocyte types, such as B cells and natural killer cells by the presence of a special receptor on their cell surface called T cell receptors. Reported studies have shown that spaceflight can affect the expression of cell surface markers. Cell surface markers play an important role in the ability of cells to interact and to pass signals between different cells of the same phenotype and cells of different phenotypes. Recent evidence suggests that cell-cycle regulators are essential for T-cell function. To trigger an effective immune response, lymphocytes must proliferate. The objective of this project is to investigate the changes in growth of human cells cultured in rotating bioreactors and to measure the growth rate and the cell cycle distribution for different human cell types. Human lymphocytes and lymphoblasts will be cultured in a bioreactor to simulate aspects of microgravity. The bioreactor is a cylindrical culture vessel that incorporates the aspects of clinostatic rotation of a solid fluid body around a horizontal axis at a constant speed, and compensates gravity by rotation and places cells within the fluid body into a sustained free-fall. Cell cycle progression and cell proliferation of the lymphocytes will be measured for a number of days. In addition, RNA from the cells will be isolated for expression of genes related in cell cycle regulations.

  3. Hepatocyte function within a stacked double sandwich culture plate cylindrical bioreactor for bioartificial liver system.

    Science.gov (United States)

    Xia, Lei; Arooz, Talha; Zhang, Shufang; Tuo, Xiaoye; Xiao, Guangfa; Susanto, Thomas Adi Kurnia; Sundararajan, Janani; Cheng, Tianming; Kang, Yuzhan; Poh, Hee Joo; Leo, Hwa Liang; Yu, Hanry

    2012-11-01

    Bioartificial liver (BAL) system is promising as an alternative treatment for liver failure. We have developed a bioreactor with stacked sandwich culture plates for the application of BAL. This bioreactor design addresses some of the persistent problems in flat-bed bioreactors through increasing cell packing capacity, eliminating dead flow, regulating shear stress, and facilitating the scalability of the bioreactor unit. The bioreactor contained a stack of twelve double-sandwich-culture plates, allowing 100 million hepatocytes to be housed in a single cylindrical bioreactor unit (7 cm of height and 5.5 cm of inner diameter). The serial flow perfusion through the bioreactor increased cell-fluid contact area for effective mass exchange. With the optimal perfusion flow rate, shear stress was minimized to achieve high and uniform cell viabilities across different plates in the bioreactor. Our results demonstrated that hepatocytes cultured in the bioreactor could re-establish cell polarity and maintain liver-specific functions (e.g. albumin and urea synthesis, phase I&II metabolism functions) for seven days. The single bioreactor unit can be readily scaled up to house adequate number of functional hepatocytes for BAL development.

  4. A comparison of bioreactors for culture of fetal mesenchymal stem cells for bone tissue engineering.

    Science.gov (United States)

    Zhang, Zhi-Yong; Teoh, Swee Hin; Teo, Erin Yiling; Khoon Chong, Mark Seow; Shin, Chong Woon; Tien, Foo Toon; Choolani, Mahesh A; Chan, Jerry K Y

    2010-11-01

    Bioreactors provide a dynamic culture system for efficient exchange of nutrients and mechanical stimulus necessary for the generation of effective tissue engineered bone grafts (TEBG). We have shown that biaxial rotating (BXR) bioreactor-matured human fetal mesenchymal stem cell (hfMSC) mediated-TEBG can heal a rat critical sized femoral defect. However, it is not known whether optimal bioreactors exist for bone TE (BTE) applications. We systematically compared this BXR bioreactor with three most commonly used systems: Spinner Flask (SF), Perfusion and Rotating Wall Vessel (RWV) bioreactors, for their application in BTE. The BXR bioreactor achieved higher levels of cellularity and confluence (1.4-2.5x, p bioreactors operating in optimal settings. BXR bioreactor-treated scaffolds experienced earlier and more robust osteogenic differentiation on von Kossa staining, ALP induction (1.2-1.6×, p bioreactor-treated grafts, but not with the other three. BXR bioreactor enabled superior cellular proliferation, spatial distribution and osteogenic induction of hfMSC over other commonly used bioreactors. In addition, we developed and validated a non-invasive quantitative micro CT-based technique for analyzing neo-tissue formation and its spatial distribution within scaffolds.

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

    Science.gov (United States)

    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.

  6. Novel Bioreactors to Study Forces on Bronchial Epithelial Cultures

    Science.gov (United States)

    Carpenter, Jerome; Millard, Mike; Cozon, Matthew; Superfine, Richard

    2008-10-01

    Studying cells in a physiologically relevant environment is an important tool in understanding cell signaling and gene expression. Human bronchial epithelial cells (HBECs) are responsible for mucociliary clearance, which removes pathogens from the air we breathe. Recreating the in vivo conditions of HBECs is difficult; they are polarized and undergo a variety of forces. Polarization is required for organ-specific systems such as cilia motility and mucus regulation. We achieve polarization by growing cells on an electrospun nanoporous scaffold which we attach to a silastic annulus. Using this geometry we apply vacuum to the annulus and stretch the cells. This bioreactor allows us to study polarized HBECs as they experience cyclic strain similar to breathing. We've grown polarized cultures on the scaffold and are evaluating the scaffold's mechanical properties. In a second bioreactor, we place the scaffold into a microfluidics channel to study the affect of shear stress on polarized cells. We also reproduce the branching structure found in the lungs to investigate the regulation of mucus as it ascends the airway tree.

  7. Modelling across bioreactor scales: methods, challenges and limitations

    DEFF Research Database (Denmark)

    Gernaey, Krist

    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...... 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...... are the limitations of different types of mod - els? This paper will provide examples of models that have been published in the literature for use across bioreactor scales, including computational fluid dynamics (CFD) and population balance models. Furthermore, the importance of good modeling practice...

  8. Teratoma formation of human embryonic stem cells in three-dimensional perfusion culture bioreactors.

    Science.gov (United States)

    Stachelscheid, H; Wulf-Goldenberg, A; Eckert, K; Jensen, J; Edsbagge, J; Björquist, P; Rivero, M; Strehl, R; Jozefczuk, J; Prigione, A; Adjaye, J; Urbaniak, T; Bussmann, P; Zeilinger, K; Gerlach, J C

    2013-09-01

    Teratoma formation in mice is today the most stringent test for pluripotency that is available for human pluripotent cells, as chimera formation and tetraploid complementation cannot be performed with human cells. The teratoma assay could also be applied for assessing the safety of human pluripotent cell-derived cell populations intended for therapeutic applications. In our study we examined the spontaneous differentiation behaviour of human embryonic stem cells (hESCs) in a perfused 3D multi-compartment bioreactor system and compared it with differentiation of hESCs and human induced pluripotent cells (hiPSCs) cultured in vitro as embryoid bodies and in vivo in an experimental mouse model of teratoma formation. Results from biochemical, histological/immunohistological and ultrastuctural analyses revealed that hESCs cultured in bioreactors formed tissue-like structures containing derivatives of all three germ layers. Comparison with embryoid bodies and the teratomas revealed a high degree of similarity of the tissues formed in the bioreactor to these in the teratomas at the histological as well as transcriptional level, as detected by comparative whole-genome RNA expression profiling. The 3D culture system represents a novel in vitro model that permits stable long-term cultivation, spontaneous multi-lineage differentiation and tissue formation of pluripotent cells that is comparable to in vivo differentiation. Such a model is of interest, e.g. for the development of novel cell differentiation strategies. In addition, the 3D in vitro model could be used for teratoma studies and pluripotency assays in a fully defined, controlled environment, alternatively to in vivo mouse models.

  9. Oxygen transport and stem cell aggregation in stirred-suspension bioreactor cultures.

    Directory of Open Access Journals (Sweden)

    Jincheng Wu

    Full Text Available Stirred-suspension bioreactors are a promising modality for large-scale culture of 3D aggregates of pluripotent stem cells and their progeny. Yet, cells within these clusters experience limitations in the transfer of factors and particularly O2 which is characterized by low solubility in aqueous media. Cultured stem cells under different O2 levels may exhibit significantly different proliferation, viability and differentiation potential. Here, a transient diffusion-reaction model was built encompassing the size distribution and ultrastructural characteristics of embryonic stem cell (ESC aggregates. The model was coupled to experimental data from bioreactor and static cultures for extracting the effective diffusivity and kinetics of consumption of O2 within mouse (mESC and human ESC (hESC clusters. Under agitation, mESC aggregates exhibited a higher maximum consumption rate than hESC aggregates. Moreover, the reaction-diffusion model was integrated with a population balance equation (PBE for the temporal distribution of ESC clusters changing due to aggregation and cell proliferation. Hypoxia was found to be negligible for ESCs with a smaller radius than 100 µm but became appreciable for aggregates larger than 300 µm. The integrated model not only captured the O2 profile both in the bioreactor bulk and inside ESC aggregates but also led to the calculation of the duration that fractions of cells experience a certain range of O2 concentrations. The approach described in this study can be employed for gaining a deeper understanding of the effects of O2 on the physiology of stem cells organized in 3D structures. Such frameworks can be extended to encompass the spatial and temporal availability of nutrients and differentiation factors and facilitate the design and control of relevant bioprocesses for the production of stem cell therapeutics.

  10. 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.

  11. [Research on ursolic acid production of Eriobotrya japonica cell suspension culture in WAVE bioreactor].

    Science.gov (United States)

    Li, Hui-hua; Yao, De-heng; Xu, Jian; Wang, Wei; Chang, Qiang; Su, Ming-hua

    2015-05-01

    Through scale-up cultivation of Eriobotrya japonica suspension cells using WAVE bioreactor, the cell growth and ursolic acid (UA) accumulation were studied. The comparison test was carried out in the flask and the reactor with cell dry weight (DW) and UA content as evaluation indexes. The culture medium, DW and UA content were compared in 1 L and 5 L working volumes of bioreactor. The orthogonal test with main actors of inoculation amount, speed and angle of rotation was developed to find the optimal combination, in 1 L working volume of bioreactor. DW of the cell growth and the UA content in bioreactor were higher than those of the shaker by 105.5% and 27.65% respectively. In bioreactor, the dynamic changes of elements in the fluid culture, the dry weight of the cell growth and the UA content in 1 L and 5 L working volumes were similar. Inoculation of 80 g, rotational speed of 26 r · min(-1), and angle of 6 ° was the optimal combination, and the cell biomass of 19.01 g · L(-1) and the UA content of 27.750 mg · g(-1) were achieved after 100 h cultivation in 1 L working volume of bioreactor. WAVE Bioreactor is more suitable than flasks for the E. japonica cell suspension culture, and culture parameters can be achieved from 1 L to 5 L amplification.

  12. 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

  13. In vivo bone regeneration using tubular perfusion system bioreactor cultured nanofibrous scaffolds.

    Science.gov (United States)

    Yeatts, Andrew B; Both, Sanne K; Yang, Wanxun; Alghamdi, Hamdan S; Yang, Fang; Fisher, John P; Jansen, John A

    2014-01-01

    The use of bioreactors for the in vitro culture of constructs for bone tissue engineering has become prevalent as these systems may improve the growth and differentiation of a cultured cell population. Here we utilize a tubular perfusion system (TPS) bioreactor for the in vitro culture of human mesenchymal stem cells (hMSCs) and implant the cultured constructs into rat femoral condyle defects. Using nanofibrous electrospun poly(lactic-co-glycolic acid)/poly(ε-caprolactone) scaffolds, hMSCs were cultured for 10 days in vitro in the TPS bioreactor with cellular and acellular scaffolds cultured statically for 10 days as a control. After 3 and 6 weeks of in vivo culture, explants were removed and subjected to histomorphometric analysis. Results indicated more rapid bone regeneration in defects implanted with bioreactor cultured scaffolds with a new bone area of 1.23 ± 0.35 mm(2) at 21 days compared to 0.99 ± 0.43 mm(2) and 0.50 ± 0.29 mm(2) in defects implanted with statically cultured scaffolds and acellular scaffolds, respectively. At the 21 day timepoint, statistical differences (pbioreactor to improve bone tissue regeneration and highlights the benefits of utilizing perfusion bioreactor systems to culture MSCs for bone tissue engineering.

  14. In Vivo Bone Regeneration Using Tubular Perfusion System Bioreactor Cultured Nanofibrous Scaffolds

    Science.gov (United States)

    Yeatts, Andrew B.; Both, Sanne K.; Yang, Wanxun; Alghamdi, Hamdan S.; Yang, Fang; Jansen, John A.

    2014-01-01

    The use of bioreactors for the in vitro culture of constructs for bone tissue engineering has become prevalent as these systems may improve the growth and differentiation of a cultured cell population. Here we utilize a tubular perfusion system (TPS) bioreactor for the in vitro culture of human mesenchymal stem cells (hMSCs) and implant the cultured constructs into rat femoral condyle defects. Using nanofibrous electrospun poly(lactic-co-glycolic acid)/poly(ɛ-caprolactone) scaffolds, hMSCs were cultured for 10 days in vitro in the TPS bioreactor with cellular and acellular scaffolds cultured statically for 10 days as a control. After 3 and 6 weeks of in vivo culture, explants were removed and subjected to histomorphometric analysis. Results indicated more rapid bone regeneration in defects implanted with bioreactor cultured scaffolds with a new bone area of 1.23±0.35 mm2 at 21 days compared to 0.99±0.43 mm2 and 0.50±0.29 mm2 in defects implanted with statically cultured scaffolds and acellular scaffolds, respectively. At the 21 day timepoint, statistical differences (pbioreactor to improve bone tissue regeneration and highlights the benefits of utilizing perfusion bioreactor systems to culture MSCs for bone tissue engineering. PMID:23865551

  15. 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.

  16. Development of thin-film photo-bioreactor and its application to outdoor culture of microalgae.

    Science.gov (United States)

    Yoo, Jae Jun; Choi, Seung Phill; Kim, Jaoon Y H; Chang, Won Seok; Sim, Sang Jun

    2013-06-01

    Photosynthetic microalgae have received much attention as a microbial source of diverse useful biomaterials through CO(2) fixation and various types of photo-bioreactors have been developed for efficient microalgal cultivation. Herein, we developed a novel thin-film photo-bioreactor, which was made of cast polypropylene film, considering outdoor mass cultivation. To develop optimal design of photo-bioreactor, we tested performance of three shapes of thin-film photo-bioreactors (flat, horizontal and vertical tubular shapes) and various parts in the bioreactor. Collectively, vertical tubular bioreactor with H/D ratio 6:1 and cylindrical stainless steel spargers showed the most outstanding performance. Furthermore, the photo-bioreactor was successfully applied to the cultivation of other microalgae such as Chlamydomonas reinhardtii and Chlorella vulgaris. The scalability of photo-bioreactor was confirmed by gradually increasing culture volume from 4 to 25 L and the biomass productivity of each reactor was quite consistent (0.05-0.07 g/L/day) during the cultivation of H. pluvialis under indoor and outdoor conditions. Especially, we also achieved dry cell weight of 4.64 g/L and astaxanthin yield of 218.16 mg/L through long-term cultivation (100 days) under outdoor condition in 15 L photo-bioreactor using Haematococcus pluvialis, which means that the astaxanthin yield from outdoor cultivation is equal or superior to that obtained from controlled indoor condition. Therefore, these results indicate that we can apply this approach to development of optimal photo-bioreactor for the large-scale culture of microalgae and production of useful biomaterials under outdoor condition.

  17. Continuous plant cell perfusion culture: bioreactor characterization and secreted enzyme production.

    Science.gov (United States)

    Su, Wei Wen; Arias, Renee

    2003-01-01

    Culture perfusion is widely practiced in mammalian cell processes to enhance secreted antibody production. Here, we report the development of an efficient continuous perfusion process for the cultivation of plant cell suspensions. The key to this process is a perfusion bioreactor that incorporates an annular settling zone into a stirred-tank bioreactor to achieve continuous cell/medium separation via gravitational sedimentation. From washout experiments, we found that under typical operating conditions (e.g., 200 rpm and 0.3 vvm) the liquid phase in the entire perfusion bioreactor was homogeneous despite the presence of the cylindrical baffle. Using secreted acid phosphatase (APase) produced in Anchusa officinalis cell culture as a model we have studied the perfusion cultures under complete or partial cell retention. The perfusion culture was operated under phosphate limitation to stimulate APase production. Successful operation of the perfusion process over four weeks has been achieved in this work. When A. officinalis cells were grown in the perfusion reactor and perfused at up to 0.4 vvd with complete cell retention, a cell dry weight exceeding 20 g/l could be achieved while secreted APase productivity leveled off at approximately 300 units/l/d. The culture became extremely dense with the maximum packed cell volume (PCV) surpassing 70%. In comparison, the maximum cell dry weight and overall secreted APase productivity in a typical batch culture were 10-12 g/l and 100-150 units/l/d, respectively. Operation of the perfusion culture under extremely high PCV for a prolonged period, however, led to declined oxygen uptake and reduced viability. Subsequently, cell removal via a bleed stream at up to 0.11 vvd was tested and shown to stabilize the culture at a PCV below 60%. With culture bleeding, both specific oxygen uptake rate and viability were shown to increase. This also led to a higher cell dry weight exceeding 25 g/l, and further improvement of secreted APase

  18. 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.

  19. 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

  20. Organ culture bioreactors--platforms to study human intervertebral disc degeneration and regenerative therapy.

    Science.gov (United States)

    Gantenbein, Benjamin; Illien-Jünger, Svenja; Chan, Samantha C W; Walser, Jochen; Haglund, Lisbet; Ferguson, Stephen J; Iatridis, James C; Grad, Sibylle

    2015-01-01

    In recent decades the application of bioreactors has revolutionized the concept of culturing tissues and organs that require mechanical loading. In intervertebral disc (IVD) research, collaborative efforts of biomedical engineering, biology and mechatronics have led to the innovation of new loading devices that can maintain viable IVD organ explants from large animals and human cadavers in precisely defined nutritional and mechanical environments over extended culture periods. Particularly in spine and IVD research, these organ culture models offer appealing alternatives, as large bipedal animal models with naturally occurring IVD degeneration and a genetic background similar to the human condition do not exist. Latest research has demonstrated important concepts including the potential of homing of mesenchymal stem cells to nutritionally or mechanically stressed IVDs, and the regenerative potential of "smart" biomaterials for nucleus pulposus or annulus fibrosus repair. In this review, we summarize the current knowledge about cell therapy, injection of cytokines and short peptides to rescue the degenerating IVD. We further stress that most bioreactor systems simplify the real in vivo conditions providing a useful proof of concept. Limitations are that certain aspects of the immune host response and pain assessments cannot be addressed with ex vivo systems. Coccygeal animal disc models are commonly used because of their availability and similarity to human IVDs. Although in vitro loading environments are not identical to the human in vivo situation, 3D ex vivo organ culture models of large animal coccygeal and human lumbar IVDs should be seen as valid alternatives for screening and feasibility testing to augment existing small animal, large animal, and human clinical trial experiments.

  1. NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    Bioreactor Demonstration System (BDS) comprises an electronics module, a gas supply module, and the incubator module housing the rotating wall vessel and its support systems. Nutrient media are pumped through an oxygenator and the culture vessel. The shell rotates at 0.5 rpm while the irner filter typically rotates at 11.5 rpm to produce a gentle flow that ensures removal of waste products as fresh media are infused. Periodically, some spent media are pumped into a waste bag and replaced by fresh media. When the waste bag is filled, an astronaut drains the waste bag and refills the supply bag through ports on the face of the incubator. Pinch valves and a perfusion pump ensure that no media are exposed to moving parts. An Experiment Control Computer controls the Bioreactor, records conditions, and alerts the crew when problems occur. The crew operates the system through a laptop computer displaying graphics designed for easy crew training and operation. 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. See No. 0101825 for a version with major elements labeled, and No. 0103180 for an operational schematic. 0101816

  2. NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    Bioreactor Demonstration System (BDS) comprises an electronics module, a gas supply module, and the incubator module housing the rotating wall vessel and its support systems. Nutrient media are pumped through an oxygenator and the culture vessel. The shell rotates at 0.5 rpm while the irner filter typically rotates at 11.5 rpm to produce a gentle flow that ensures removal of waste products as fresh media are infused. Periodically, some spent media are pumped into a waste bag and replaced by fresh media. When the waste bag is filled, an astronaut drains the waste bag and refills the supply bag through ports on the face of the incubator. Pinch valves and a perfusion pump ensure that no media are exposed to moving parts. An Experiment Control Computer controls the Bioreactor, records conditions, and alerts the crew when problems occur. The crew operates the system through a laptop computer displaying graphics designed for easy crew training and operation. 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. See No. 0101825 for a version with major elements labeled, and No. 0103180 for an operational schematic. 0101816

  3. NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    Biotechnology Specimen Temperature Controller (BSTC) will cultivate cells until their turn in the bioreactor; it can also be used in culturing experiments that do not require the bioreactor. The BSTC comprises four incubation/refrigeration chambers individually set at 4 to 50 deg. C (near-freezing to above body temperature). Each chamber holds three rugged tissue chamber modules (12 total), clear Teflon bags holding 30 ml of growth media, all positioned by a metal frame. Every 7 to 21 days (depending on growth rates), an astronaut uses a shrouded syringe and the bags' needleless injection ports to transfer a few cells to a fresh media bag, and to introduce a fixative so that the cells may be studied after flight. The design also lets the crew sample the media to measure glucose, gas, and pH levels, and to inspect cells with a microscope. The controller is monitored by the flight crew through a 23-cm (9-inch) color computer display on the face of the BSTC. This view shows the BTSC with the front panel open. 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.

  4. Mathematical modeling of ultrasound in tissue engineering: From bioreactors to the cellular scale

    Science.gov (United States)

    Louw, Tobias M.

    Tissue engineering seeks to provide a means to treat injuries that are beyond the body's natural ability to repair without the issues associated with allografts. Autologous cells are cultured in a bioreactor which controls the cellular environment (including mechanical stimulation) for optimal tissue growth. We investigate ultrasound as an effective means of mechanical stimulation by predicting the ultrasonic field in a bioreactor, as well as ultrasonic bioeffects at the cellular level. The Transfer Matrix Angular Spectrum Approach was found to be the most accurate and computationally efficient bioreactor model. Three critical factors influence experimental results: (1) the diameter of the tissue engineering scaffold greatly affects the ultrasonic field; (2) the position of the ultrasonic transducer and liquid level in the tissue culture well determines the maximum pressure amplitude in the bioreactor, but the pressure can be controlled by measuring the transducer input electrical impedance and manipulating the applied voltage; and (3) the position of pressure nodes are influenced by ultrasonic frequency and liquid level; this will affect the response of cells to applied ultrasound. On the cellular level, it was shown that chondrocytes respond to ultrasound with frequency dependence. A predicted resonance frequency near 5MHz matched experimental results showing maximum expression of load inducible genes at 5MHz. Mechanical stresses are concentrated near the nucleus at resonance, alluding to the possibility that the nucleus may directly sense ultrasonic stimulation. We postulate that ultrasound influences the transport of p-ERK to the nucleus or causes minor chromatin reorganization, leading to the observed frequency dependent gene expression. We linked in vitro ultrasonic stimulation to in vivo mechanical stimulation generated by natural movement. The chondrocyte's response to impact is under-damped, and the cell oscillates with a frequency close to the model

  5. Development and Characterization of a Parallelizable Perfusion Bioreactor for 3D Cell Culture

    Science.gov (United States)

    Egger, Dominik; Fischer, Monica; Clementi, Andreas; Ribitsch, Volker; Hansmann, Jan; Kasper, Cornelia

    2017-01-01

    The three dimensional (3D) cultivation of stem cells in dynamic bioreactor systems is essential in the context of regenerative medicine. Still, there is a lack of bioreactor systems that allow the cultivation of multiple independent samples under different conditions while ensuring comprehensive control over the mechanical environment. Therefore, we developed a miniaturized, parallelizable perfusion bioreactor system with two different bioreactor chambers. Pressure sensors were also implemented to determine the permeability of biomaterials which allows us to approximate the shear stress conditions. To characterize the flow velocity and shear stress profile of a porous scaffold in both bioreactor chambers, a computational fluid dynamics analysis was performed. Furthermore, the mixing behavior was characterized by acquisition of the residence time distributions. Finally, the effects of the different flow and shear stress profiles of the bioreactor chambers on osteogenic differentiation of human mesenchymal stem cells were evaluated in a proof of concept study. In conclusion, the data from computational fluid dynamics and shear stress calculations were found to be predictable for relative comparison of the bioreactor geometries, but not for final determination of the optimal flow rate. However, we suggest that the system is beneficial for parallel dynamic cultivation of multiple samples for 3D cell culture processes.

  6. Microliter-bioreactor array with buoyancy-driven stirring for human hematopoietic stem cell culture.

    Science.gov (United States)

    Luni, Camilla; Feldman, Hope C; Pozzobon, Michela; De Coppi, Paolo; Meinhart, Carl D; Elvassore, Nicola

    2010-08-11

    This work presents the development of an array of bioreactors where finely controlled stirring is provided at the microliter scale (100-300 mul). The microliter-bioreactor array is useful for performing protocol optimization in up to 96 parallel experiments of hematopoietic stem cell (HSC) cultures. Exploring a wide range of experimental conditions at the microliter scale minimizes cost and labor. Once the cell culture protocol is optimized, it can be applied to large-scale bioreactors for stem cell production at the clinical level. The controlled stirring inside the wells of a standard 96-well plate is provided by buoyancy-driven thermoconvection. The temperature and velocity fields within the culture volume are determined with numerical simulations. The numerical results are verified with experimental velocity measurements using microparticle image velocimetry (muPIV) and are used to define feasible experimental conditions for stem cell cultures. To test the bioreactor array's functionality, human umbilical cord blood-derived CD34(+) cells were cultured for 7 days at five different stirring conditions (0.24-0.58 mums) in six repeated experiments. Cells were characterized in terms of proliferation, and flow cytometry measurements of viability and CD34 expression. The microliter-bioreactor array demonstrates its ability to support HSC cultures under stirred conditions without adversely affecting the cell behavior. Because of the highly controlled operative conditions, it can be used to explore culture conditions where the mass transport of endogenous and exogenous growth factors is selectively enhanced, and cell suspension provided. While the bioreactor array was developed for culturing HSCs, its application can be extended to other cell types.

  7. NASA Bioreactor Demonstration System

    Science.gov (United States)

    2002-01-01

    Leland W. K. Chung (left), Director, Molecular Urology Therapeutics Program at the Winship Cancer Institute at Emory University, is principal investigator for the NASA bioreactor demonstration system (BDS-05). With him is Dr. Jun Shu, an assistant professor of Orthopedics Surgery from Kuming Medical University China. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. 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 Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. 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. Credit: Emory University.

  8. Visualizing medium and biodistribution in complex cell culture bioreactors using in vivo imaging.

    Science.gov (United States)

    Ratcliffe, E; Thomas, R J; Stacey, A J

    2014-01-01

    There is a dearth of technology and methods to aid process characterization, control and scale-up of complex culture platforms that provide niche micro-environments for some stem cell-based products. We have demonstrated a novel use of 3d in vivo imaging systems to visualize medium flow and cell distribution within a complex culture platform (hollow fiber bioreactor) to aid characterization of potential spatial heterogeneity and identify potential routes of bioreactor failure or sources of variability. This can then aid process characterization and control of such systems with a view to scale-up. Two potential sources of variation were observed with multiple bioreactors repeatedly imaged using two different imaging systems: shortcutting of medium between adjacent inlet and outlet ports with the potential to create medium gradients within the bioreactor, and localization of bioluminescent murine 4T1-luc2 cells upon inoculation with the potential to create variable seeding densities at different points within the cell growth chamber. The ability of the imaging technique to identify these key operational bioreactor characteristics demonstrates an emerging technique in troubleshooting and engineering optimization of bioreactor performance. © 2013 American Institute of Chemical Engineers.

  9. Impact of stirred suspension bioreactor culture on the differentiation of murine embryonic stem cells into cardiomyocytes

    Science.gov (United States)

    2011-01-01

    Background Embryonic stem cells (ESCs) can proliferate endlessly and are able to differentiate into all cell lineages that make up the adult organism. Under particular in vitro culture conditions, ESCs can be expanded and induced to differentiate into cardiomyocytes in stirred suspension bioreactors (SSBs). However, in using these systems we must be cognizant of the mechanical forces acting upon the cells. The effect of mechanical forces and shear stress on ESC pluripotency and differentiation has yet to be clarified. The purpose of this study was to investigate the impact of the suspension culture environment on ESC pluripotency during cardiomyocyte differentiation. Results Murine D3-MHC-neor ESCs formed embyroid bodies (EBs) and differentiated into cardiomyocytes over 25 days in static culture and suspension bioreactors. G418 (Geneticin) was used in both systems from day 10 to enrich for cardiomyocytes by eliminating non-resistant, undifferentiated cells. Treatment of EBs with 1 mM ascorbic acid and 0.5% dimethyl sulfoxide from day 3 markedly increased the number of beating EBs, which displayed spontaneous and cadenced contractile beating on day 11 in the bioreactor. Our results showed that the bioreactor differentiated cells displayed the characteristics of fully functional cardiomyocytes. Remarkably, however, our results demonstrated that the bioreactor differentiated ESCs retained their ability to express pluripotency markers, to form ESC-like colonies, and to generate teratomas upon transplantation, whereas the cells differentiated in adherent culture lost these characteristics. Conclusions This study demonstrates that although cardiomyocyte differentiation can be achieved in stirred suspension bioreactors, the addition of medium enhancers is not adequate to force complete differentiation as fluid shear forces appear to maintain a subpopulation of cells in a transient pluripotent state. The development of successful ESC differentiation protocols within

  10. Impact of stirred suspension bioreactor culture on the differentiation of murine embryonic stem cells into cardiomyocytes

    Directory of Open Access Journals (Sweden)

    Shafa Mehdi

    2011-12-01

    Full Text Available Abstract Background Embryonic stem cells (ESCs can proliferate endlessly and are able to differentiate into all cell lineages that make up the adult organism. Under particular in vitro culture conditions, ESCs can be expanded and induced to differentiate into cardiomyocytes in stirred suspension bioreactors (SSBs. However, in using these systems we must be cognizant of the mechanical forces acting upon the cells. The effect of mechanical forces and shear stress on ESC pluripotency and differentiation has yet to be clarified. The purpose of this study was to investigate the impact of the suspension culture environment on ESC pluripotency during cardiomyocyte differentiation. Results Murine D3-MHC-neor ESCs formed embyroid bodies (EBs and differentiated into cardiomyocytes over 25 days in static culture and suspension bioreactors. G418 (Geneticin was used in both systems from day 10 to enrich for cardiomyocytes by eliminating non-resistant, undifferentiated cells. Treatment of EBs with 1 mM ascorbic acid and 0.5% dimethyl sulfoxide from day 3 markedly increased the number of beating EBs, which displayed spontaneous and cadenced contractile beating on day 11 in the bioreactor. Our results showed that the bioreactor differentiated cells displayed the characteristics of fully functional cardiomyocytes. Remarkably, however, our results demonstrated that the bioreactor differentiated ESCs retained their ability to express pluripotency markers, to form ESC-like colonies, and to generate teratomas upon transplantation, whereas the cells differentiated in adherent culture lost these characteristics. Conclusions This study demonstrates that although cardiomyocyte differentiation can be achieved in stirred suspension bioreactors, the addition of medium enhancers is not adequate to force complete differentiation as fluid shear forces appear to maintain a subpopulation of cells in a transient pluripotent state. The development of successful ESC

  11. Novel disposable flexible bioreactor for Escherichia coli culture in orbital shaking incubator.

    Science.gov (United States)

    Yang, Ting; Huang, Yue; Han, Zhiqiang; Liu, Huitao; Zhang, Rui; Xu, Yuming

    2013-10-01

    Erlenmeyer flask or conical flask, usually made of glass, is widely used for laboratory scale suspension culture of microorganism, such as Escherichia Coli and yeast. Due to being non-disposable culture vessel, it has to be cleaned, packaged and sterilized prior to use, which are time, labor and energy consuming work, and has the potential risk of cross-contamination. Despite the rigid plastic conical flasks are possible for single use, they are not economically effective and produce more waste. To overcome these drawbacks, here we successfully developed a novel disposable flexible bioreactor with a plastic film through a thermo-fusion technique. With a triangular pyramid shape, the bioreactor enabled itself to keep a three-dimensional internal space without needing air inflation and well adapted to the commercial available orbital shaker. Unlike the conventional rigid conical flasks and other reported flexible flasks, which had to be fixed in the shaker, the flexible bioreactor could keep sitting on the silicone pad-carpeted platform of the orbital shaker steadily without any fixation needed at the shaking speeds below 250 rpm, thus making it simple to handle. Compared with the traditional conical glass flasks, the innovative flexible bioreactors achieved a significant higher efficiency in bacteria growth and oxygen transfer rates. In conclusion, the novel flexible bioreactor is an ideal disposable culture vessel for microorganism suspension culture at laboratory scale and holds a promising potential to replace the glass flask and rigid plastic flask in the future.

  12. Suspension cell culture in microgravity and development of a space bioreactor

    Science.gov (United States)

    Morrison, Dennis R.

    1987-01-01

    NASA has methodically developed unique suspension type cell and recovery apparatus culture systems for bioprocess technology experiments and production of biological products in microgravity. The first space bioreactor has been designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small (500 ml) bioreactor is being constructed for flight experiments in the Shuttle middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption, and control of low shear stress on cells.

  13. On modelling, monitoring and control of fixed bed bioreactors

    Energy Technology Data Exchange (ETDEWEB)

    Tali-Maamar, N.; Babary, J.P. [Centre National de la Recherche Scientifique (CNRS), 31 - Toulouse (France); Dochain, D. [Universite Catholique de Louvain (UCL), Louvain-la-Neuve (Belgium)

    1995-12-31

    The working of a denitrification fixed bed bioreactor strongly depends on time and location of observing point, resulting in partial differential equations systems for the functional model. This project tries to extend the application of adaptative linearizing control schemes to non linear distributed parameters bio process. A second goal is to optimize the sensors position. (D.L.) 23 refs.

  14. Methyl jasmonate induced overproduction of eleutherosides in somatic embryos of Eleutherococcus senticosus cultured in bioreactors

    OpenAIRE

    Shohael,Abdullah Mohammad; Murthy,Hosakatte Niranjana; Hahn, Eun-Joo; Paek,Kee-Yoeup

    2007-01-01

    This study was concentrated on the production of eleutherosides and chlorogenic acid in embryogenic suspension cultures of Eleutherococcus senticosus by exposing them to different concentrations (50-400 µM) of methyl jasmonate (MJ) during the culture period. In the bioreactor cultures, eleutheroside content increased significantly by elicitation of MJ, however, the fresh weight, dry weight and growth ratio of embryos was strongly inhibited by increasing MJ concentrations. The highest total el...

  15. Mathematical modelling of cell layer growth in a hollow fibre bioreactor.

    Science.gov (United States)

    Chapman, Lloyd A C; Whiteley, Jonathan P; Byrne, Helen M; Waters, Sarah L; Shipley, Rebecca J

    2017-04-07

    Generating autologous tissue grafts of a clinically useful volume requires efficient and controlled expansion of cell populations harvested from patients. Hollow fibre bioreactors show promise as cell expansion devices, owing to their potential for scale-up. However, further research is required to establish how to specify appropriate hollow fibre bioreactor operating conditions for expanding different cell types. In this study we develop a simple model for the growth of a cell layer seeded on the outer surface of a single fibre in a perfused hollow fibre bioreactor. Nutrient-rich culture medium is pumped through the fibre lumen and leaves the bioreactor via the lumen outlet or passes through the porous fibre walls and cell layer, and out via ports on the outer wall of the extra-capillary space. Stokes and Darcy equations for fluid flow in the fibre lumen, fibre wall, cell layer and extra-capillary space are coupled to reaction-advection-diffusion equations for oxygen and lactate transport through the bioreactor, and to a simple growth law for the evolution of the free boundary of the cell layer. Cells at the free boundary are assumed to proliferate at a rate that increases with the local oxygen concentration, and to die and detach from the layer if the local fluid shear stress or lactate concentration exceed critical thresholds. We use the model to predict operating conditions that maximise the cell layer growth for different cell types. In particular, we predict the optimal flow rate of culture medium into the fibre lumen and fluid pressure imposed at the lumen outlet for cell types with different oxygen demands and fluid shear stress tolerances, and compare the growth of the cell layer when the exit ports on the outside of the bioreactor are open with that when they are closed. Model simulations reveal that increasing the inlet flow rate and outlet fluid pressure increases oxygen delivery to the cell layer and, therefore, the growth rate of cells that are

  16. Bioreactor rotating wall vessel

    Science.gov (United States)

    2001-01-01

    The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. 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. Cell constructs grown in a rotating bioreactor on Earth (left) eventually become too large to stay suspended in the nutrient media. In the microgravity of orbit, the cells stay suspended. Rotation then is needed for gentle stirring to replenish the media around the cells.

  17. Bioreactor rotating wall vessel

    Science.gov (United States)

    2001-01-01

    The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. 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. Cell constructs grown in a rotating bioreactor on Earth (left) eventually become too large to stay suspended in the nutrient media. In the microgravity of orbit, the cells stay suspended. Rotation then is needed for gentle stirring to replenish the media around the cells.

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

    Directory of Open Access Journals (Sweden)

    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.

  19. 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...

  20. Bioreactor production of secondary metabolites from cell cultures of periwinkle and sandalwood.

    Science.gov (United States)

    Valluri, Jagan V

    2009-01-01

    A bench-top bioreactor allowing continuous extraction of secondary metabolites is designed for Catharanthus roseus L. (G.) Don (periwinkle) and Santalum album L. (sandalwood) plant cell suspensions. Periwinkle cell cultures are exposed to biotic elicitors (Aspergillus niger, crude chitin) and abiotic elicitors (mannitol, methyl jasmonate) to induce alkaloid production. Whereas most of the biotic elicitors are effective when added on day 15 of culture, the abiotic elicitors are effective when added on day 20. The use of trans-cinnamic acid, an inhibitor of phenylalanine ammonia lyase (PAL) activity, results in significant increase in the alkaloid production of periwinkle cell cultures. Exposure of the cells to mannitol-induced osmotic stress produced marked increment in the total alkaloid production. When biotic and abiotic stress treatments are applied sequentially, an additive effect in alkaloid accumulation is observed. Although no essential oils are detected, secondary metabolites in the form of phenolics are produced by the sandalwood cell cultures in the bioreactor environment. The use of morphologic modification such as organ cultures and transformed cultures is believed to be required for both production and storage of essential oil constituents in sandalwood. The present chapter demonstrates that periwinkle and sandalwood cell suspensions could be developed and successfully cultured in a modified air-lift bioreactor. The exploitation of variant cell strains and biotransformation of added precursors can certainly improve the use of periwinkle and sandalwood cell cultures for the bioproduction of desired compounds.

  1. 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...

  2. 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

  3. Production of human alpha-1-antitrypsin from transgenic rice cell culture in a membrane bioreactor.

    Science.gov (United States)

    McDonald, Karen A; Hong, Lo Ming; Trombly, David M; Xie, Qing; Jackman, Alan P

    2005-01-01

    Transgenic plant cell cultures offer a number of advantages over alternative host expression systems, but so far relatively low product concentrations have been achieved. In this study, transgenic rice cells are used in a two-compartment membrane bioreactor (CELLine 350, Integra Biosciences) for the production of recombinant alpha-1-antitrypsin (rAAT). Expression of rAAT is controlled by the rice alpha-amylase (RAmy3D) promoter, which is induced in the absence of sugar. The extracellular product is retained in the bioreactor's relatively small cell compartment, thereby increasing product concentration. Due to the packed nature of the cell aggregates in the cell compartment, a clarified product solution can be withdrawn from the bioreactor. Active rAAT reached levels of 100-247 mg/L (4-10% of the total extracellular protein) in the cell compartment at 5-6 days postinduction, and multiple inductions of the RAmy3D promoter were demonstrated.

  4. 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

  5. Long term organ culture of human prostate tissue in a NASA-designed rotating wall bioreactor

    Science.gov (United States)

    Margolis, L.; Hatfill, S.; Chuaqui, R.; Vocke, C.; Emmert-Buck, M.; Linehan, W. M.; Duray, P. H.

    1999-01-01

    PURPOSE: To maintain ex vivo integral prostatic tissue including intact stromal and ductal elements using the NASA-designed Rotating Wall Vessel (RWV) which maintains colocalized cells in an environment that promotes both three-dimensional cellular interactions together with the uniform mass transfer of nutrients and metabolic wastes. MATERIALS AND METHODS: Samples of normal prostate were obtained as a byproduct of transurethral prostatectomy or needle biopsy. Prostatic tissue dissected into small 1 x 1 mm. blocks was cultured in the Rotating Wall Vessel (RWV) Bioreactor for various time periods and analyzed using histological, immunochemical, and total cell RNA assays. RESULTS: We report the long term maintenance of benign explanted human prostate tissue grown in simple culture medium, under the simulated microgravity conditions afforded by the RWV bioreactor. Mesenchymal stromal elements including blood vessels and architecturally preserved tubuloglandular acini were maintained for a minimum of 28 days. Cytokeratins, vimentin and TGF-beta2 receptor and ligand were preserved through the entire culture period as revealed by immunocytochemistry. Prostatic acid phosphatase (PAP) was continuously expressed during the culture period, although somewhat decreased. Prostatic specific antigen (PSA) and its transcript were down regulated over time of culture. Prostatic carcinoma cells from the TSU cell line were able to invade RWV-cultured benign prostate tissue explants. CONCLUSIONS: The RWV bioreactor represents an additional new technology for culturing prostate tissue for further investigations concerning the basic physiology and pathobiology of this clinically important tissue.

  6. Large-Scale Plantlet Conversion and Ex Vitro Transplantation Efficiency of Siberian Ginseng by Bioreactor Culture

    Directory of Open Access Journals (Sweden)

    Jingli Yang

    2013-01-01

    Full Text Available To achieve large-scale low-cost ex vitro acclimatization of Siberian ginseng plants, heart- and torpedo-shaped secondary somatic embryos (SEs induced from germinated SEs on agar medium were collected and then inoculated to 10-l bubble column bioreactor, respectively. For plantlet conversion, inoculation of torpedo-shaped secondary SEs was more effective than heart-shaped SEs. TS2 (culture of torpedo-shaped SEs in a bioreactor with a 2-week subculture interval plantlets had a higher root number and leaf number and larger leaf area than did HS3 (culture of heart-shaped SEs in a bioreactor with a 3-week subculture interval and HS2 (culture of heart-shaped SEs in a bioreactor with a 2-week subculture interval plantlets. Of these converted plants, TS2 plantlets had higher survival rate (83.7% and growth characteristics after transplantation in a simple shed covered with a 50% sunshade net only for 6 months. TS2 plantlets also showed significantly lower H2O2 content and significantly increased superoxide dismutase (SOD, glutathione peroxidase (GPX, and glutathione transferase (GST expression levels as compared to HS2 plants when exposure to ex vitro conditions.

  7. Novel micro-bioreactor high throughput technology for cell culture process development: Reproducibility and scalability assessment of fed-batch CHO cultures.

    Science.gov (United States)

    Amanullah, Ashraf; Otero, Jose Manuel; Mikola, Mark; Hsu, Amy; Zhang, Jinyou; Aunins, John; Schreyer, H Brett; Hope, James A; Russo, A Peter

    2010-05-01

    With increasing timeline pressures to get therapeutic and vaccine candidates into the clinic, resource intensive approaches such as the use of shake flasks and bench-top bioreactors may limit the design space for experimentation to yield highly productive processes. The need to conduct large numbers of experiments has resulted in the use of miniaturized high-throughput (HT) technology for process development. One such high-throughput system is the SimCell platform, a robotically driven, cell culture bioreactor system developed by BioProcessors Corp. This study describes the use of the SimCell micro-bioreactor technology for fed-batch cultivation of a GS-CHO transfectant expressing a model IgG4 monoclonal antibody. Cultivations were conducted in gas-permeable chambers based on a micro-fluidic design, with six micro-bioreactors (MBs) per micro-bioreactor array (MBA). Online, non-invasive measurement of total cell density, pH and dissolved oxygen (DO) was performed. One hundred fourteen parallel MBs (19 MBAs) were employed to examine process reproducibility and scalability at shake flask, 3- and 100-L bioreactor scales. The results of the study demonstrate that the SimCell platform operated under fed-batch conditions could support viable cell concentrations up to least 12 x 10(6) cells/mL. In addition, both intra-MB (MB to MB) as well as intra-MBA (MBA to MBA) culture performance was found to be highly reproducible. The intra-MB and -MBA variability was calculated for each measurement as the coefficient of variation defined as CV (%) = (standard deviation/mean) x 100. The % CV values for most intra-MB and intra-MBA measurements were generally under 10% and the intra-MBA values were slightly lower than those for intra-MB. Cell growth, process parameters, metabolic and protein titer profiles were also compared to those from shake flask, bench-top, and pilot scale bioreactor cultivations and found to be within +/-20% of the historical averages.

  8. Estimation of dissolved carbon dioxide stripping in a large bioreactor using model medium.

    Science.gov (United States)

    Matsunaga, Naoki; Kano, Kenjiro; Maki, Yasuyuki; Dobashi, Toshiaki

    2009-04-01

    Dissolved carbon dioxide (dCO(2)) accumulation is one of the most serious problems in the scale-up of industrial cell culture. To predict the effects of dCO(2) stripping in different culture conditions and at different scales, we examined a method of estimation of dCO(2) stripping using a model medium. The operational parameters (e.g., sparging and agitation rate) and the size of the bioreactor (working volume: 80 L, 500 L, 2000 L; aspect ratio: 1.0 approximately 1.6) were varied, and the model medium was prepared by adjusting pH, density, viscosity, surface tension, and buffer conditions. dCO(2) stripping efficiency was evaluated using the index k(L)a(CO)((2)), which was defined in accordance with the volumetric oxygen transfer coefficient k(L)a. The model medium exhibited dCO(2) stripping behavior similar to real culture medium in all experimental conditions tested. It is expected that the use of the model medium to estimate dCO(2) stripping in real cultures will be valuable for determining the culture conditions in bioreactors in scale-up.

  9. Bioreactor for application of subatmospheric pressure to three-dimensional cell culture.

    Science.gov (United States)

    Wilkes, Robert P; McNulty, Amy K; Feeley, Teri D; Schmidt, Marisa A; Kieswetter, Kris

    2007-12-01

    Vacuum-assisted closure (VAC) negative pressure wound therapy (NPWT) is a highly successful and widely used treatment modality for wound healing, although no apparatus exists to monitor the effects of subatmospheric pressure application in vitro. Such an apparatus is desirable to better understand the biological effects of this therapy and potentially improve upon them. This article describes the development and validation of a novel bioreactor that permits such study. Tissue analogues consisting of 3-dimensional fibroblast-containing fibrin clots were cultured in off-the-shelf disposable cell culture inserts and multi-well plates that were integrated into the bioreactor module. Negative pressure dressings, commercialized for wound therapy, were placed on top of the culture, and subatmospheric pressure was applied to the dressing. Cultures were perfused with media at controlled physiologic wound exudate flow rates. The design of this bioreactor permits observation of the culture using an inverted microscope in brightfield and fluorescence modes and sustained incubation of the system in a 5% carbon dioxide atmosphere. This closed-system mimics the wound micro-environment under VAC NPWT. Matrix compression occurs as the subatmospheric pressure draws the dressing material down. At the contact zone, surface undulations were clearly evident on the fibroblast-containing tissue analogues at 24 h and appeared to correspond to the dressing microstructure. The bioreactor design, consisting of sterilizable machined plastics and disposable labware, can be easily scaled to multiple units. Validation experiments show that cell survival in this system is comparable with that seen in cells grown in static tissue culture. After application of VAC NPWT, cell morphology changed, with cells appearing thicker and with an organized actin cytoskeleton. The development and validation of this new culture system establishes a stable platform for in vitro investigations of subatmospheric

  10. Bioreactor concepts for cell culture-based viral vaccine production.

    Science.gov (United States)

    Gallo-Ramírez, Lilí Esmeralda; Nikolay, Alexander; Genzel, Yvonne; Reichl, Udo

    2015-01-01

    Vaccine manufacturing processes are designed to meet present and upcoming challenges associated with a growing vaccine market and to include multi-use facilities offering a broad portfolio and faster reaction times in case of pandemics and emerging diseases. The final products, from whole viruses to recombinant viral proteins, are very diverse, making standard process strategies hardly universally applicable. Numerous factors such as cell substrate, virus strain or expression system, medium, cultivation system, cultivation method, and scale need consideration. Reviewing options for efficient and economical production of human vaccines, this paper discusses basic factors relevant for viral antigen production in mammalian cells, avian cells and insect cells. In addition, bioreactor concepts, including static systems, single-use systems, stirred tanks and packed-beds are addressed. On this basis, methods towards process intensification, in particular operational strategies, the use of perfusion systems for high product yields, and steps to establish continuous processes are introduced.

  11. Mathematical modelling of flow and transport processes in tissue engineering bioreactors

    Science.gov (United States)

    Waters, Sarah; Pearson, Natalie; Oliver, James; Shipley, Rebecca

    2014-11-01

    To artificially engineer tissues numerous biophysical and biochemical processes must be integrated to produce tissues with the desired in vivo properties. Tissue engineering bioreactors are cell culture systems which aim to mimic the in vivo environment. We consider a hollow fibre membrane bioreactor (HFMB), which utilises fluid flow to enhance the delivery of growth factors and nutrients to, and metabolite removal from, the cells, as well as provide appropriate mechanical stimuli to the cells. Biological tissues comprise a wide variety of interacting components, and multiphase models provide a natural framework to investigate such interactions. We present a suite of mathematical models (capturing different experimental setups) which consider the fluid flow, solute transport, and cell yield and distribution within a HFMB. The governing equations are simplified by exploiting the slender geometry of the bioreactor system, so that, e.g., lubrication theory may be used to describe flow in the lumen. We interrogate the models to illustrate typical behaviours of each setup in turn, and highlight the dependence of results on key experimentally controllable parameter values. Once validated, such models can be used to inform and direct future experiments.

  12. NASA Classroom Bioreactor

    Science.gov (United States)

    Scully, Robert

    2004-01-01

    Exploration of space provides a compelling need for cell-based research into the basic mechanisms that underlie the profound changes that occur in terrestrial life that is transitioned to low gravity environments. Toward that end, NASA developed a rotating bioreactor in which cells are cultured while continuously suspended in a cylinder in which the culture medium rotates with the cylinder. The randomization of the gravity vector accomplished by the continuous rotation, in a low shear environment, provides an analog of microgravity. Because cultures grown in bioreactors develop structures and functions that are much closer to those exhibited by native tissue than can be achieved with traditional culture methods, bioreactors have contributed substantially to advancing research in the fields of cancer, diabetes, infectious disease modeling for vaccine production, drug efficacy, and tissue engineering. NASA has developed a Classroom Bioreactor (CB) that is built from parts that are easily obtained and assembled, user-friendly and versatile. It can be easily used in simple school settings to examine the effect cultures of seeds or cells. An educational brief provides assembly instructions and lesson plans that describes activities in science, math and technology that explore free fall, microgravity, orbits, bioreactors, structure-function relationships and the scientific method.

  13. NASA Classroom Bioreactor

    Science.gov (United States)

    Scully, Robert

    2004-01-01

    Exploration of space provides a compelling need for cell-based research into the basic mechanisms that underlie the profound changes that occur in terrestrial life that is transitioned to low gravity environments. Toward that end, NASA developed a rotating bioreactor in which cells are cultured while continuously suspended in a cylinder in which the culture medium rotates with the cylinder. The randomization of the gravity vector accomplished by the continuous rotation, in a low shear environment, provides an analog of microgravity. Because cultures grown in bioreactors develop structures and functions that are much closer to those exhibited by native tissue than can be achieved with traditional culture methods, bioreactors have contributed substantially to advancing research in the fields of cancer, diabetes, infectious disease modeling for vaccine production, drug efficacy, and tissue engineering. NASA has developed a Classroom Bioreactor (CB) that is built from parts that are easily obtained and assembled, user-friendly and versatile. It can be easily used in simple school settings to examine the effect cultures of seeds or cells. An educational brief provides assembly instructions and lesson plans that describes activities in science, math and technology that explore free fall, microgravity, orbits, bioreactors, structure-function relationships and the scientific method.

  14. Bioreactor engineering using disposable technology for enhanced production of hCTLA4Ig in transgenic rice cell cultures.

    Science.gov (United States)

    Kwon, Jun-Young; Yang, Yong-Suk; Cheon, Su-Hwan; Nam, Hyung-Jin; Jin, Gi-Hong; Kim, Dong-Il

    2013-09-01

    Two kinds of disposable bioreactors, air-lift disposable bioreactors (ADB) and wave disposable bioreactors (WDB) were compared with stirred-tank reactors (5-L STR). These bioreactors were successfully applied to transgenic rice cell cultures for the production of recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig). In both systems, a fed-batch culture method was used to produce hCTLA4Ig efficiently by feeding concentrated amino acids and production levels were enhanced when dissolved oxygen (DO) level was regulated at 30% using pure oxygen sparging. Agitation and aeration rate during cultivation in ADB and WDB were determined by the same mixing time. The results in both disposable bioreactors showed similar values in maximum cell density (11.9 gDCW/L and 12.6 gDCW/L), doubling time (4.8- and 5.0-day), and maximum hCTLA4Ig concentration (43.7 and 43.3 mg/L). Relatively higher cell viability was sustained in the ADB whereas hCTLA4Ig productivity was 1.2-fold higher than that in WDB. The productivity was improved by increasing aeration rate (0.2 vvm). Overall, our experiments demonstrate pneumatically driven disposable bioreactors are applicable for the production of recombinant proteins in plant cell cultures. These results will be useful for development and scale-up studies of disposable bioreactor systems for transgenic plant cell cultures.

  15. Multicompartmental Hollow-Fiber-Based Bioreactors for Dynamic Three-Dimensional Perfusion Culture.

    Science.gov (United States)

    Schmelzer, Eva; Gerlach, Jörg C

    2016-01-01

    The creation of larger-scale three-dimensional tissue constructs depends on proper medium mass and gas exchange, as well as removal of metabolites, which cannot be achieved in conventional static two-dimensional petri dish culture. In cultures of tissue-density this problem can be addressed by decentral perfusion through artificial micro-capillaries. While the static medium exchange in petri dishes leads to metabolite peaks, perfusion culture provides a dynamic medium supply, thereby preventing non-physiological peaks. To overcome the limitations of conventional static two-dimensional culture, a three-dimensional perfusion bioreactor technology has been developed, providing decentral and high-performance mass exchange as well as integral oxygenation. Similar to organ systems in vivo, the perfusion with medium provides nutrition and removes waste metabolites, and the perfusion with gas delivers oxygen and carbon dioxide for pH regulation. Such bioreactors are available at various dimensions ranging from 0.2 to 800 mL cell compartment volumes (manufactured by StemCell Systems, Berlin, Germany). Here, we describe in detail the setup and maintenance of a small-scale 4-chamber bioreactor with its tubing circuit and perfusion system.

  16. Culture of human mesenchymal stem cells on microcarriers in a 5 l stirred-tank bioreactor.

    Science.gov (United States)

    Rafiq, Qasim A; Brosnan, Kathryn M; Coopman, Karen; Nienow, Alvin W; Hewitt, Christopher J

    2013-08-01

    For the first time, fully functional human mesenchymal stem cells (hMSCs) have been cultured at the litre-scale on microcarriers in a stirred-tank 5 l bioreactor, (2.5 l working volume) and were harvested via a potentially scalable detachment protocol that allowed for the successful detachment of hMSCs from the cell-microcarrier suspension. Over 12 days, the dissolved O2 concentration was >45 % of saturation and the pH between 7.2 and 6.7 giving a maximum cell density in the 5 l bioreactor of 1.7 × 10(5) cells/ml; this represents >sixfold expansion of the hMSCs, equivalent to that achievable from 65 fully-confluent T-175 flasks. During this time, the average specific O2 uptake of the cells in the 5 l bioreactor was 8.1 fmol/cell h and, in all cases, the 5 l bioreactors outperformed the equivalent 100 ml spinner-flasks run in parallel with respect to cell yields and growth rates. In addition, yield coefficients, specific growth rates and doubling times were calculated for all systems. Neither the upstream nor downstream bioprocessing unit operations had a discernible effect on cell quality with the harvested cells retaining their immunophenotypic markers, key morphological features and differentiation capacity.

  17. Critical Review of Membrane Bioreactor Models

    DEFF Research Database (Denmark)

    Naessens, W.; Maere, T.; Ratkovich, Nicolas Rios;

    2012-01-01

    modelling. In this paper, the vast literature on hydrodynamic and integrated modelling in MBR is critically reviewed. Hydrodynamic models are used at different scales and focus mainly on fouling and only little on system design/optimisation. Integrated models also focus on fouling although the ones...

  18. Application of multivariate analysis and mass transfer principles for refinement of a 3-L bioreactor scale-down model--when shake flasks mimic 15,000-L bioreactors better.

    Science.gov (United States)

    Ahuja, Sanjeev; Jain, Shilpa; Ram, Kripa

    2015-01-01

    Characterization of manufacturing processes is key to understanding the effects of process parameters on process performance and product quality. These studies are generally conducted using small-scale model systems. Because of the importance of the results derived from these studies, the small-scale model should be predictive of large scale. Typically, small-scale bioreactors, which are considered superior to shake flasks in simulating large-scale bioreactors, are used as the scale-down models for characterizing mammalian cell culture processes. In this article, we describe a case study where a cell culture unit operation in bioreactors using one-sided pH control and their satellites (small-scale runs conducted using the same post-inoculation cultures and nutrient feeds) in 3-L bioreactors and shake flasks indicated that shake flasks mimicked the large-scale performance better than 3-L bioreactors. We detail here how multivariate analysis was used to make the pertinent assessment and to generate the hypothesis for refining the existing 3-L scale-down model. Relevant statistical techniques such as principal component analysis, partial least square, orthogonal partial least square, and discriminant analysis were used to identify the outliers and to determine the discriminatory variables responsible for performance differences at different scales. The resulting analysis, in combination with mass transfer principles, led to the hypothesis that observed similarities between 15,000-L and shake flask runs, and differences between 15,000-L and 3-L runs, were due to pCO2 and pH values. This hypothesis was confirmed by changing the aeration strategy at 3-L scale. By reducing the initial sparge rate in 3-L bioreactor, process performance and product quality data moved closer to that of large scale. © 2015 American Institute of Chemical Engineers.

  19. THE USE OF CONTINUOUS, TEMPORARY IMMERSION BIOREACTOR SYSTEM AND SEMISOLID CULTURE MEDIUM FOR THE PRODUCTION OF Eucalyptus camaldulensis CLONES

    Directory of Open Access Journals (Sweden)

    Evânia Galvão Mendonça

    2016-01-01

    Full Text Available The plant micro-propagation in bioreactor systems is regarded as one way to reduce cost by automation and production scheduling. This research was carried out in order to obtain an efficient procedure for clone production of Eucalyptus camaldulensis on different types of bioreactor including continuous and temporary immersion bioreactor. To do so, the apical meristems (1 mm and the apical meristems with adjacent tissue (2,5 mm were used as initial explants. These tissues were cultured, for 60 days, in semisolid culture medium supplemented with 1 mg L -1 indole acetic acid (IAA and 0.32 mg L -1 benzylaminopurine (BA. After 60 days, the meristems with adjacent tissue were transferred to a continuous immersion bioreactor and maintained in dark or light conditions. In order to verify the effect of the explant source on bioreactor multiplication, the explants subcultured from meristems multiplied in semisolid culture medium and the meristems multiplied in continuous immersion bioreactor were tested and maintained in dark conditions. After establishing this parameters, the multiplication experiments were carried out in continuous and temporary immersion and the multiplied explants were then rooted in MS medium supplemented with 0, 2, 4, 8 and 20 mg L -1 indole butyric acid (IBA and kept in the dark or under controlled lighting conditions. After that, the rooting the plants were acclimatized in mist chamber. The meristem with adjacent tissue favored a greater number of buds/explants. The continuous immersion bioreactor in the dark provided higher shoots number and multiplication rate. The rooting was better on culture medium without auxin and kept in the dark for 15 days or the culture medium supplemented with auxin and maintained under light with 100% plantlet rooting. The Eucalyptus camaldulensis acclimatization was efficient, with high survival rate (76%. It was possible to establish the procedure for bioreactor micro-propagation of Eucalyptus

  20. A Rotating Bioreactor for Scalable Culture and Differentiation of Respiratory Epithelium.

    Science.gov (United States)

    Raredon, Micha Sam Brickman; Ghaedi, Mahboobe; Calle, Elizabeth A; Niklason, Laura E

    2015-10-01

    Respiratory epithelium is difficult to grow in vitro, as it requires a well-maintained polarizing air-liquid interface (ALI) to maintain differentiation. Traditional methods rely on permeable membrane culture inserts, which are difficult to work with and are ill-suited for the production of large numbers of cells, such as the quantities required for cell-based clinical therapies. Herein, we investigate an alternative form of culture in which the cells are placed on a porous substrate that is continuously rolled, such that the monolayer of cells is alternately submerged in media or apically exposed to air. Our prototype bioreactor is reliable for up to 21 days of continuous culture and is designed for scale-up for large-scale cell culture with continuous medium and gas exchange. Normal human bronchial epithelial (NHBE) cells were cultured on an absorbent substrate in the reactor for periods of 7, 14, and 21 days and were compared to static controls that were submerged in media. Quantification by immunohistochemistry and quantitative PCR of markers specific to differentiated respiratory epithelium indicated increased cilia, mucous production, and tight junction formation in the rolled cultures, compared to static. Together with scanning electron microscopy and paraffin histology, the data indicate that the intermittent ALI provided by the rolling bioreactor promotes a polarized epithelial phenotype over a period of 21 days.

  1. Hybrid modeling of xanthan gum bioproduction in batch bioreactor.

    Science.gov (United States)

    Zabot, Giovani L; Mecca, Jaqueline; Mesomo, Michele; Silva, Marceli F; Prá, Valéria Dal; de Oliveira, Débora; Oliveira, J Vladimir; Castilhos, Fernanda; Treichel, Helen; Mazutti, Marcio A

    2011-10-01

    This work is focused on hybrid modeling of xanthan gum bioproduction process by Xanthomonas campestris pv. mangiferaeindicae. Experiments were carried out to evaluate the effects of stirred speed and superficial gas velocity on the kinetics of cell growth, lactose consumption and xanthan gum production in a batch bioreactor using cheese whey as substrate. A hybrid model was employed to simulate the bio-process making use of an artificial neural network (ANN) as a kinetic parameter estimator for the phenomenological model. The hybrid modeling of the process provided a satisfactory fitting quality of the experimental data, since this approach makes possible the incorporation of the effects of operational variables on model parameters. The applicability of the validated model was investigated, using the model as a process simulator to evaluate the effects of initial cell and lactose concentration in the xanthan gum production.

  2. Geosmin and Related Volatiles in Bioreactor-Cultured Streptomyces citreus CBS 109.60

    Science.gov (United States)

    Pollak, F. C.; Berger, R. G.

    1996-01-01

    Streptomyces citreus CBS 109.60 produced geosmin and a complex pattern of other volatile compounds during cultivation in a 2.5-liter laboratory bioreactor. Volatiles were isolated from disrupted cells, from the culture medium, and from the waste air of the bioreactor by adsorption on Lewatit OC 1064MD. Quantitative and qualitative analyses were carried out using capillary gas chromatography and coupled gas chromatography-mass spectroscopy. S. citreus produced 56 volatile compounds, which were mainly terpenoids but also included aliphatic ketones, alcohols, esters, pyrazines, furan(on)es, and aromatic types during the growth phase. The major components were geosmin and a germacradienol. A biosynthetic pathway for geosmin including eudesmanolides is proposed. PMID:16535293

  3. Engineering Tendon: Scaffolds, Bioreactors, and Models of Regeneration

    Directory of Open Access Journals (Sweden)

    Daniel W. Youngstrom

    2016-01-01

    Full Text Available Tendons bridge muscle and bone, translating forces to the skeleton and increasing the safety and efficiency of locomotion. When tendons fail or degenerate, there are no effective pharmacological interventions. The lack of available options to treat damaged tendons has created a need to better understand and improve the repair process, particularly when suitable autologous donor tissue is unavailable for transplantation. Cells within tendon dynamically react to loading conditions and undergo phenotypic changes in response to mechanobiological stimuli. Tenocytes respond to ultrastructural topography and mechanical deformation via a complex set of behaviors involving force-sensitive membrane receptor activity, changes in cytoskeletal contractility, and transcriptional regulation. Effective ex vivo model systems are needed to emulate the native environment of a tissue and to translate cell-matrix forces with high fidelity. While early bioreactor designs have greatly expanded our knowledge of mechanotransduction, traditional scaffolds do not fully model the topography, composition, and mechanical properties of native tendon. Decellularized tendon is an ideal scaffold for cultivating replacement tissue and modeling tendon regeneration. Decellularized tendon scaffolds (DTS possess high clinical relevance, faithfully translate forces to the cellular scale, and have bulk material properties that match natural tissue. This review summarizes progress in tendon tissue engineering, with a focus on DTS and bioreactor systems.

  4. DESIGNING SULFATE-REDUCING BACTERIA FIELD BIOREACTORS USING THE BEST MODEL

    Science.gov (United States)

    BEST (bioreactor economics, size and time of operation) is a spreadsheet-based model that is used in conjunction with a public domain computer software package, PHREEQCI. BEST is intended to be used in the design process of sulfate-reducing bacteria (SRB)field bioreactors to pas...

  5. Hemoglobin Regulates the Metabolic, Synthetic, Detoxification, and Biotransformation Functions of Hepatoma Cells Cultured in a Hollow Fiber Bioreactor

    Science.gov (United States)

    Chen, Guo

    2010-01-01

    Hepatic hollow fiber (HF) bioreactors constitute one type of extracorporeal bioartificial liver assist device (BLAD). Ideally, cultured hepatocytes in a BLAD should closely mimic the in vivo oxygenation environment of the liver sinusoid to yield a device with optimal performance. However, most BLADs, including hepatic HF bioreactors, suffer from O2 limited transport toward cultured hepatocytes, which reduces their performance. We hypothesize that supplementation of hemoglobin-based O2 carriers into the circulating cell culture medium of hepatic HF bioreactors is a feasible and effective strategy to improve bioreactor oxygenation and performance. We examined the effect of bovine hemoglobin (BvHb) supplementation (15 g/L) in the circulating cell culture medium of hepatic HF bioreactors on hepatocyte proliferation, metabolism, and varied liver functions, including biosynthesis, detoxification, and biotransformation. It was observed that BvHb supplementation supported the maintenance of a higher cell mass in the extracapillary space, improved hepatocyte metabolic efficiency (i.e., hepatocytes consumed much less glucose), improved hepatocyte capacity for drug metabolism, and conserved both albumin synthesis and ammonia detoxification functions compared to controls (no BvHb supplementation) under the same experimental conditions. PMID:20528678

  6. 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

    2017-09-05

    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 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.

  7. MALDI-TOF characterization of hGH1 produced by hairy root cultures of Brassica oleracea var. italica grown in an airlift with mesh bioreactor.

    Science.gov (United States)

    López, Edgar García; Ramírez, Emma Gloria Ramos; Gúzman, Octavio Gómez; Calva, Graciano Calva; Ariza-Castolo, Armando; Pérez-Vargas, Josefina; Rodríguez, Herminia Guadalupe Martínez

    2014-01-01

    Expression systems based on plant cells, tissue, and organ cultures have been investigated as an alternative for production of human therapeutic proteins in bioreactors. In this work, hairy root cultures of Brassica oleracea var. italica (broccoli) were established in an airlift with mesh bioreactor to produce isoform 1 of the human growth hormone (hGH1) as a model therapeutic protein. The hGH1 cDNA was cloned into the pCAMBIA1105.1 binary vector to induce hairy roots in hypocotyls of broccoli plantlets via Agrobacterium rhizogenes. Most of the infected plantlets (90%) developed hairy roots when inoculated before the appearance of true leaves, and keeping the emerging roots attached to hypocotyl explants during transfer to solid Schenk and Hildebrandt medium. The incorporation of the cDNA into the hairy root genome was confirmed by PCR amplification from genomic DNA. The expression and structure of the transgenic hGH1 was assessed by ELISA, western blot, and MALDITOF-MS analysis of the purified protein extracted from the biomass of hairy roots cultivated in bioreactor for 24 days. Production of hGH1 was 5.1 ± 0.42 µg/g dry weight (DW) for flask cultures, and 7.8 ± 0.3 µg/g DW for bioreactor, with productivity of 0.68 ± 0.05 and 1.5 ± 0.06 µg/g DW*days, respectively, indicating that the production of hGH1 was not affected by the growth rate, but might be affected by the culture system. These results demonstrate that hairy root cultures of broccoli have potential as an alternative expression system for production of hGH1, and might also be useful for production of other therapeutic proteins.

  8. Chip-based Three-dimensional Cell Culture in Perfused Micro-bioreactors

    Science.gov (United States)

    Gottwald, Eric; Lahni, Brigitte; Thiele, David; Giselbrecht, Stefan; Welle, Alexander; Weibezahn, Karl-Friedrich

    2008-01-01

    We have developed a chip-based cell culture system for the three-dimensional cultivation of cells. The chip is typically manufactured from non-biodegradable polymers, e.g., polycarbonate or polymethyl methacrylate by micro injection molding, micro hot embossing or micro thermoforming. But, it can also be manufactured from bio-degradable polymers. Its overall dimensions are 0.7 1 x 20 x 20 x 0.7 1 mm (h x w x l). The main features of the chips used are either a grid of up to 1156 cubic micro-containers (cf-chip) each the size of 120-300 x 300 x 300 μ (h x w x l) or round recesses with diameters of 300 μ and a depth of 300 μ (r-chip). The scaffold can house 10 Mio. cells in a three-dimensional configuration. For an optimal nutrient and gas supply, the chip is inserted in a bioreactor housing. The bioreactor is part of a closed steril circulation loop that, in the simplest configuration, is additionaly comprised of a roller pump and a medium reservoir with a gas supply. The bioreactor can be run in perfusion, superfusion, or even a mixed operation mode. We have successfully cultivated cell lines as well as primary cells over periods of several weeks. For rat primary liver cells we could show a preservation of organotypic functions for more than 2 weeks. For hepatocellular carcinoma cell lines we could show the induction of liver specific genes not or only slightly expressed in standard monolayer culture. The system might also be useful as a stem cell cultivation system since first differentiation experiments with stem cell lines were promising. PMID:19066592

  9. Modulation of mesenchymal stromal cell characteristics by microcarrier culture in bioreactors.

    Science.gov (United States)

    Hupfeld, Julia; Gorr, Ingo H; Schwald, Christian; Beaucamp, Nicola; Wiechmann, Kornelius; Kuentzer, Karin; Huss, Ralf; Rieger, Bernhard; Neubauer, Markus; Wegmeyer, Heike

    2014-11-01

    Mesenchymal stromal cells (MSCs) are promising candidates for cell therapy. Their therapeutic use requires extensive expansion to obtain a sufficiently high number of cells for clinical applications. State-of-the-art expansion systems, that is, primarily culture flask-based systems, are limited regarding scale-up, automation, and reproducibility. To overcome this bottleneck, microcarrier (MC)-based expansion processes have been developed. For the first time, MSCs from the perinatal sources umbilical cord (UC) and amniotic membrane (AM) were expanded on MCs. This study focuses on the comparison of flask- and Cytodex 1 MC-expanded MSCs by evaluating the influence of the expansion process on biological MSC characteristics. Furthermore, we tested the hypothesis to obtain more homogeneous MSC preparations by expanding cells on MCs in controlled large-scale bioreactors. MSCs were extensively characterized determining morphology, cell growth, surface marker expression, and functional properties such as differentiation capacity, secretion of paracrine factors, and gene expression. Based on their gene expression profile MSCs from different donors and sources clearly clustered in distinct groups solely depending on the expansion process-MC or flask culture. MC- and flask-expanded MSCs significantly differed from each other regarding surface markers and both paracrine factors and gene expression profiles. Furthermore, based on gene expression analysis, MC cultivation of MSCs in controlled bioreactor systems resulted in less heterogeneity between cells from different donors. In conclusion, MC-based MSC expansion in controlled bioreactors has the potential to reliably produce MSCs with altered characteristics and functions as compared to flask-expanded MSCs. These findings may be useful for the generation of MSCs with tailored properties for clinical applications.

  10. Cells growing in NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    For 5 days on the STS-70 mission, a bioreactor cultivated human colon cancer cells, which grew to 30 times the volume of control specimens grown on Earth. This significant result was reproduced on STS-85 which grew mature structures that more closely match what are found in tumors in humans. Shown here, clusters of cells slowly spin inside a bioreactor. On Earth, the cells continually fall through the buffer medium and never hit bottom. In space, they are naturally suspended. Rotation ensures gentle stirring so waste is removed and fresh nutrient and oxygen are supplied. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. 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 Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. 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.

  11. Cells growing in NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    For 5 days on the STS-70 mission, a bioreactor cultivated human colon cancer cells, which grew to 30 times the volume of control specimens grown on Earth. This significant result was reproduced on STS-85 which grew mature structures that more closely match what are found in tumors in humans. Shown here, clusters of cells slowly spin inside a bioreactor. On Earth, the cells continually fall through the buffer medium and never hit bottom. In space, they are naturally suspended. Rotation ensures gentle stirring so waste is removed and fresh nutrient and oxygen are supplied. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. 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 Bioreactor is rotated to provide gentle mixing of fresh and spent nutrient without inducing shear forces that would damage the cells. 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.

  12. A Novel Flow-Perfusion Bioreactor Supports 3D Dynamic Cell Culture

    Directory of Open Access Journals (Sweden)

    Alexander M. Sailon

    2009-01-01

    Full Text Available Background. Bone engineering requires thicker three-dimensional constructs than the maximum thickness supported by standard cell-culture techniques (2 mm. A flow-perfusion bioreactor was developed to provide chemotransportation to thick (6 mm scaffolds. Methods. Polyurethane scaffolds, seeded with murine preosteoblasts, were loaded into a novel bioreactor. Control scaffolds remained in static culture. Samples were harvested at days 2, 4, 6, and 8 and analyzed for cellular distribution, viability, metabolic activity, and density at the periphery and core. Results. By day 8, static scaffolds had a periphery cell density of 67%±5.0%, while in the core it was 0.3%±0.3%. Flow-perfused scaffolds demonstrated peripheral cell density of 94%±8.3% and core density of 76%±3.1% at day 8. Conclusions. Flow perfusion provides chemotransportation to thick scaffolds. This system may permit high throughput study of 3D tissues in vitro and enable prefabrication of biological constructs large enough to solve clinical problems.

  13. NASA Bioreactor Schematic

    Science.gov (United States)

    2001-01-01

    The schematic depicts the major elements and flow patterns inside the NASA Bioreactor system. Waste and fresh medium are contained in plastic bags placed side-by-side so the waste bag fills as the fresh medium bag is depleted. The compliance vessel contains a bladder to accommodate pressure transients that might damage the system. A peristolic pump moves fluid by squeezing the plastic tubing, thus avoiding potential contamination. 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.

  14. NASA Bioreactor Schematic

    Science.gov (United States)

    2001-01-01

    The schematic depicts the major elements and flow patterns inside the NASA Bioreactor system. Waste and fresh medium are contained in plastic bags placed side-by-side so the waste bag fills as the fresh medium bag is depleted. The compliance vessel contains a bladder to accommodate pressure transients that might damage the system. A peristolic pump moves fluid by squeezing the plastic tubing, thus avoiding potential contamination. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  16. Enhanced biosynthesis of withanolides by elicitation and precursor feeding in cell suspension culture of Withania somnifera (L.) Dunal in shake-flask culture and bioreactor.

    Science.gov (United States)

    Sivanandhan, Ganeshan; Selvaraj, Natesan; Ganapathi, Andy; Manickavasagam, Markandan

    2014-01-01

    The present study investigated the biosynthesis of major and minor withanolides of Withania somnifera in cell suspension culture using shake-flask culture and bioreactor by exploiting elicitation and precursor feeding strategies. Elicitors like cadmium chloride, aluminium chloride and chitosan, precursors such as cholesterol, mevalonic acid and squalene were examined. Maximum total withanolides detected [withanolide A (7606.75 mg), withanolide B (4826.05 mg), withaferin A (3732.81 mg), withanone (6538.65 mg), 12 deoxy withanstramonolide (3176.63 mg), withanoside IV (2623.21 mg) and withanoside V (2861.18 mg)] were achieved in the combined treatment of chitosan (100 mg/l) and squalene (6 mM) along with 1 mg/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose in culture at 4 h and 48 h exposure times respectively on 28th day of culture in bioreactor. We obtained higher concentrations of total withanolides in shake-flask culture (2.13-fold) as well as bioreactor (1.66-fold) when compared to control treatments. This optimized protocol can be utilized for commercial level production of withanolides from suspension culture using industrial bioreactors in a short culture period.

  17. Mechanical fatigue performance of PCL-chondroprogenitor constructs after cell culture under bioreactor mechanical stimulus.

    Science.gov (United States)

    Panadero, Juan Alberto; Sencadas, Vitor; Silva, Sonia C M; Ribeiro, Clarisse; Correia, Vitor; Gama, Francisco M; Gomez Ribelles, José Luis; Lanceros-Mendez, Senentxu

    2016-02-01

    In tissue engineering of cartilage, polymeric scaffolds are implanted in the damaged tissue and subjected to repeated compression loading cycles. The possibility of failure due to mechanical fatigue has not been properly addressed in these scaffolds. Nevertheless, the macroporous scaffold is susceptible to failure after repeated loading-unloading cycles. This is related to inherent discontinuities in the material due to the micropore structure of the macro-pore walls that act as stress concentration points. In this work, chondrogenic precursor cells have been seeded in poly-ε-caprolactone (PCL) scaffolds with fibrin and some were submitted to free swelling culture and others to cyclic loading in a bioreactor. After cell culture, all the samples were analyzed for fatigue behavior under repeated loading-unloading cycles. Moreover, some components of the extracellular matrix (ECM) were identified. No differences were observed between samples undergoing free swelling or bioreactor loading conditions, neither respect to matrix components nor to mechanical performance to fatigue. The ECM did not achieve the desired preponderance of collagen type II over collagen type I which is considered the main characteristic of hyaline cartilage ECM. However, prediction in PCL with ECM constructs was possible up to 600 cycles, an enhanced performance when compared to previous works. PCL after cell culture presents an improved fatigue resistance, despite the fact that the measured elastic modulus at the first cycle was similar to PCL with poly(vinyl alcohol) samples. This finding suggests that fatigue analysis in tissue engineering constructs can provide additional information missed with traditional mechanical measurements.

  18. Modelling wastewater treatment in a submerged anaerobic membrane bioreactor.

    Science.gov (United States)

    Spagni, Alessandro; Ferraris, Marco; Casu, Stefania

    2015-01-01

    Mathematical modelling has been widely applied to membrane bioreactor (MBRs) processes. However, to date, very few studies have reported on the application of the anaerobic digestion model N.1 (ADM1) to anaerobic membrane processes. The aim of this study was to evaluate the applicability of the ADM1 to a submerged anaerobic MBR (SAMBR) treating simulated industrial wastewater composed of cheese whey and sucrose. This study demonstrated that the biological processes involved in SAMBRs can be modelled by using the ADM1. Moreover, the results showed that very few modifications of the parameters describing the ADM1 were required to reasonably fit the experimental data. In particular, adaptation to the specific conditions of the coefficients describing the wastewater characterisation and the reduction of the hydrolysis rate of particulate carbohydrate (khyd,ch) from 0.25 d(-1) (as suggested by the ADM1 for high-rate mesophilic reactors) to 0.13 d(-1) were required to fit the experimental data.

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

    Science.gov (United States)

    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

  20. Effective enrichment of cholangiocarcinoma secretomes using the hollow fiber bioreactor culture system.

    Science.gov (United States)

    Weeraphan, Churat; Diskul-Na-Ayudthaya, Penchatr; Chiablaem, Khajeelak; Khongmanee, Amnart; Chokchaichamnankit, Daranee; Subhasitanont, Pantipa; Svasti, Jisnuson; Srisomsap, Chantragan

    2012-09-15

    The Northeastern region of Thailand is well known to have high incidence of bile duct cancer known as cholangiocarcinoma. So there is a continued need to improve diagnosis and treatment, and discovery of biomarkers for early detection of bile duct cancer should greatly improve treatment outcome for these patients. The secretome, a collection of proteins secreted from cells, is a useful source for identifying circulating biomarkers in blood secreted from cancer cells. Here a Hollow Fiber Bioreactor culture system was used for enrichment of cholangiocarcinoma secretomes, since this culture system mimics the dense three-dimensional microenvironment of the tumor found in vivo. Two-dimensional fluorescence difference gel electrophoresis using a sensitive Fluor saturation dye staining, followed by LC/MS/MS, was used to compare protein expression in the secretomes of cells cultured in the Hollow Fiber system and cells cultured in the monolayer culture system. For the first time, the 2D-patterns of cholangiocarcinoma secretomes from the two culture systems could be compared. The Hollow Fiber system improved the quality and quantity of cholangiocarcinoma secreted proteins compared to conventional monolayer system, showing less interference by cytoplasmic proteins and yielding more secreted proteins. Overall, 75 spots were analyzed by LC/MS/MS and 106 secreted proteins were identified. Two novel secreted proteins (C19orf10 and cystatin B) were found only in the Hollow Fiber system and were absent from the traditional monolayer culture system. Among the highly expressed proteins, 22 secreted soluble proteins were enriched by 5 fold in Hollow Fiber system compared to monolayer culture system. The Hollow Fiber system is therefore useful for preparing a wide range of proteins from low-abundance cell secretomes. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Monoterpenoid oxindole alkaloid production by Uncaria tomentosa (Willd) D.C. cell suspension cultures in a stirred tank bioreactor.

    Science.gov (United States)

    Trejo-Tapia, Gabriela; Cerda-García-Rojas, Carlos M; Rodríguez-Monroy, Mario; Ramos-Valdivia, Ana C

    2005-01-01

    Cell growth, monoterpenoid oxindole alkaloid (MOA) production, and morphological properties of Uncaria tomentosa cell suspension cultures in a 2-L stirred tank bioreactor were investigated. U. tomentosa (cell line green Uth-3) was able to grow in a stirred tank at an impeller tip speed of 95 cm/s (agitation speed of 400 rpm), showing a maximum biomass yield of 11.9 +/- 0.6 g DW/L and a specific growth rate of 0.102 d(-1). U. tomentosa cells growing in a stirred tank achieved maximum volumetric and specific MOA concentration (467.7 +/- 40.0 microg/L, 44.6 +/- 5.2 microg/g DW) at 16 days of culture. MOA chemical profile of cell suspension cultures growing in a stirred tank resembled that of the plant. Depending on culture time, from the total MOA produced, 37-100% was found in the medium in the bioreactor culture. MOA concentration achieved in a stirred tank was up to 10-fold higher than that obtained in Erlenmeyer flasks (agitated at 110 rpm). In a stirred tank, average area of the single cells of U. tomentosa increased up to 4-fold, and elliptical form factor increased from 1.40 to 2.55, indicating enlargement of U. tomentosa single cells. This work presents the first report of U. tomentosa green cell suspension cultures that grow and produce MOA in a stirred tank bioreactor.

  2. Large-scale functional expression of WT and truncated human adenosine A2A receptor in Pichia pastoris bioreactor cultures

    Directory of Open Access Journals (Sweden)

    Strange Philip G

    2008-10-01

    Full Text Available Abstract Background The large-scale production of G-protein coupled receptors (GPCRs for functional and structural studies remains a challenge. Recent successes have been made in the expression of a range of GPCRs using Pichia pastoris as an expression host. P. pastoris has a number of advantages over other expression systems including ability to post-translationally modify expressed proteins, relative low cost for production and ability to grow to very high cell densities. Several previous studies have described the expression of GPCRs in P. pastoris using shaker flasks, which allow culturing of small volumes (500 ml with moderate cell densities (OD600 ~15. The use of bioreactors, which allow straightforward culturing of large volumes, together with optimal control of growth parameters including pH and dissolved oxygen to maximise cell densities and expression of the target receptors, are an attractive alternative. The aim of this study was to compare the levels of expression of the human Adenosine 2A receptor (A2AR in P. pastoris under control of a methanol-inducible promoter in both flask and bioreactor cultures. Results Bioreactor cultures yielded an approximately five times increase in cell density (OD600 ~75 compared to flask cultures prior to induction and a doubling in functional expression level per mg of membrane protein, representing a significant optimisation. Furthermore, analysis of a C-terminally truncated A2AR, terminating at residue V334 yielded the highest levels (200 pmol/mg so far reported for expression of this receptor in P. pastoris. This truncated form of the receptor was also revealed to be resistant to C-terminal degradation in contrast to the WT A2AR, and therefore more suitable for further functional and structural studies. Conclusion Large-scale expression of the A2AR in P. pastoris bioreactor cultures results in significant increases in functional expression compared to traditional flask cultures.

  3. Bioreactor process parameter screening utilizing a Plackett-Burman design for a model monoclonal antibody.

    Science.gov (United States)

    Agarabi, Cyrus D; Schiel, John E; Lute, Scott C; Chavez, Brittany K; Boyne, Michael T; Brorson, Kurt A; Khan, Mansoor A; Read, Erik K

    2015-06-01

    Consistent high-quality antibody yield is a key goal for cell culture bioprocessing. This endpoint is typically achieved in commercial settings through product and process engineering of bioreactor parameters during development. When the process is complex and not optimized, small changes in composition and control may yield a finished product of less desirable quality. Therefore, changes proposed to currently validated processes usually require justification and are reported to the US FDA for approval. Recently, design-of-experiments-based approaches have been explored to rapidly and efficiently achieve this goal of optimized yield with a better understanding of product and process variables that affect a product's critical quality attributes. Here, we present a laboratory-scale model culture where we apply a Plackett-Burman screening design to parallel cultures to study the main effects of 11 process variables. This exercise allowed us to determine the relative importance of these variables and identify the most important factors to be further optimized in order to control both desirable and undesirable glycan profiles. We found engineering changes relating to culture temperature and nonessential amino acid supplementation significantly impacted glycan profiles associated with fucosylation, β-galactosylation, and sialylation. All of these are important for monoclonal antibody product quality.

  4. Transgenic rabbits as therapeutic protein bioreactors and human disease models.

    Science.gov (United States)

    Fan, Jianglin; Watanabe, Teruo

    2003-09-01

    Genetically modified laboratory animals provide a powerful approach for studying gene expression and regulation and allow one to directly examine structure-function and cause-and-effect relationships in pathophysiological processes. Today, transgenic mice are available as a research tool in almost every research institution. On the other hand, the development of a relatively large mammalian transgenic model, transgenic rabbits, has provided unprecedented opportunities for investigators to study the mechanisms of human diseases and has also provided an alternative way to produce therapeutic proteins to treat human diseases. Transgenic rabbits expressing human genes have been used as a model for cardiovascular disease, AIDS, and cancer research. The recombinant proteins can be produced from the milk of transgenic rabbits not only at lower cost but also on a relatively large scale. One of the most promising and attractive recombinant proteins derived from transgenic rabbit milk, human alpha-glucosidase, has been successfully used to treat the patients who are genetically deficient in this enzyme. Although the pronuclear microinjection is still the major and most popular method for the creation of transgenic rabbits, recent progress in gene targeting and animal cloning has opened new avenues that should make it possible to produce transgenic rabbits by somatic cell nuclear transfer in the future. Based on a computer-assisted search of the studies of transgenic rabbits published in the English literature here, we introduce to the reader the achievements made thus far with transgenic rabbits, with emphasis on the application of these rabbits as human disease models and live bioreactors for producing human therapeutic proteins and on the recent progress in cloned rabbits.

  5. 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.

  6. Tissue grown in space in NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    For 5 days on the STS-70 mission, a bioreactor cultivated human colon cancer cells, such as the culture section shown here, which grew to 30 times the volume of control specimens grown on Earth. This significant result was reproduced on STS-85 which grew mature structures that more closely match what are found in tumors in humans. The two white circles within the tumor are part of a plastic lattice that helped the cells associate. 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). The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. 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. 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.

  7. 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

  8. A photosynthetic rotating annular bioreactor (Taylor-Couette type flow) for phototrophic biofilm cultures.

    Science.gov (United States)

    Paule, A; Lauga, B; Ten-Hage, L; Morchain, J; Duran, R; Paul, E; Rols, J L

    2011-11-15

    In their natural environment, the structure and functioning of microbial communities from river phototrophic biofilms are driven by biotic and abiotic factors. An understanding of the mechanisms that mediate the community structure, its dynamics and the biological succession processes during phototrophic biofilm development can be gained using laboratory-scale systems operating with controlled parameters. For this purpose, we present the design and description of a new prototype of a rotating annular bioreactor (RAB) (Taylor-Couette type flow, liquid working volume of 5.04 L) specifically adapted for the cultivation and investigation of phototrophic biofilms. The innovation lies in the presence of a modular source of light inside of the system, with the biofilm colonization and development taking place on the stationary outer cylinder (onto 32 removable polyethylene plates). The biofilm cultures were investigated under controlled turbulent flowing conditions and nutrients were provided using a synthetic medium (tap water supplemented with nitrate, phosphate and silica) to favour the biofilm growth. The hydrodynamic features of the water flow were characterized using a tracer method, showing behaviour corresponding to a completely mixed reactor. Shear stress forces on the surface of plates were also quantified by computer simulations and correlated with the rotational speed of the inner cylinder. Two phototrophic biofilm development experiments were performed for periods of 6.7 and 7 weeks with different inoculation procedures and illumination intensities. For both experiments, biofilm biomasses exhibited linear growth kinetics and produced 4.2 and 2.4 mg cm(-)² of ash-free dry matter. Algal and bacterial community structures were assessed by microscopy and T-RFLP, respectively, and the two experiments were different but revealed similar temporal dynamics. Our study confirmed the performance and multipurpose nature of such an innovative photosynthetic bioreactor

  9. High-density mammalian cell cultures in stirred-tank bioreactor without external pH control.

    Science.gov (United States)

    Xu, Sen; Chen, Hao

    2016-08-10

    Maintaining desired pH is a necessity for optimal cell growth and protein production. It is typically achieved through a two-sided pH control loop on the bioreactor controller. Here we investigated cell culture processes with minimum or no pH control and demonstrated that high-density mammalian cell cultures could be maintained for long-term protein production without pH control. The intrinsic interactions between pCO2, lactate, and pH were leveraged to maintain culture pH. Fed-batch cultures at the same lower pH limit of 6.75 but different upper pH limits (7.05, 7.30, 7.45, 7.65) were evaluated in the 3L bioreactors and comparable results were obtained. Neither CO2 sparging nor base addition was required to control pH in the pH range of 6.75-7.65. The impact of sparger configurations (drilled hole sparger vs. frit sparger) and scales (3L vs. 200L) on CO2 accumulation and culture pH was also demonstrated. The same principle was applied in two perfusion cultures with steady state cell densities at 42.5±3.3 or 68.3±6.0×10(6)cells/mL with low cell specific perfusion rates (15±2 to 23±3pL/cell/day), achieving up to 1.9±0.1g/L/day bioreactor productivity. Culture pH level in the 3L perfusion bioreactors was steadily maintained by controlling the residual lactate and pCO2 levels without the requirement of external pH control for up to 40days with consistent productivity and product quality. Furthermore, culture pH could be potentially modulated via adjusting residual glucose levels and CO2 stripping capability in perfusion cultures. To the best of our knowledge, this is the first time a systematic study was performed to evaluate the long-term cell cultivation and protein production in stirred-tank bioreactors without external pH control.

  10. Modelling and characterization of an airlift-loop bioreactor.

    NARCIS (Netherlands)

    Verlaan, P.

    1987-01-01

    An airlift-loop reactor is a bioreactor for aerobic biotechnological processes. The special feature of the ALR is the recirculation of the liquid through a downcomer connecting the top and the bottom of the main bubbling section. Due to the high circulation-flow rate, efficient mixing and oxygen tra

  11. Simulating woodchip bioreactor performance using a dual-porosity model

    Science.gov (United States)

    Nitrate in the Nation's surface waters has been a persistent health and ecological problem. The major source of nitrate is tile drainage from agricultural row crops. Denitrification bioreactors have been shown to be effective in removing much of the nitrate from tile drains. While we understand i...

  12. Hollow fiber bioreactor technology for tissue engineering applications.

    Science.gov (United States)

    Eghbali, Hadis; Nava, Michele M; Mohebbi-Kalhori, Davod; Raimondi, Manuela T

    2016-01-01

    Hollow fiber bioreactors are the focus of scientific research aiming to mimic physiological vascular networks and engineer organs and tissues in vitro. The reason for this lies in the interesting features of this bioreactor type, including excellent mass transport properties. Indeed, hollow fiber bioreactors allow limitations to be overcome in nutrient transport by diffusion, which is often an obstacle to engineer sizable constructs in vitro. This work reviews the existing literature relevant to hollow fiber bioreactors in organ and tissue engineering applications. To this purpose, we first classify the hollow fiber bioreactors into 2 categories: cylindrical and rectangular. For each category, we summarize their main applications both at the tissue and at the organ level, focusing on experimental models and computational studies as predictive tools for designing innovative, dynamic culture systems. Finally, we discuss future perspectives on hollow fiber bioreactors as in vitro models for tissue and organ engineering applications.

  13. A novel automated bioreactor for scalable process optimisation of haematopoietic stem cell culture.

    Science.gov (United States)

    Ratcliffe, E; Glen, K E; Workman, V L; Stacey, A J; Thomas, R J

    2012-10-31

    Proliferation and differentiation of haematopoietic stem cells (HSCs) from umbilical cord blood at large scale will potentially underpin production of a number of therapeutic cellular products in development, including erythrocytes and platelets. However, to achieve production processes that are scalable and optimised for cost and quality, scaled down development platforms that can define process parameter tolerances and consequent manufacturing controls are essential. We have demonstrated the potential of a new, automated, 24×15 mL replicate suspension bioreactor system, with online monitoring and control, to develop an HSC proliferation and differentiation process for erythroid committed cells (CD71(+), CD235a(+)). Cell proliferation was relatively robust to cell density and oxygen levels and reached up to 6 population doublings over 10 days. The maximum suspension culture density for a 48 h total media exchange protocol was established to be in the order of 10(7)cells/mL. This system will be valuable for the further HSC suspension culture cost reduction and optimisation necessary before the application of conventional stirred tank technology to scaled manufacture of HSC derived products.

  14. Glycosylation analysis of an aggregated antibody produced by Chinese hamster ovary cells in bioreactor culture.

    Science.gov (United States)

    Onitsuka, Masayoshi; Kawaguchi, Akira; Asano, Ryutaro; Kumagai, Izumi; Honda, Kohsuke; Ohtake, Hisao; Omasa, Takeshi

    2014-05-01

    N-Glycosylation of therapeutic antibodies contributes not only to their biological function, but also to their stability and tendency to aggregate. Here, we investigated the impact of the glycosylation status of an aggregated antibody that accumulated during the bioreactor culture of Chinese hamster ovary cells. High-performance liquid chromatography analysis showed that there was no apparent difference in the glycosylation patterns of monomeric, dimeric, and large aggregated forms of the antibody. In contrast, lectin binding assays, which enable the total amounts of specific sugar residues to be detected, showed that both galactose and fucose residues in dimers and large aggregates were reduced to 70-80% of the amount in monomers. These results strongly suggest that the lack of N-linked oligosaccharides, a result of deglycosylation or aglycosylation, occurred in a proportion of the dimeric and large aggregated components. The present study demonstrates that glycosylation heterogeneities are a potential cause of antibody aggregation in cell culture of Chinese hamster ovary cells, and that the lack of N-glycosylation promotes the formation of dimers and finally results in large aggregates.

  15. Bacterial Concentration and Diversity within Repetitive Aliquots Collected from Replicate Continuous-Flow Bioreactor Cultures.

    Science.gov (United States)

    Crippen, Tawni L; Sheffield, Cynthia L; Andrews, Kathleen; Bongaerts, Roy; Nisbet, David J

    2008-01-01

    The aim of this study was to determine the reproducibility of small volume repeat sampling from replicate bioreactors with stabilized continuous-flow chicken cecal bacterial communities. Bacterial concentration and diversity were analyzed by phenotypic, biochemical and ribotype analysis. Significant differences in concentrations and variations in diversity were found in replicate bioreactors.

  16. Recombinant Escherichia coli strains with inducible Campylobacter jejuni single domain hemoglobin CHb expression exhibited improved cell growth in bioreactor culture.

    Directory of Open Access Journals (Sweden)

    Li Xu

    Full Text Available Maintaining an appropriate concentration of dissolved oxygen in aqueous solution is critical for efficient operation of a bioreactor, requiring sophisticated engineering design and a system of regulation to maximize oxygen transfer from the injected air bubbles to the cells. Bacterial hemoglobins are oxygen-binding proteins that transfer oxygen from the environment to metabolic processes and allow bacteria to grow even under microaerophilic conditions. To improve the oxygen utilization efficiency of cells and overcome the oxygen shortage in bioreactors, the gene coding for the Campylobacter jejuni single domain hemoglobin (CHb gene was artificially synthesized and functionally expressed under the control of inducible expression promoters PT7 and Pvgh in Escherichia coli. The effects of the recombinants PT7-CHb and Pvgh-CHb on cell growth were evaluated in aerobic shake flasks, anaerobic capped bottles and a 5-L bioreactor, and a pronounced improvement in cell biomass was observed for CHb-expressing cells. To determine the growth curves, CHb gene expression, and CHb oxygen-binding capacity of specific recombinants with different promoters, we determined the time course of CHb gene expression in the two recombinants by semi-quantitative RT-PCR and CO differential spectrum assays. Based on the growth patterns of the two recombinants in the bioreactor, we proposed different recombinant types with optimal performance under specific culture conditions.

  17. A Multi-Paradigm Modeling Framework to Simulate Dynamic Reciprocity in a Bioreactor

    Science.gov (United States)

    Kaul, Himanshu; Cui, Zhanfeng; Ventikos, Yiannis

    2013-01-01

    Despite numerous technology advances, bioreactors are still mostly utilized as functional black-boxes where trial and error eventually leads to the desirable cellular outcome. Investigators have applied various computational approaches to understand the impact the internal dynamics of such devices has on overall cell growth, but such models cannot provide a comprehensive perspective regarding the system dynamics, due to limitations inherent to the underlying approaches. In this study, a novel multi-paradigm modeling platform capable of simulating the dynamic bidirectional relationship between cells and their microenvironment is presented. Designing the modeling platform entailed combining and coupling fully an agent-based modeling platform with a transport phenomena computational modeling framework. To demonstrate capability, the platform was used to study the impact of bioreactor parameters on the overall cell population behavior and vice versa. In order to achieve this, virtual bioreactors were constructed and seeded. The virtual cells, guided by a set of rules involving the simulated mass transport inside the bioreactor, as well as cell-related probabilistic parameters, were capable of displaying an array of behaviors such as proliferation, migration, chemotaxis and apoptosis. In this way the platform was shown to capture not only the impact of bioreactor transport processes on cellular behavior but also the influence that cellular activity wields on that very same local mass transport, thereby influencing overall cell growth. The platform was validated by simulating cellular chemotaxis in a virtual direct visualization chamber and comparing the simulation with its experimental analogue. The results presented in this paper are in agreement with published models of similar flavor. The modeling platform can be used as a concept selection tool to optimize bioreactor design specifications. PMID:23555740

  18. A multi-paradigm modeling framework to simulate dynamic reciprocity in a bioreactor.

    Directory of Open Access Journals (Sweden)

    Himanshu Kaul

    Full Text Available Despite numerous technology advances, bioreactors are still mostly utilized as functional black-boxes where trial and error eventually leads to the desirable cellular outcome. Investigators have applied various computational approaches to understand the impact the internal dynamics of such devices has on overall cell growth, but such models cannot provide a comprehensive perspective regarding the system dynamics, due to limitations inherent to the underlying approaches. In this study, a novel multi-paradigm modeling platform capable of simulating the dynamic bidirectional relationship between cells and their microenvironment is presented. Designing the modeling platform entailed combining and coupling fully an agent-based modeling platform with a transport phenomena computational modeling framework. To demonstrate capability, the platform was used to study the impact of bioreactor parameters on the overall cell population behavior and vice versa. In order to achieve this, virtual bioreactors were constructed and seeded. The virtual cells, guided by a set of rules involving the simulated mass transport inside the bioreactor, as well as cell-related probabilistic parameters, were capable of displaying an array of behaviors such as proliferation, migration, chemotaxis and apoptosis. In this way the platform was shown to capture not only the impact of bioreactor transport processes on cellular behavior but also the influence that cellular activity wields on that very same local mass transport, thereby influencing overall cell growth. The platform was validated by simulating cellular chemotaxis in a virtual direct visualization chamber and comparing the simulation with its experimental analogue. The results presented in this paper are in agreement with published models of similar flavor. The modeling platform can be used as a concept selection tool to optimize bioreactor design specifications.

  19. Pilot-scale culture of Hypericum perforatum L. adventitious roots in airlift bioreactors for the production of bioactive compounds.

    Science.gov (United States)

    Cui, Xi-Hua; Murthy, Hosakatte Niranjana; Paek, Kee-Yoeup

    2014-09-01

    Hypericum perforatum L. (St. John's Wort) is an important medicinal plant which is widely used in the treatment for depression and irritable bowel syndrome. It is also used as a dietary supplement. Major bioactive phytochemicals of H. perforatum are phenolics and flavonoids. Quality of these phytochemicals is dramatically influenced by environmental and biological factors in the field grown plants. As an alternative, we have developed adventitious root cultures in large-scale bioreactors for the production of useful phytochemicals. Adventitious roots of H. perforatum were cultured in 500 l pilot-scale airlift bioreactors using half-strength Murashige and Skoog medium with an ammonium and nitrate ratio of 5:25 mM and supplemented with 1.0 mg l(-1) indole butyric acid, 0.1 mg l(-1) kinetin, and 3 % sucrose for the production of bioactive phenolics and flavonoids. Then 4.6 and 6.3 kg dry biomass were realized in the 500 l each of drum-type and balloon-type bioreactors, respectively. Accumulation of 66.9 mg g(-1) DW of total phenolics, 48.6 mg g(-1) DW of total flavonoids, 1.3 mg g(-1) DW of chlorogenic acid, 0.01 mg g(-1) DW of hyperin, 0.04 mg g(-1) DW of hypericin, and 0.01 mg g(-1) DW of quercetin could be achieved with adventitious roots cultured in 500 l balloon-type airlift bioreactors. Our findings demonstrate the possibilities of using H. perforatum adventitious root cultures for the production of useful phytochemicals to meet the demand of pharmaceutical and food industry.

  20. Quality of mixing in a stired bioreactor used for animal cells culture: heterogeneities in a lab scale bioreactor and evolution of mixing time with scale up

    Directory of Open Access Journals (Sweden)

    Collignon, ML.

    2010-01-01

    Full Text Available Animal cells are industrially cultivated inside stirred bioreactors to produce proteinic compounds. Due to the use of mild agitation conditions in order to limit mechanical constraints, the homogeneity of the culture medium can be far from perfect. This study has therefore two objectives: the global characterization of the mixing via the mixing time and the local description of concentration fields. The mixing time is measured by conductimetry inside 20 l, 80 l, 600 l tanks. The Grenville correlation is adjusted on these experimental measurements to improve the prediction of the mixing time during the scale-up of the process. The concentration fields are visualized by the Planar Laser Induced Fluorescence (P.L.I.F. technique in the 20 l tank. This part of the study is focused on the time evolution of the maximum value of the tracer concentration inside measurement planes and of the numerical distribution of theses concentration fields.

  1. 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.

  2. Inter-microcarrier transfer and phenotypic stability of stem cell-derived Schwann cells in stirred suspension bioreactor culture.

    Science.gov (United States)

    Shakhbazau, Antos; Mirfeizi, Leila; Walsh, Tylor; Wobma, Holly M; Kumar, Ranjan; Singh, Bhagat; Kallos, Michael S; Midha, Rajiv

    2016-02-01

    Emerging bioreactor technologies offer an effective way for scaled-up production of large numbers of cells for cell therapy applications. One of the clinical paradigms where cell therapy can be an asset is restorative neurosciences. Nerve repair can benefit from the injections of stem cells and/or Schwann cells, acting as a source for axon myelination, myelin debris clearance, and trophic support. We have adapted microcarrier-based suspension bioreactor culture for Schwann cells (SCs) differentiated from a new stem cell source - skin-derived precursors (SKPs). SKP-derived SCs attach and grow on different types of microcarriers in both static and stirred culture, with Cytodex 3 and CultiSpher-S found most effective. Inter-microcarrier migration of SKP-SCs represents a key mechanism for rapid expansion and colonization in stirred suspension culture. We have shown that microcarrier-expanded SKP-SCs cells express Schwann cell markers p75-NTR, GFAP and S100 and retain their key ability to myelinate axons both in vitro and in vivo. Scaled-up microcarrier-based production of SKP-SCs in suspension bioreactors appears feasible for timely generation of sufficient cell numbers for nerve repair strategies.

  3. Production of ethanol directly from potato starch by mixed culture of Saccharomyces cerevisiae and Aspergillus niger using electrochemical bioreactor.

    Science.gov (United States)

    Jeon, Bo Young; Kim, Dae Hee; Na, Byung Kwan; Ahn, Dae Hee; Park, Doo Hyun

    2008-03-01

    When cultivated aerobically, Aspergillus niger hyphae produced extracellular glucoamylase, which catalyzes the saccharification of unliquified potato starch into glucose, but not when grown under anaerobic conditions. The Km and Vmax of the extracellular glucoamylase were 652.3 mg starch l-1 and 253.3 mg glucose l-1 min-1, respectively. In mixed culture of A. niger and Saccharomyces cerevisiae, oxygen had a negative influence on the alcohol fermentation of yeast, but activated fungal growth. Therefore, oxygen is a critical factor for ethanol production in the mixed culture, and its generation through electrolysis of water in an electrochemical bioreactor needs to be optimized for ethanol production from starch by coculture of fungal hyphae and yeast cells. By applying pulsed electric fields (PEF) into the electrochemical bioreactor, ethanol production from starch improved significantly: Ethanol produced from 50 g potato starch l-1 by a mixed culture of A. niger and S. cerevisiae was about 5 g l-1 in a conventional bioreactor, but was 9 g l-1 in 5 volts of PEF and about 19 g l-1 in 4 volts of PEF for 5 days.

  4. 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; Hewitt, Christopher J

    2016-03-01

    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.

  5. Modelling the damage potential of fluid flows for animal cells undergoing cultivation in bioreactors.

    Science.gov (United States)

    Stanford Keen, Giles

    1996-11-01

    Mechanical disruption and injury sustained by animal cells undergoing cultivation in bioreactors is an important problem in biotechnology. Damage to cells is thought to be caused primarily by bubbles bursting at the free surface of the culture medium. Here we present computational studies applying a mathematical model for the cell damage rates experienced by cells in laminar flow. Two fluid dynamical systems are considered - namely a converging channel and a single bursting bubble. The flows are calculated using a fourth-order finite difference technique on a stretched grid, and a boundary integral method respectively. It is possible to obtain an estimate for the number of cells in a particular population which are likely to be disrupted by the forces they experience in the flow. This is done by calculating the maximum rate of strain experienced by fluid particles, and combining this with experimental data on the strength and size of cells, obtained by micromanipulation techniques. The resulting information is then used together with the cell damage model to produce a cell damage prediction. The computational results are compared with experimental measurements of cell death, to validate the model for cell damage.

  6. A mathematical model and computational framework for three-dimensional chondrocyte cell growth in a porous tissue scaffold placed inside a bi-directional flow perfusion bioreactor.

    Science.gov (United States)

    Shakhawath Hossain, Md; Bergstrom, D J; Chen, X B

    2015-12-01

    The in vitro chondrocyte cell culture for cartilage tissue regeneration in a perfusion bioreactor is a complex process. Mathematical modeling and computational simulation can provide important insights into the culture process, which would be helpful for selecting culture conditions to improve the quality of the developed tissue constructs. However, simulation of the cell culture process is a challenging task due to the complicated interaction between the cells and local fluid flow and nutrient transport inside the complex porous scaffolds. In this study, a mathematical model and computational framework has been developed to simulate the three-dimensional (3D) cell growth in a porous scaffold placed inside a bi-directional flow perfusion bioreactor. The model was developed by taking into account the two-way coupling between the cell growth and local flow field and associated glucose concentration, and then used to perform a resolved-scale simulation based on the lattice Boltzmann method (LBM). The simulation predicts the local shear stress, glucose concentration, and 3D cell growth inside the porous scaffold for a period of 30 days of cell culture. The predicted cell growth rate was in good overall agreement with the experimental results available in the literature. This study demonstrates that the bi-directional flow perfusion culture system can enhance the homogeneity of the cell growth inside the scaffold. The model and computational framework developed is capable of providing significant insight into the culture process, thus providing a powerful tool for the design and optimization of the cell culture process.

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

    Science.gov (United States)

    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.

  8. Use of a Three-Dimensional Reactive Solute Transport Model for Evaluation of Bioreactor Placement in Stream Restoration.

    Science.gov (United States)

    Cui, Zhengtao; Welty, Claire; Gold, Arthur J; Groffman, Peter M; Kaushal, Sujay S; Miller, Andrew J

    2016-05-01

    A three-dimensional groundwater flow and multispecies reactive transport model was used to strategically design placement of bioreactors in the subsurface to achieve maximum removal of nitrate along restored stream reaches. Two hypothetical stream restoration scenarios were evaluated over stream reaches of 40 and 94 m: a step-pool scenario and a channel re-meandering scenario. For the step-pool scenario, bioreactors were placed at locations where mass fluxes of groundwater and nitrate were highest. Bioreactors installed over 50% of the total channel length of a step-pool scenario (located to intercept maximum groundwater and nitrate mass flux) removed nitrate-N entering the channel at a rate of 36.5 kg N yr (100 g N d), achieving about 65% of the removal of a whole-length bioreactor. Bioreactor placement for the re-meandering scenario was designed using a criterion of either highest nitrate mass flux or highest groundwater flux, but not both, because they did not occur together. Bioreactors installed at maximum nitrate flux locations (53% of the total channel length) on the western bank removed nitrate-N entering the channel at 62.0 kg N yr (170 g N d), achieving 85% of nitrate-N removal of whole-length bioreactors for the re-meandering scenario. Bioreactors installed at maximum groundwater flux locations on the western bank along approximately 40% of the re-meandering channel achieved about 65% of nitrate removal of whole-length bioreactors. Placing bioreactors at maximum nitrate flux locations improved denitrification efficiency. Due to low groundwater velocities, bioreactor nitrate-N removal was found to be nitrate limited for all scenarios.

  9. [Effect evaluation of three cell culture models].

    Science.gov (United States)

    Wang, Aiguo; Xia, Tao; Yuan, Jing; Chen, Xuemin

    2003-11-01

    Primary rat hepatocytes were cultured using three kinds of models in vitro and the enzyme leakage, albumin secretion, and cytochrome P450 1A (CYP 1A) activity were observed. The results showed that the level of LDH in the medium decreased over time in the period of culture. However, on 5 days, LDH showed a significant increase in monolayer culture (MC) while after 8 days LDH was not detected in sandwich culture (SC). The levels of AST and ALT in the medium did not change significantly over the investigated time. The basic CYP 1A activity gradually decreased with time in MC and SC. The decline of CYP 1A in rat hepatocytes was faster in MC than that in SC. This effect was partially reversed by using cytochrome P450 (CYP450) inducers such as omeprazol and 3-methylcholanthrene (3-MC) and the CYP 1A induction was always higher in MC than that in SC. Basic CYP 1A activity in bioreactor was keeped over 2 weeks and the highest albumin production was observed in bioreactor, and next were SC and MC. In conclusion, our results clearly indicated that there have some advantages and disadvantages in each of models in which can address different questions in metabolism of toxicants and drugs.

  10. A hydrodynamics-reaction kinetics coupled model for evaluating bioreactors derived from CFD simulation.

    Science.gov (United States)

    Wang, Xu; Ding, Jie; Guo, Wan-Qian; Ren, Nan-Qi

    2010-12-01

    Investigating how a bioreactor functions is a necessary precursor for successful reactor design and operation. Traditional methods used to investigate flow-field cannot meet this challenge accurately and economically. Hydrodynamics model can solve this problem, but to understand a bioreactor in sufficient depth, it is often insufficient. In this paper, a coupled hydrodynamics-reaction kinetics model was formulated from computational fluid dynamics (CFD) code to simulate a gas-liquid-solid three-phase biotreatment system for the first time. The hydrodynamics model is used to formulate prediction of the flow field and the reaction kinetics model then portrays the reaction conversion process. The coupled model is verified and used to simulate the behavior of an expanded granular sludge bed (EGSB) reactor for biohydrogen production. The flow patterns were visualized and analyzed. The coupled model also demonstrates a qualitative relationship between hydrodynamics and biohydrogen production. The advantages and limitations of applying this coupled model are discussed.

  11. Colon tumor cells grown in NASA Bioreactor

    Science.gov (United States)

    2001-01-01

    These photos compare the results of colon carcinoma cells grown in a NASA Bioreactor flown on the STS-70 Space Shuttle in 1995 flight and ground control experiments. The cells grown in microgravity (left) have aggregated to form masses that are larger and more similar to tissue found in the body than the cells cultured on the ground (right). The principal investigator is Milburn Jessup of the University of Texas M. D. Anderson Cancer Center. The NASA Bioreactor provides a low turbulence culture environment which promotes the formation of large, three-dimensional cell clusters. 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. 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. Cell constructs grown in a rotating bioreactor on Earth (left) eventually become too large to stay suspended in the nutrient media. In the microgravity of orbit, the cells stay suspended. Rotation then is needed for gentle stirring to replenish the media around the cells. 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). Credit: NASA and University of Texas M. D. Anderson Cancer Center.

  12. Three-dimensional Expansion: In Suspension Culture of SD Rat's Osteoblasts in a Rotating Wall Vessel Bioreactor

    Institute of Scientific and Technical Information of China (English)

    KE-DONG SONG; TIAN-QING LIU; XIANG-QIN LI; ZHAN-FENG CUI; XIANG-YU SUN; XUE-HU MA

    2007-01-01

    Objective To study large-scale expansion of SD (Sprague-Dawley) rat's osteoblasts in suspension culture in a rotating wall vessel bioreactor (RWVB). Methods The bioreactor rotation speeds were adjusted in the range of 0 to 20 rpm,which could provide low shear on the microcarriers around 1 dyn/cm2. The cells were isolated via sequential digestions of neonatal (less than 3 days old) SD rat calvaria. After the primary culture and several passages, the cells were seeded onto the microcarriers and cultivated in T-flask, spinner flask and RWVB respectively. During the culture period, the cells were counted and observed under the inverted microscope for morphology every 12 h. After 7 days, the cells were evaluated with scanning electron microscope (SEM) for histological examination of the aggregates. Also, the hematoxylin-eosin (HE) staining and alkaline phosphatase (ALP) staining were performed. Moreover, von-Kossa staining and Alizarin Red S staining were carried out for mineralized nodule formation. Results The results showed that in RWVB, the cells could be expanded by more than ten times and they presented better morphology and vitality and stronger ability to form bones. Conclusions The developed RWVB can provide the culture environment with a relatively low shear force and necessary three-dimensional (3D)interactions among cells and is suitable for osteopath expansion in vitro.

  13. Kinetic modelling and microbial community assessment of anaerobic biphasic fixed film bioreactor treating distillery spent wash.

    Science.gov (United States)

    Acharya, Bhavik K; Pathak, Hilor; Mohana, Sarayu; Shouche, Yogesh; Singh, Vasdev; Madamwar, Datta

    2011-08-01

    Anaerobic digestion, microbial community structure and kinetics were studied in a biphasic continuously fed, upflow anaerobic fixed film reactor treating high strength distillery wastewater. Treatment efficiency of the bioreactor was investigated at different hydraulic retention times (HRT) and organic loading rates (OLR 5-20 kg COD m⁻³ d⁻¹). Applying the modified Stover-Kincannon model to the reactor, the maximum removal rate constant (U(max)) and saturation value constant (K(B)) were found to be 2 kg m⁻³ d⁻¹ and 1.69 kg m⁻³ d⁻¹ respectively. Bacterial community structures of acidogenic and methanogenic reactors were assessed using culture-independent analyses. Sequencing of 16S rRNA genes exhibited a total of 123 distinct operational taxonomic units (OTUs) comprising 49 from acidogenic reactor and 74 (28 of eubacteria and 46 of archaea) from methanogenic reactor. The findings reveal the role of Lactobacillus sp. (Firmicutes) as dominant acid producing organisms in acidogenic reactor and Methanoculleus sp. (Euryarchaeotes) as foremost methanogens in methanogenic reactor.

  14. 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.

  15. Computer modeling movement of biomass in the bioreactors with bubbling mixing

    Science.gov (United States)

    Kuschev, L. A.; Suslov, D. Yu; Alifanova, A. I.

    2017-01-01

    Recently in the Russian Federation there is an observation of the development of biogas technologies which are used in organic waste conversion of agricultural enterprises, consequently improving the ecological environment. To intensify the process and effective outstanding performance of the acquisition of biogas the application of systems of mixing of bubbling is used. In the case of bubbling mixing of biomass in the bioreactor two-phase portions consisting of biomass and bubbles of gas are formed. The bioreactor computer model with bubble pipeline has been made in a vertical spiral form forming a cone type turned upside down. With the help of computing program of OpenFVM-Flow, an evaluation experiment was conducted to determine the key technological parameters of process of bubbling mixing and to get a visual picture of biomass flows distribution in the bioreactor. For the experimental bioreactor the following equation of V=190 l, speed level, the biomass circulation, and the time of a single cycle of uax =0,029 m/s; QC =0,00087 m3/s, Δtbm .=159 s. In future, we plan to conduct a series of theoretical and experimental researches into the mixing frequency influence on the biogas acquisition process effectiveness.

  16. Effect of ambient light on monoclonal antibody product quality during small-scale mammalian cell culture process in clear glass bioreactors.

    Science.gov (United States)

    Mallaney, Mary; Wang, Szu-Han; Sreedhara, Alavattam

    2014-01-01

    During a small-scale cell culture process producing a monoclonal antibody, a larger than expected difference was observed in the charge variants profile of the harvested cell culture fluid (HCCF) between the 2 L and larger scales (e.g., 400 L and 12 kL). Small-scale studies performed at the 2 L scale consistently showed an increase in acidic species when compared with the material made at larger scale. Since the 2 L bioreactors were made of clear transparent glass while the larger scale reactors are made of stainless steel, the effect of ambient laboratory light on cell culture process in 2 L bioreactors as well as handling the HCCF was carefully evaluated. Photoreactions in the 2 L glass bioreactors including light mediated increase in acidic variants in HCCF and formulation buffers were identified and carefully analyzed. While the acidic variants comprised of a mixture of sialylated, reduced disulfide, crosslinked (nonreducible), glycated, and deamidated forms, an increase in the nonreducible forms, deamidation and Met oxidation was predominantly observed under light stress. The monoclonal antibody produced in glass bioreactors that were protected from light behaved similar to the one produced in the larger scale. Our data clearly indicate that care should be taken when glass bioreactors are used in cell culture studies during monoclonal antibody production.

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

    Directory of Open Access Journals (Sweden)

    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.

  18. Scale-up of human embryonic stem cell culture using a hollow fibre bioreactor.

    Science.gov (United States)

    Roberts, Iwan; Baila, Stefano; Rice, R Brent; Janssens, Michiel Etienne; Nguyen, Kim; Moens, Nathalie; Ruban, Ludmila; Hernandez, Diana; Coffey, Pete; Mason, Chris

    2012-12-01

    The commercialisation of human embryonic stem cell derived cell therapies for large patient populations is reliant on both minimising expensive and variable manual-handling methods whilst realising economies of scale. The Quantum Cell Expansion System, a hollow fibre bioreactor (Terumo BCT), was used in a pilot study to expand 60 million human embryonic stem cells to 708 million cells. Further improvements can be expected with optimisation of media flow rates throughout the run to better control the cellular microenvironment. High levels of pluripotency marker expression were maintained on the bioreactor, with 97.7 % of cells expressing SSEA-4 when harvested.

  19. On-line monitoring of respiration in recombinant-baculovirus infected and uninfected insect cell bioreactor cultures.

    Science.gov (United States)

    Kamen, A A; Bédard, C; Tom, R; Perret, S; Jardin, B

    1996-04-05

    Respiration rates in Spodoptera frugiperda (Sf-9) cell bioreactor cultures were successfully measured on-line using two methods: The O(2) uptake rate (OUR) was determined using gas phase pO(2) values imposed by a dissolved oxygen controller and the CO(2) evolution rate (CER) was measured using an infrared detector. The measurement methods were accurate, reliable, and relatively inexpensive. The CER was routinely determined in bioreactor cultures used for the production of several recombinant proteins. Simple linear relationships between viable cell densities and both OUR and CER in exponentially growing cultures were used to predict viable cell density. Respiration measurements were also used to follow the progress of baculoviral infections in Sf-9 cultures. Infection led to increases in volumetric and per-cell respiration rates. The relationships between respiration and several other culture parameters, including viable cell density, cell protein, cell volume, glucose consumption, lactate production, viral titer, and recombinant beta-galactosidase accumulation, were examined. The extent of the increase in CER following infection and the time postinfection at which maximum CER was attained were negatively correlated with the multiplicity of infection (MOI) at multiplicities below the level required to infect all the cells in a culture. Delays in the respiration peak related to the MOI employed were correlated with delays in the peak in recombinant protein accumulation. DO levels in the range 5-100% did not exert any major effects on viable cell densities, CER, or product titer in cultures infected with a baculovirus expressing recombinant beta-galactosidase.

  20. 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.

  1. Heart tissue grown in NASA Bioreactor

    Science.gov (United States)

    2001-01-01

    Lisa Freed and Gordana Vunjak-Novakovic, both of the Massachusetts Institute of Technology (MIT), have taken the first steps toward engineering heart muscle tissue that could one day be used to patch damaged human hearts. Cells isolated from very young animals are attached to a three-dimensional polymer scaffold, then placed in a NASA bioreactor. The cells do not divide, but after about a week start to cornect to form a functional piece of tissue. Functionally connected heart cells that are capable of transmitting electrical signals are the goal for Freed and Vunjak-Novakovic. Electrophysiological recordings of engineered tissue show spontaneous contractions at a rate of 70 beats per minute (a), and paced contractions at rates of 80, 150, and 200 beats per minute respectively (b, c, and d). 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. 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. 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). Credit: NASA and MIT.

  2. Serum-free culture of primary human hepatocytes in a miniaturized hollow-fibre membrane bioreactor for pharmacological in vitro studies.

    Science.gov (United States)

    Lübberstedt, Marc; Müller-Vieira, Ursula; Biemel, Klaus M; Darnell, Malin; Hoffmann, Stefan A; Knöspel, Fanny; Wönne, Eva C; Knobeloch, Daniel; Nüssler, Andreas K; Gerlach, Jörg C; Andersson, Tommy B; Zeilinger, Katrin

    2015-09-01

    Primary human hepatocytes represent an important cell source for in vitro investigation of hepatic drug metabolism and disposition. In this study, a multi-compartment capillary membrane-based bioreactor technology for three-dimensional (3D) perfusion culture was further developed and miniaturized to a volume of less than 0.5 ml to reduce demand for cells. The miniaturized bioreactor was composed of two capillary layers, each made of alternately arranged oxygen and medium capillaries serving as a 3D culture for the cells. Metabolic activity and stability of primary human hepatocytes was studied in this bioreactor in the presence of 2.5% fetal calf serum (FCS) under serum-free conditions over a culture period of 10 days. The miniaturized bioreactor showed functions comparable to previously reported data for larger variants. Glucose and lactate metabolism, urea production, albumin synthesis and release of intracellular enzymes (AST, ALT, GLDH) showed no significant differences between serum-free and serum-supplemented bioreactors. Activities of human-relevant cytochrome P450 (CYP) isoenzymes (CYP1A2, CYP3A4/5, CYP2C9, CYP2D6, CYP2B6) analyzed by determination of product formation rates from selective probe substrates were also comparable in both groups. Gene expression analysis showed moderately higher expression in the majority of CYP enzymes, transport proteins and enzymes of Phase II metabolism in the serum-free bioreactors compared to those maintained with FCS. In conclusion, the miniaturized bioreactor maintained stable function over the investigated period and thus provides a suitable system for pharmacological studies on primary human hepatocytes under defined serum-free conditions.

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

    Institute of Scientific and Technical Information of China (English)

    WEI Xin; LI Dao-bing; XU Feng; WANG Yan; ZHU Yu-chun; LI Hong; WANG Kun-jie

    2011-01-01

    Background 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.Methods 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.Results 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.Conclusions 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

  4. 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

  5. Establishing Liver Bioreactors for In Vitro Research.

    Science.gov (United States)

    Rebelo, Sofia P; Costa, Rita; Sousa, Marcos F Q; Brito, Catarina; Alves, Paula M

    2015-01-01

    In vitro systems that can effectively model liver function for long periods of time are fundamental tools for preclinical research. Nevertheless, the adoption of in vitro research tools at the earliest stages of drug development has been hampered by the lack of culture systems that offer the robustness, scalability, and flexibility necessary to meet industry's demands. Bioreactor-based technologies, such as stirred tank bioreactors, constitute a feasible approach to aggregate hepatic cells and maintain long-term three-dimensional cultures. These three-dimensional cultures sustain the polarity, differentiated phenotype, and metabolic performance of human hepatocytes. Culture in computer-controlled stirred tank bioreactors allows the maintenance of physiological conditions, such as pH, dissolved oxygen, and temperature, with minimal fluctuations. Moreover, by operating in perfusion mode, gradients of soluble factors and metabolic by-products can be established, aiming at resembling the in vivo microenvironment. This chapter provides a protocol for the aggregation and culture of hepatocyte spheroids in stirred tank bioreactors by applying perfusion mode for the long-term culture of human hepatocytes. This in vitro culture system is compatible with feeding high-throughput screening platforms for the assessment of drug elimination pathways, being a useful tool for toxicology research and drug development in the preclinical phase.

  6. Space Bioreactor Science Workshop

    Science.gov (United States)

    Morrison, Dennis R. (Editor)

    1987-01-01

    The first space bioreactor has been designed for microprocessor control, no gaseous headspace, circulation and resupply of culture medium, and a slow mixing in very low shear regimes. Various ground based bioreactors are being used to test reactor vessel design, on-line sensors, effects of shear, nutrient supply, and waste removal from continuous culture of human cells attached to microcarriers. The small (500 ml) bioreactor is being constructed for flight experiments in the Shuttle middeck to verify systems operation under microgravity conditions and to measure the efficiencies of mass transport, gas transfer, oxygen consumption, and control of low shear stress on cells. Applications of microcarrier cultures, development of the first space bioreactor flight system, shear and mixing effects on cells, process control, and methods to monitor cell metabolism and nutrient requirements are among the topics covered.

  7. Hydraulic model of a gas-lift bioreactor with flocculating yeast

    Energy Technology Data Exchange (ETDEWEB)

    Roca, E. [Chemical Engineering Dept., Univ. of Santiago de Compostela (Spain); Ghommidh, C. [G.B.S.A. Microbiologie et Biochimie Industrielle. Univ. Montpellier 2, 34 Montpellier (France); Navarro, J.M. [G.B.S.A. Microbiologie et Biochimie Industrielle. Univ. Montpellier 2, 34 Montpellier (France); Lema, J.M. [Chemical Engineering Dept., Univ. of Santiago de Compostela (Spain)

    1995-04-01

    The hydraulic model of a gas lift bioreactor, during a continuous alcoholic fermentation by using a strongly flocculating yeast, is analysed. Sucrose at two different concentrations (50 and 100 g/l) was used as substrate and the dilution rate for all the experiments was 1 h{sup -1}. The biomass concentrations were between 85 and 110 g dry weight/l. A stimulus response technique was used to obtain the Residence Time Distribution curves, a pulse of a lactose solution being used as the tracer. Mixing time was determined by means of the response to a pulse of an acid tracer. These experiments were carried out by using an on-line data-acquisition system. The bioreactor behaviour is completely homogeneous, except for high substrate and biomass concentrations. A two parameters combined model is necessary, in this case, to fit the experimental data. Mixing times are very low, in the order of 10 seconds. (orig.)

  8. Mathematical modelling and optimization of hydrogen continuous production in a fixed bed bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Palazzi, E.; Perego, P.; Fabiano, B. [University of Genoa, Genova (Italy). Chemical and Process Engineering Department ' G.B. Bonino'

    2002-09-01

    The purpose of this paper is to investigate, both theoretically and experimentally, hydrogen production from agro-industrial by-products using a continuous bioreactor packed with a mixture of spongy and glass beads and inoculated with Enterobacter aerogenes. Replicated series of experimental runs were performed to study the effects of residence time on hydrogen evolution rate and to characterize the critical conditions for the wash out, as a function of the inlet glucose concentration and of the fluid superficial velocity. A further series of experimental runs was focused on the effects of both residence time and inlet glucose concentration over hydrogen productivity. A kinetic model of the process was developed and showed good agreement with experimental data, thus representing a potential tool to design a large-scale fermenter. In fact, the model was applied to the optimal design of a bioreactor suitable of feeding a phosphoric acid fuel cell of a target power. (author)

  9. Modeling of submerged membrane bioreactor treating cheese whey wastewater by artificial neural network.

    Science.gov (United States)

    Cinar, Ozer; Hasar, Halil; Kinaci, Cumali

    2006-05-17

    A submerged membrane bioreactor receiving cheese whey was modeled by artificial neural network and its performance over a period of 100 days at different solids retention times was evaluated with this robust tool. A cascade-forward network was used to model the membrane bioreactor and normalization was used as a preprocessing method. The network was fed with two subsets of operational data, with two-thirds being used for training and one-third for testing the performance of the artificial neural network. The training procedure for effluent chemical oxygen demand (COD), ammonia, nitrate and total phosphate concentrations was very successful and a perfect match was obtained between the measured and the calculated concentrations. The results of the confirmation (or testing) procedure for effluent ammonia and nitrate concentrations were very successful; however, the results of the confirmation procedure for effluent COD and total phosphate concentrations were only satisfactory.

  10. Distributed and Lumped Parameter Models for the Characterization of High Throughput Bioreactors

    Science.gov (United States)

    Conoscenti, Gioacchino; Cutrì, Elena; Tuan, Rocky S.; Raimondi, Manuela T.; Gottardi, Riccardo

    2016-01-01

    Next generation bioreactors are being developed to generate multiple human cell-based tissue analogs within the same fluidic system, to better recapitulate the complexity and interconnection of human physiology [1, 2]. The effective development of these devices requires a solid understanding of their interconnected fluidics, to predict the transport of nutrients and waste through the constructs and improve the design accordingly. In this work, we focus on a specific model of bioreactor, with multiple input/outputs, aimed at generating osteochondral constructs, i.e., a biphasic construct in which one side is cartilaginous in nature, while the other is osseous. We next develop a general computational approach to model the microfluidics of a multi-chamber, interconnected system that may be applied to human-on-chip devices. This objective requires overcoming several challenges at the level of computational modeling. The main one consists of addressing the multi-physics nature of the problem that combines free flow in channels with hindered flow in porous media. Fluid dynamics is also coupled with advection-diffusion-reaction equations that model the transport of biomolecules throughout the system and their interaction with living tissues and C constructs. Ultimately, we aim at providing a predictive approach useful for the general organ-on-chip community. To this end, we have developed a lumped parameter approach that allows us to analyze the behavior of multi-unit bioreactor systems with modest computational effort, provided that the behavior of a single unit can be fully characterized. PMID:27669413

  11. Modeling the microbial growth and temperature profile in a fixed-bed bioreactor.

    Science.gov (United States)

    da Silveira, Christian L; Mazutti, Marcio A; Salau, Nina P G

    2014-10-01

    Aiming to scale up and apply control and optimization strategies, currently is required the development of accurate plant models to forecast the process nonlinear dynamics. In this work, a mathematical model to predict the growth of the Kluyveromyces marxianus and temperature profile in a fixed-bed bioreactor for solid-state fermentation using sugarcane bagasse as substrate was built up. A parameter estimation technique was performed to fit the mathematical model to the experimental data. The estimated parameters and the model fitness were evaluated with statistical analyses. The results have shown the estimated parameters significance, with 95 % confidence intervals, and the good quality of process model to reproduce the experimental data.

  12. Hybridoma cell-culture and glycan profile dataset at various bioreactor conditions.

    Science.gov (United States)

    Bhatia, Hemlata; Read, Erik; Agarabi, Cyrus; Brorson, Kurt; Lute, Scott; Yoon, Seongkyu

    2016-12-01

    This is an "11 factor-2 level-12 run" Plackett-Burman experimental design dataset. The dataset includes 11 engineering bioreactor parameters as input variables. These 11 factors were varied at 2 levels and 23 response variables that are glycan profile attributes, were measured "A Design Space Exploration for Control of Critical Quality Attributes of mAb" (H. Bhatia, E.K. Read, C.D. Agarabi, K.A. Brorson, S.C. Lute, S. Yoon S, 2016) [2].

  13. Comparison of the Modeling Approach between Membrane Bioreactor and Conventional Activated Sludge Processes

    DEFF Research Database (Denmark)

    Jiang, Tao; Sin, Gürkan; Spanjers, Henri

    2009-01-01

    Activated sludge models (ASM) have been developed and largely applied in conventional activated sludge (CAS) systems. The applicability of ASM to model membrane bioreactors (MBR) and the differences in modeling approaches have not been studied in detail. A laboratory-scale MBR was modeled using ASM...... to the inhibition effect of soluble microbial products (SMP) at elevated concentration. Second, a greater biomass affinity to oxygen and ammonium was found, which was probably related to smaller MBR sludge flocs. Finally, the membrane throughput during membrane backwashing/relaxation can be normalized...

  14. In vitro antifungal activity of extracts obtained from Hypericum perforatum adventitious roots cultured in a mist bioreactor against planktonic cells and biofilm of Malassezia furfur.

    Science.gov (United States)

    Simonetti, Giovanna; Tocci, Noemi; Valletta, Alessio; Brasili, Elisa; D'Auria, Felicia Diodata; Idoux, Alicia; Pasqua, Gabriella

    2016-01-01

    Xanthone-rich extracts from Hypericum perforatum root cultures grown in a Mist Bioreactor as antifungal agents against Malassezia furfur. Extracts of Hypericum perforatum roots grown in a bioreactor showed activity against planktonic cells and biofilm of Malassezia furfur. Dried biomass, obtained from roots grown under controlled conditions in a ROOTec mist bioreactor, has been extracted with solvents of increasing polarity (i.e. chloroform, ethyl acetate and methanol). The methanolic fraction was the richest in xanthones (2.86 ± 0.43 mg g(-1) DW) as revealed by HPLC. The minimal inhibitory concentration of the methanol extract against M. furfur planktonic cells was 16 μg mL(-1). The inhibition percentage of biofilm formation, at a concentration of 16 μg mL(-1), ranged from 14% to 39%. The results show that H. perforatum root extracts could be used as new antifungal agents in the treatment of Malassezia infections.

  15. Open source software to control Bioflo bioreactors.

    Directory of Open Access Journals (Sweden)

    David A Burdge

    Full Text Available Bioreactors are designed to support highly controlled environments for growth of tissues, cell cultures or microbial cultures. A variety of bioreactors are commercially available, often including sophisticated software to enhance the functionality of the bioreactor. However, experiments that the bioreactor hardware can support, but that were not envisioned during the software design cannot be performed without developing custom software. In addition, support for third party or custom designed auxiliary hardware is often sparse or absent. This work presents flexible open source freeware for the control of bioreactors of the Bioflo product family. The functionality of the software includes setpoint control, data logging, and protocol execution. Auxiliary hardware can be easily integrated and controlled through an integrated plugin interface without altering existing software. Simple experimental protocols can be entered as a CSV scripting file, and a Python-based protocol execution model is included for more demanding conditional experimental control. The software was designed to be a more flexible and free open source alternative to the commercially available solution. The source code and various auxiliary hardware plugins are publicly available for download from https://github.com/LibourelLab/BiofloSoftware. In addition to the source code, the software was compiled and packaged as a self-installing file for 32 and 64 bit windows operating systems. The compiled software will be able to control a Bioflo system, and will not require the installation of LabVIEW.

  16. Open source software to control Bioflo bioreactors.

    Science.gov (United States)

    Burdge, David A; Libourel, Igor G L

    2014-01-01

    Bioreactors are designed to support highly controlled environments for growth of tissues, cell cultures or microbial cultures. A variety of bioreactors are commercially available, often including sophisticated software to enhance the functionality of the bioreactor. However, experiments that the bioreactor hardware can support, but that were not envisioned during the software design cannot be performed without developing custom software. In addition, support for third party or custom designed auxiliary hardware is often sparse or absent. This work presents flexible open source freeware for the control of bioreactors of the Bioflo product family. The functionality of the software includes setpoint control, data logging, and protocol execution. Auxiliary hardware can be easily integrated and controlled through an integrated plugin interface without altering existing software. Simple experimental protocols can be entered as a CSV scripting file, and a Python-based protocol execution model is included for more demanding conditional experimental control. The software was designed to be a more flexible and free open source alternative to the commercially available solution. The source code and various auxiliary hardware plugins are publicly available for download from https://github.com/LibourelLab/BiofloSoftware. In addition to the source code, the software was compiled and packaged as a self-installing file for 32 and 64 bit windows operating systems. The compiled software will be able to control a Bioflo system, and will not require the installation of LabVIEW.

  17. A new dynamic model for highly efficient mass transfer in aerated bioreactors and consequences for kLa identification.

    Science.gov (United States)

    Müller, Stefan; Murray, Douglas B; Machne, Rainer

    2012-12-01

    Gas-liquid mass transfer is often rate-limiting in laboratory and industrial cultures of aerobic or autotrophic organisms. The volumetric mass transfer coefficient k(L) a is a crucial characteristic for comparing, optimizing, and upscaling mass transfer efficiency of bioreactors. Reliable dynamic models and resulting methods for parameter identification are needed for quantitative modeling of microbial growth dynamics. We describe a laboratory-scale stirred tank reactor (STR) with a highly efficient aeration system (k(L) a ≈ 570 h(-1)). The reactor can sustain yeast culture with high cell density and high oxygen uptake rate, leading to a significant drop in gas concentration from inflow to outflow (by 21%). Standard models fail to predict the observed mass transfer dynamics and to identify k(L) a correctly. In order to capture the concentration gradient in the gas phase, we refine a standard ordinary differential equation (ODE) model and obtain a system of partial integro-differential equations (PIDE), for which we derive an approximate analytical solution. Specific reactor configurations, in particular a relatively short bubble residence time, allow a quasi steady-state approximation of the PIDE system by a simpler ODE model which still accounts for the concentration gradient. Moreover, we perform an appropriate scaling of all variables and parameters. In particular, we introduce the dimensionless overall efficiency κ, which is more informative than k(L) a since it combines the effects of gas inflow, exchange, and solution. Current standard models of mass transfer in laboratory-scale aerated STRs neglect the gradient in the gas concentration, which arises from highly efficient bubbling systems and high cellular exchange rates. The resulting error in the identification of κ (and hence k(L) a) increases dramatically with increasing mass transfer efficiency. Notably, the error differs between cell-free and culture-based methods of parameter identification

  18. Screening of phenylpyruvic acid producers and optimization of culture conditions in bench scale bioreactors.

    Science.gov (United States)

    Coban, Hasan B; Demirci, Ali; Patterson, Paul H; Elias, Ryan J

    2014-11-01

    Alpha keto acids are deaminated forms of amino acids that have received significant attention as feed and food additives in the agriculture and medical industries. To date, their production has been commonly performed at shake-flask scale with low product concentrations. In this study, production of phenylpyruvic acid (PPA), which is the alpha keto acid of phenylalanine was investigated. First, various microorganisms were screened to select the most efficient producer. Thereafter, growth parameters (temperature, pH, and aeration) were optimized in bench scale bioreactors to maximize both PPA and biomass concentration in bench scale bioreactors, using response surface methodology. Among the four different microorganisms evaluated, Proteus vulgaris was the most productive strain for PPA production. Optimum temperature, pH, and aeration conditions were determined as 34.5 °C, 5.12, and 0.5 vvm for PPA production, whereas 36.9 °C, pH 6.87, and 0.96 vvm for the biomass production. Under these optimum conditions, PPA concentration was enhanced to 1,054 mg/L, which was almost three times higher than shake-flask fermentation concentrations. Moreover, P. vulgaris biomass was produced at 3.25 g/L under optimum conditions. Overall, this study demonstrated that optimization of growth parameters improved PPA production in 1-L working volume bench-scale bioreactors compared to previous studies in the literature and was a first step to scale up the production to industrial production.

  19. Hybridoma cell-culture and glycan profile dataset at various bioreactor conditions

    Directory of Open Access Journals (Sweden)

    Hemlata Bhatia

    2016-12-01

    Full Text Available This is an “11 factor-2 level-12 run” Plackett-Burman experimental design dataset. The dataset includes 11 engineering bioreactor parameters as input variables. These 11 factors were varied at 2 levels and 23 response variables that are glycan profile attributes, were measured “A Design Space Exploration for Control of Critical Quality Attributes of mAb” (H. Bhatia, E.K. Read, C.D. Agarabi, K.A. Brorson, S.C. Lute, S. Yoon S, 2016 [2].

  20. In silico multi-scale model of transport and dynamic seeding in a bone tissue engineering perfusion bioreactor.

    Science.gov (United States)

    Spencer, T J; Hidalgo-Bastida, L A; Cartmell, S H; Halliday, I; Care, C M

    2013-04-01

    Computer simulations can potentially be used to design, predict, and inform properties for tissue engineering perfusion bioreactors. In this work, we investigate the flow properties that result from a particular poly-L-lactide porous scaffold and a particular choice of perfusion bioreactor vessel design used in bone tissue engineering. We also propose a model to investigate the dynamic seeding properties such as the homogeneity (or lack of) of the cellular distribution within the scaffold of the perfusion bioreactor: a pre-requisite for the subsequent successful uniform growth of a viable bone tissue engineered construct. Flows inside geometrically complex scaffolds have been investigated previously and results shown at these pore scales. Here, it is our aim to show accurately that through the use of modern high performance computers that the bioreactor device scale that encloses a scaffold can affect the flows and stresses within the pores throughout the scaffold which has implications for bioreactor design, control, and use. Central to this work is that the boundary conditions are derived from micro computed tomography scans of both a device chamber and scaffold in order to avoid generalizations and uncertainties. Dynamic seeding methods have also been shown to provide certain advantages over static seeding methods. We propose here a novel coupled model for dynamic seeding accounting for flow, species mass transport and cell advection-diffusion-attachment tuned for bone tissue engineering. The model highlights the timescale differences between different species suggesting that traditional homogeneous porous flow models of transport must be applied with caution to perfusion bioreactors. Our in silico data illustrate the extent to which these experiments have the potential to contribute to future design and development of large-scale bioreactors.

  1. 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)

  2. Modeling Cultural Dynamics

    CERN Document Server

    Gabora, Liane

    2008-01-01

    EVOC (for EVOlution of Culture) is a computer model of culture that enables us to investigate how various factors such as barriers to cultural diffusion, the presence and choice of leaders, or changes in the ratio of innovation to imitation affect the diversity and effectiveness of ideas. It consists of neural network based agents that invent ideas for actions, and imitate neighbors' actions. The model is based on a theory of culture according to which what evolves through culture is not memes or artifacts, but the internal models of the world that give rise to them, and they evolve not through a Darwinian process of competitive exclusion but a Lamarckian process involving exchange of innovation protocols. EVOC shows an increase in mean fitness of actions over time, and an increase and then decrease in the diversity of actions. Diversity of actions is positively correlated with population size and density, and with barriers between populations. Slowly eroding borders increase fitness without sacrificing diver...

  3. Integration of Bioreactor and Membrane Separation Processes: A Model Based Approach

    DEFF Research Database (Denmark)

    Prado Rubio, Oscar Andres

    to as Reverse Electro-Enhanced Dialysis (REED). Unconsumed substrate and biomass are effectively recycled to the fermenter. In the second membrane separation stage, the lactate is recovered and concentrated as lactic acid using Electrodialysis with bipolar membranes (EDBM), while sodium hydroxide is regenerated...... test. Satisfactory results are obtained regulating the pH and managing the input constraints. The design and operability of the integrated bioreactor and REED module are investigated using the developed models and control structure. The study involves two different case studies: continuous lactic acid...

  4. Integration of Bioreactor and Membrane Separation Processes: A Model Based Approach

    DEFF Research Database (Denmark)

    Prado Rubio, Oscar Andres

    to as Reverse Electro-Enhanced Dialysis (REED). Unconsumed substrate and biomass are effectively recycled to the fermenter. In the second membrane separation stage, the lactate is recovered and concentrated as lactic acid using Electrodialysis with bipolar membranes (EDBM), while sodium hydroxide is regenerated...... test. Satisfactory results are obtained regulating the pH and managing the input constraints. The design and operability of the integrated bioreactor and REED module are investigated using the developed models and control structure. The study involves two different case studies: continuous lactic acid...

  5. Comparing and Contrasting Traditional Membrane Bioreactor Models with Novel Ones Based on Time Series Analysis

    Directory of Open Access Journals (Sweden)

    Parneet Paul

    2013-02-01

    Full Text Available The computer modelling and simulation of wastewater treatment plant and their specific technologies, such as membrane bioreactors (MBRs, are becoming increasingly useful to consultant engineers when designing, upgrading, retrofitting, operating and controlling these plant. This research uses traditional phenomenological mechanistic models based on MBR filtration and biochemical processes to measure the effectiveness of alternative and novel time series models based upon input–output system identification methods. Both model types are calibrated and validated using similar plant layouts and data sets derived for this purpose. Results prove that although both approaches have their advantages, they also have specific disadvantages as well. In conclusion, the MBR plant designer and/or operator who wishes to use good quality, calibrated models to gain a better understanding of their process, should carefully consider which model type is selected based upon on what their initial modelling objectives are. Each situation usually proves unique.

  6. Development of Hybrid Models for a Vapor-Phase Fungi Bioreactor

    Directory of Open Access Journals (Sweden)

    Giorgia Spigno

    2015-01-01

    Full Text Available This study is aimed at the development of a model for an experimental vapour-phase fungi bioreactor, which could be derived in a simple way using the available measurements of a pilot-plant reactor, without the development of ad hoc experiments for the evaluation of fungi kinetics and the estimation of parameters related to biofilm characteristics. The proposed approach is based on hybrid models, obtained by the connection of the mass balance equation (used in traditional phenomenological models with a feedforward neural network (used in black-box modelling, and the proper use of statistical tools for the model assessment and system understanding. Two different hybrid models were developed and compared by proper performance indexes, and their capability to predict the biological complex phenomena was demonstrated and compared to that of a first-principle model.

  7. A three-dimensional computational fluid dynamics model of shear stress distribution during neotissue growth in a perfusion bioreactor.

    Science.gov (United States)

    Guyot, Y; Luyten, F P; Schrooten, J; Papantoniou, I; Geris, L

    2015-12-01

    Bone tissue engineering strategies use flow through perfusion bioreactors to apply mechanical stimuli to cells seeded on porous scaffolds. Cells grow on the scaffold surface but also by bridging the scaffold pores leading a fully filled scaffold following the scaffold's geometric characteristics. Current computational fluid dynamic approaches for tissue engineering bioreactor systems have been mostly carried out for empty scaffolds. The effect of 3D cell growth and extracellular matrix formation (termed in this study as neotissue growth), on its surrounding fluid flow field is a challenge yet to be tackled. In this work a combined approach was followed linking curvature driven cell growth to fluid dynamics modeling. The level-set method (LSM) was employed to capture neotissue growth driven by curvature, while the Stokes and Darcy equations, combined in the Brinkman equation, provided information regarding the distribution of the shear stress profile at the neotissue/medium interface and within the neotissue itself during growth. The neotissue was assumed to be micro-porous allowing flow through its structure while at the same time allowing the simulation of complete scaffold filling without numerical convergence issues. The results show a significant difference in the amplitude of shear stress for cells located within the micro-porous neo-tissue or at the neotissue/medium interface, demonstrating the importance of taking along the neotissue in the calculation of the mechanical stimulation of cells during culture.The presented computational framework is used on different scaffold pore geometries demonstrating its potential to be used a design as tool for scaffold architecture taking into account the growing neotissue. Biotechnol. Bioeng. 2015;112: 2591-2600. © 2015 Wiley Periodicals, Inc.

  8. Modeling organic nitrogen conversions in activated sludge bioreactors.

    Science.gov (United States)

    Makinia, Jacek; Pagilla, Krishna; Czerwionka, Krzysztof; Stensel, H David

    2011-01-01

    For biological nutrient removal (BNR) systems designed to maximize nitrogen removal, the effluent total nitrogen (TN) concentration may range from 2.0 to 4.0 g N/m(3) with about 25-50% in the form of organic nitrogen (ON). In this study, current approaches to modeling organic N conversions (separate processes vs. constant contents of organic fractions) were compared. A new conceptual model of ON conversions was developed and combined with Activated Sludge Model No. 2d (ASM2d). The model addresses a new insight into the processes of ammonification, biomass decay and hydrolysis of particulate and colloidal ON (PON and CON, respectively). Three major ON fractions incorporated are defined as dissolved (DON) (model parameter set, the behaviors of both inorganic N forms (NH4-N, NOX-N) and ON forms (DON, CON) in the batch experiments were predicted. The challenges to accurately simulate and predict effluent ON levels from BNR systems are due to analytical methods of direct ON measurement (replacing TKN) and lack of large enough database (in-process measurements, dynamic variations of the ON concentrations) which can be used to determine parameter value ranges.

  9. 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.

  10. 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.

  11. A novel approach to recycle bacterial culture waste for fermentation reuse via a microbial fuel cell-membrane bioreactor system.

    Science.gov (United States)

    Li, Jian; Zhu, Yuan; Zhuang, Liangpeng; Otsuka, Yuichiro; Nakamura, Masaya; Goodell, Barry; Sonoki, Tomonori; He, Zhen

    2015-09-01

    Biochemical production processes require water and nutrient resources for culture media preparation, but aqueous waste is generated after the target products are extracted. In this study, culture waste (including cells) produced from a lab-scale fermenter was fed into a microbial fuel cell-membrane bioreactor (MFC-MBR) system. Electrical energy was generated via the interaction between the microbial consortia and the solid electrode in the MFC. The treated wastewater was reclaimed in this process which was reused as a solvent and a nutrient source in subsequent fermentation. Polarization testing showed that the MFC produced a maximum current density of 37.53 A m(-3) with a maximum power density of 5.49 W m(-3). The MFC was able to generate 0.04 kWh of energy per cubic meter of culture waste treated. The lab-scale fermenters containing pure cultures of an engineered Pseudomonas spp. were used to generate 2-pyrone-4,6-dicarboxylic acid (PDC), a high value platform chemical. When the MFC-MBR-treated wastewater was used for the fermenter culture medium, a specific bacterial growth rate of 1.00 ± 0.05 h(-1) was obtained with a PDC production rate of 708.11 ± 64.70 mg PDC L(-1) h(-1). Comparable values for controls using pure water were 0.95 ± 0.06 h(-1) and 621.01 ± 22.09 mg PDC L(-1) h(-1) (P > 0.05), respectively. The results provide insight on a new approach for more sustainable bio-material production while at the same time generating energy, and suggest that the treated wastewater can be used as a solvent and a nutrient source for the fermentation production of high value platform chemicals.

  12. a Cultural Market Model

    Science.gov (United States)

    HerdaǦDELEN, Amaç; Bingol, Haluk

    Social interactions and personal tastes shape our consumption behavior of cultural products. In this study, we present a computational model of a cultural market and we aim to analyze the behavior of the consumer population as an emergent phenomena. Our results suggest that the final market shares of cultural products dramatically depend on consumer heterogeneity and social interaction pressure. Furthermore, the relation between the resulting market shares and social interaction is robust with respect to a wide range of variation in the parameter values and the type of topology.

  13. A Cultural Market Model

    CERN Document Server

    Herdagdelen, Amac

    2008-01-01

    Social interactions and personal tastes shape our consumption behavior of cultural products. In this study, we present a computational model of a cultural market and we aim to analyze the behavior of the consumer population as an emergent phenomena. Our results suggest that the final market shares of cultural products dramatically depend on consumer heterogeneity and social interaction pressure. Furthermore, the relation between the resulting market shares and social interaction is robust with respect to a wide range of variation in the parameter values and the type of topology.

  14. Analysis of Gene Expression Signatures for Osteogenic 3D Perfusion-Bioreactor Cell Cultures Based on a Multifactorial DoE Approach

    Directory of Open Access Journals (Sweden)

    Ioannis Papantoniou

    2014-08-01

    Full Text Available The use of multifactorial design of experiments (DoE in tissue engineering bioprocess development will contribute to the robust manufacturing of tissue engineered constructs by linking their quality characteristics to bioprocess operating parameters. In this work, perfusion bioreactors were used for the in vitro culture and osteogenic differentiation of human periosteum-derived cells (hPDCs seeded on three-dimensional titanium (Ti alloy scaffolds. A CaP-supplemented medium was used to induce differentiation of the cultured hPDCs. A two-level, three-factor fractional factorial design was employed to evaluate a range of bioreactor operating conditions by changing the levels of the following parameters: flow rate (0.5–2 mL/min, cell culture duration (7–21 days and cell seeding density (1.5 × 103–3 × 103 cells/cm2. This approach allowed for evaluating the individual impact of the aforementioned process parameters upon a range of genes that are related to the osteogenic lineage, such as collagen type I, alkaline phosphatase, osterix, osteopontin and osteocalcin. Furthermore, by overlaying gene-specific response surfaces, an integrated operating process space was highlighted within which predetermined values of the six genes of interest (i.e., gene signature could be minimally met over the course of the bioreactor culture time.

  15. Kinetic model of continuous ethanol fermentation in closed-circulating process with pervaporation membrane bioreactor by Saccharomyces cerevisiae.

    Science.gov (United States)

    Fan, Senqing; Chen, Shiping; Tang, Xiaoyu; Xiao, Zeyi; Deng, Qing; Yao, Peina; Sun, Zhaopeng; Zhang, Yan; Chen, Chunyan

    2015-02-01

    Unstructured kinetic models were proposed to describe the principal kinetics involved in ethanol fermentation in a continuous and closed-circulating fermentation (CCCF) process with a pervaporation membrane bioreactor. After ethanol was removed in situ from the broth by the membrane pervaporation, the secondary metabolites accumulated in the broth became the inhibitors to cell growth. The cell death rate related to the deterioration of the culture environment was described as a function of the cell concentration and fermentation time. In CCCF process, 609.8 g L(-1) and 750.1 g L(-1) of ethanol production were obtained in the first run and second run, respectively. The modified Gompertz model, correlating the ethanol production with the fermentation period, could be used to describe the ethanol production during CCCF process. The fitting results by the models showed good agreement with the experimental data. These models could be employed for the CCCF process technology development for ethanol fermentation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. The Culture Based Model: Constructing a Model of Culture

    Science.gov (United States)

    Young, Patricia A.

    2008-01-01

    Recent trends reveal that models of culture aid in mapping the design and analysis of information and communication technologies. Therefore, models of culture are powerful tools to guide the building of instructional products and services. This research examines the construction of the culture based model (CBM), a model of culture that evolved…

  17. Development of a Comprehensive Fouling Model for a Rotating Membrane Bioreactor System Treating Wastewater

    Directory of Open Access Journals (Sweden)

    Parneet Paul

    2015-01-01

    Full Text Available Membrane bioreactors (MBRs are now main stream wastewater treatment technologies. In recent times, novel pressure driven rotating membrane disc modules have been specially developed that induce high shear on the membrane surface, thereby reducing fouling. Previous research has produced dead-end filtration fouling model which combines all three classical mechanisms that was later used by another researcher as a starting point for a greatly refined model of a cross flow side-stream MBR that incorporated both hydrodynamics and soluble microbial products’ (SMP effects. In this study, a comprehensive fouling model was created based on this earlier work that incorporated all three classical fouling mechanisms for a rotating MBR system. It was tested and validated for best fit using appropriate data sets. The initial model fit appeared good for all simulations, although it still needs to be calibrated using further appropriate data sets.

  18. A versatile modular bioreactor platform for Tissue Engineering.

    Science.gov (United States)

    Schuerlein, Sebastian; Schwarz, Thomas; Krziminski, Steffan; Gätzner, Sabine; Hoppensack, Anke; Schwedhelm, Ivo; Schweinlin, Matthias; Walles, Heike; Hansmann, Jan

    2017-02-01

    Tissue Engineering (TE) bears potential to overcome the persistent shortage of donor organs in transplantation medicine. Additionally, TE products are applied as human test systems in pharmaceutical research to close the gap between animal testing and the administration of drugs to human subjects in clinical trials. However, generating a tissue requires complex culture conditions provided by bioreactors. Currently, the translation of TE technologies into clinical and industrial applications is limited due to a wide range of different tissue-specific, non-disposable bioreactor systems. To ensure a high level of standardization, a suitable cost-effectiveness, and a safe graft production, a generic modular bioreactor platform was developed. Functional modules provide robust control of culture processes, e.g. medium transport, gas exchange, heating, or trapping of floating air bubbles. Characterization revealed improved performance of the modules in comparison to traditional cell culture equipment such as incubators, or peristaltic pumps. By combining the modules, a broad range of culture conditions can be achieved. The novel bioreactor platform allows using disposable components and facilitates tissue culture in closed fluidic systems. By sustaining native carotid arteries, engineering a blood vessel, and generating intestinal tissue models according to a previously published protocol the feasibility and performance of the bioreactor platform was demonstrated. © 2017 The Authors. Biotechnology Journal published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Enhanced biological phosphorus removal and its modeling for the activated sludge and membrane bioreactor processes.

    Science.gov (United States)

    Zuthi, M F R; Guo, W S; Ngo, H H; Nghiem, L D; Hai, F I

    2013-07-01

    A modified activated sludge process (ASP) for enhanced biological phosphorus removal (EBPR) needs to sustain stable performance for wastewater treatment to avoid eutrophication in the aquatic environment. Unfortunately, the overall efficiency of the EBPR in ASPs and membrane bioreactors (MBRs) is frequently hindered by different operational/system constraints. Moreover, although phosphorus removal data from several wastewater treatment systems are available, a comprehensive mathematical model of the process is still lacking. This paper presents a critical review that highlights the core issues of the biological phosphorus removal in ASPs and MBRs while discussing the inhibitory process requirements for other nutrients' removal. This mini review also successfully provided an assessment of the available models for predicting phosphorus removal in both ASP and MBR systems. The advantages and limitations of the existing models were discussed together with the inclusion of few guidelines for their improvement. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. 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.

  1. Modelling, Optimization and Optimal Control of Small Scale Stirred Tank Bioreactors

    Directory of Open Access Journals (Sweden)

    Mitko Petrov

    2004-10-01

    Full Text Available Models of the mass-transfer in a stirred tank bioreactor depending on general indexes of the processes of aeration and mixing in concrete simplifications of the hydrodynamic structure of the flows are developed. The offered combined model after parameters identification is used for optimization of the parameters of the apparatus construction. The optimization problem is solved by using of the fuzzy sets theory and in this way the unspecified as a result of the model simplification are read. In conclusion an optimal control of a fed-batch fermentation process of E. coli is completed by using Neuro-Dynamic programming. The received results after optimization show a considerable improvement of the mass-transfer indexes and the quantity indexes at the end of the process.

  2. Stirred tank bioreactor culture combined with serum-/xenogeneic-free culture medium enables an efficient expansion of umbilical cord-derived mesenchymal stem/stromal cells.

    Science.gov (United States)

    Mizukami, Amanda; Fernandes-Platzgummer, Ana; Carmelo, Joana G; Swiech, Kamilla; Covas, Dimas T; Cabral, Joaquim M S; da Silva, Cláudia L

    2016-08-01

    Mesenchymal stem/stromal cells (MSC) are being widely explored as promising candidates for cell-based therapies. Among the different human MSC origins exploited, umbilical cord represents an attractive and readily available source of MSC that involves a non-invasive collection procedure. In order to achieve relevant cell numbers of human MSC for clinical applications, it is crucial to develop scalable culture systems that allow bioprocess control and monitoring, combined with the use of serum/xenogeneic (xeno)-free culture media. In the present study, we firstly established a spinner flask culture system combining gelatin-based Cultispher(®) S microcarriers and xeno-free culture medium for the expansion of umbilical cord matrix (UCM)-derived MSC. This system enabled the production of 2.4 (±1.1) x10(5) cells/mL (n = 4) after 5 days of culture, corresponding to a 5.3 (±1.6)-fold increase in cell number. The established protocol was then implemented in a stirred-tank bioreactor (800 mL working volume) (n = 3) yielding 115 million cells after 4 days. Upon expansion under stirred conditions, cells retained their differentiation ability and immunomodulatory potential. The development of a scalable microcarrier-based stirred culture system, using xeno-free culture medium that suits the intrinsic features of UCM-derived MSC represents an important step towards a GMP compliant large-scale production platform for these promising cell therapy candidates. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Production of Limonoids with Insect Antifeedant Activity in a Two-Stage Bioreactor Process with Cell Suspension Culture of Azadirachta indica.

    Science.gov (United States)

    Vásquez-Rivera, Andrés; Chicaiza-Finley, Diego; Hoyos, Rodrigo A; Orozco-Sánchez, Fernando

    2015-09-01

    Neem tree (Azadirachta indica) cell suspension culture is an alternative for the production of limonoids for insect control that overcomes limitations related to the supply of neem seeds. To establish conditions for cell growth and azadiracthin-related limonoid production, the effect of different sucrose concentrations, nitrate and phosphate in Murashige and Skoog (MS) medium, and the addition of one precursor and three elicitors was evaluated in shake flasks. The process was scaled up to a 3-l stirred tank bioreactor in one- and two-stage batch cultivation. In shake flasks, more than fivefold increase in the production of limonoids with the modified MS medium was observed (increase from 0.77 to 4.52 mg limonoids/g dry cell weight, DCW), while an increase of more than fourfold was achieved by adding the elicitors chitosan, salicylic acid, and jasmonic acid together (increase from 1.03 to 4.32 mg limonoids/g DCW). In the bioreactor, the volumetric production of limonoids was increased more than threefold with a two-stage culture in day 18 (13.82 mg limonoids/l in control single-stage process and 41.44 mg/l in two-stage process). The cultivation and operating mode of the bioreactor reported in this study may be adapted and used in optimization and process plant development for production of insect antifeedant limonoids with A. indica cell suspension cultures.

  4. Development of a compartment model based on CFD simulations for description of mixing in bioreactors

    Directory of Open Access Journals (Sweden)

    Crine, M.

    2010-01-01

    Full Text Available Understanding and modeling the complex interactions between biological reaction and hydrodynamics are a key problem when dealing with bioprocesses. It is fundamental to be able to accurately predict the hydrodynamics behavior of bioreactors of different size and its interaction with the biological reaction. CFD can provide detailed modeling about hydrodynamics and mixing. However, it is computationally intensive, especially when reactions are taken into account. Another way to predict hydrodynamics is the use of "Compartment" or "Multi-zone" models which are much less demanding in computation time than CFD. However, compartments and fluxes between them are often defined by considering global quantities not representative of the flow. To overcome the limitations of these two methods, a solution is to combine compartment modeling and CFD simulations. Therefore, the aim of this study is to develop a methodology in order to propose a compartment model based on CFD simulations of a bioreactor. The flow rate between two compartments can be easily computed from the velocity fields obtained by CFD. The difficulty lies in the definition of the zones in such a way they can be considered as perfectly mixed. The creation of the model compartments from CFD cells can be achieved manually or automatically. The manual zoning consists in aggregating CFD cells according to the user's wish. The automatic zoning defines compartments as regions within which the value of one or several properties are uniform with respect to a given tolerance. Both manual and automatic zoning methods have been developed and compared by simulating the mixing of an inert scalar. For the automatic zoning, several algorithms and different flow properties have been tested as criteria for the compartment creation.

  5. Characterization of TAP Ambr 250 disposable bioreactors, as a reliable scale-down model for biologics process development.

    Science.gov (United States)

    Xu, Ping; Clark, Colleen; Ryder, Todd; Sparks, Colleen; Zhou, Jiping; Wang, Michelle; Russell, Reb; Scott, Charo

    2017-03-01

    Demands for development of biological therapies is rapidly increasing, as is the drive to reduce time to patient. In order to speed up development, the disposable Automated Microscale Bioreactor (Ambr 250) system is increasingly gaining interest due to its advantages, including highly automated control, high throughput capacity, and short turnaround time. Traditional early stage upstream process development conducted in 2 - 5 L bench-top bioreactors requires high foot-print, and running cost. The establishment of the Ambr 250 as a scale-down model leads to many benefits in process development. In this study, a comprehensive characterization of mass transfer coefficient (kL a) in the Ambr 250 was conducted to define optimal operational conditions. Scale-down approaches, including dimensionless volumetric flow rate (vvm), power per unit volume (P/V) and kL a have been evaluated using different cell lines. This study demonstrates that the Ambr 250 generated comparable profiles of cell growth and protein production, as seen at 5-L and 1000-L bioreactor scales, when using kL a as a scale-down parameter. In addition to mimicking processes at large scales, the suitability of the Ambr 250 as a tool for clone selection, which is traditionally conducted in bench-top bioreactors, was investigated. Data show that cell growth, productivity, metabolite profiles, and product qualities of material generated using the Ambr 250 were comparable to those from 5-L bioreactors. Therefore, Ambr 250 can be used for clone selection and process development as a replacement for traditional bench-top bioreactors minimizing resource utilization during the early stages of development in the biopharmaceutical industry. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:478-489, 2017. © 2017 American Institute of Chemical Engineers.

  6. Progress in bioreactors of bioartiifcial livers

    Institute of Scientific and Technical Information of China (English)

    Cheng-Bo Yu; Xiao-Ping Pan; Lan-Juan Li

    2009-01-01

    BACKGROUND: Bioartiifcial liver support systems are becoming an effective therapy for hepatic failure. Bioreactors, as key devices in these systems, can provide a favorable growth and metabolic environment, mass exchange, and immunological isolation as a platform. Currently, stagnancy in bioreactor research is the main factor restricting the development of bioartiifcial liver support systems. DATA SOURCES: A PubMed database search of English-language literature was performed to identify relevant articles using the keywords "bioreactor", "bioartiifcial liver", "hepatocyte", and "liver failure". More than 40 articles related to the bioreactors of bioartiifcial livers were reviewed. RESULTS: Some progress has been made in the improvement of structures, functions, and modiifed macromolecular materials related to bioreactors in recent years. The current data on the improvement of bioreactor conifgurations for bioartiifcial livers or on the potential of the use of certain scaffold materials in bioreactors, combined with the clinical efifcacy and safety evaluation of cultured hepatocytesin vitro, indicate that the AMC (Academic Medical Center) BAL bioreactor and MELS (modular extracorporeal liver support) BAL bioreactor system can partly replace the synthetic and metabolic functions of the liver in phaseⅠ clinical studies. In addition, it has been indicated that the microlfuidic PDMS (polydimethylsiloxane) bioreactor, or SlideBioreactor, and the microfabricated grooved bioreactor are appropriate for hepatocyte culture, which is also promising for bioartiifcial livers. Similarly, modiifed scaffolds can promote the adhesion, growth, and function of hepatocytes, and provide reliable materials for bioreactors.CONCLUSIONS: Bioreactors, as key devices in bioartiifcial livers, play an important role in the therapy for liver failure both now and in the future. Bioreactor conifgurations are indispensable for the development of bioartiifcial livers used for liver

  7. Production of high-titer human influenza A virus with adherent and suspension MDCK cells cultured in a single-use hollow fiber bioreactor.

    Science.gov (United States)

    Tapia, Felipe; Vogel, Thomas; Genzel, Yvonne; Behrendt, Ilona; Hirschel, Mark; Gangemi, J David; Reichl, Udo

    2014-02-12

    Hollow fiber bioreactors (HFBRs) have been widely described as capable of supporting the production of highly concentrated monoclonal antibodies and recombinant proteins. Only recently HFBRs have been proposed as new single-use platforms for production of high-titer influenza A virus. These bioreactors contain multiple hollow fiber capillary tubes that separate the bioreactor in an intra- and an extra-capillary space. Cells are usually cultured in the extra-capillary space and can grow to a very high cell concentration. This work describes the evaluation of the single-use hollow fiber bioreactor PRIMER HF (Biovest International Inc., USA) for production of influenza A virus. The process was setup, characterized and optimized by running a total of 15 cultivations. The HFBRs were seeded with either adherent or suspension MDCK cells, and infected with influenza virus A/PR/8/34 (H1N1), and the pandemic strain A/Mexico/4108/2009 (H1N1). High HA titers and TCID₅₀ of up to 3.87 log₁₀(HA units/100 μL) and 1.8 × 10(10)virions/mL, respectively, were obtained for A/PR/8/34 influenza strain. Influenza virus was collected by performing multiple harvests of the extra-capillary space during a virus production time of up to 12 days. Cell-specific virus yields between 2,000 and 8,000 virions/cell were estimated for adherent MDCK cells, and between 11,000 and 19,000 virions/cell for suspension MDCK.SUS2 cells. These results do not only coincide with the cell-specific virus yields obtained with cultivations in stirred tank bioreactors and other high cell density systems, but also demonstrate that HFBRs are promising and competitive single-use platforms that can be considered for commercial production of influenza virus.

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

    Science.gov (United States)

    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

  9. Induction of secondary metabolism of Aspergillus terreus ATCC 20542 in the batch bioreactor cultures.

    Science.gov (United States)

    Boruta, Tomasz; Bizukojc, Marcin

    2016-04-01

    Cultivation of Aspergillus terreus ATCC 20542 in a stirred tank bioreactor was performed to induce the biosynthesis of secondary metabolites and provide the bioprocess-related insights into the metabolic capabilities of the investigated strain. The activation of biosynthetic routes was attempted by the diversification of process conditions and growth media. Several strategies were tested, including the addition of rapeseed oil or inulin, changing the concentration of nitrogen source, reduction of chlorine supply, cultivation under saline conditions, and using various aeration schemes. Fifteen secondary metabolites were identified in the course of the study by using ultra-high performance liquid chromatography coupled with mass spectrometry, namely mevinolinic acid, 4a,5-dihydromevinolinic acid, 3α-hydroxy-3,5-dihydromonacolin L acid, terrein, aspulvinone E, dihydroisoflavipucine, (+)-geodin, (+)-bisdechlorogeodin, (+)-erdin, asterric acid, butyrolactone I, desmethylsulochrin, questin, sulochrin, and demethylasterric acid. The study also presents the collection of mass spectra that can serve as a resource for future experiments. The growth in a salt-rich environment turned out to be strongly inhibitory for secondary metabolism and the formation of dense and compact pellets was observed. Generally, the addition of inulin, reducing the oxygen supply, and increasing the content of nitrogen source did not enhance the production of examined molecules. The most successful strategy involved the addition of rapeseed oil to the chlorine-deficient medium. Under these conditions, the highest levels of butyrolactone I, asterric acid, and mevinolinic acid were achieved and the presence of desmethylsulochrin and (+)-bisdechlorogeodin was detected in the broth. The constant and relatively high aeration rate in the idiophase was shown to be beneficial for terrein and (+)-geodin biosynthesis.

  10. Comparative study of withanolide production and the related transcriptional responses of biosynthetic genes in fungi elicited cell suspension culture of Withania somnifera in shake flask and bioreactor.

    Science.gov (United States)

    Ahlawat, Seema; Saxena, Parul; Ali, Athar; Khan, Shazia; Abdin, Malik Z

    2017-05-01

    Ashwagandha (Withania somnifera) is one of the most reputed medicinal plants in the traditional medicinal system. In this study, cell suspension culture of W. somnifera was elicited with cell homogenates of fungi (A. alternata, F. solani, V. dahliae and P. indica) in shake flask and the major withanolides like withanolide A, withaferin A and withanone were analysed. Simultaneously expression levels of key pathway genes from withanolides biosynthetic pathways were also checked via quantitative PCR in shake flask as well as in bioreactor. The results show that highest gene expression of 10.8, 5.8, 4.9, and 3.3 folds were observed with HMGR among all the expressed genes in cell suspension cultures with cell homogenates of 3% P. indica, 5% V. dahliae, 3% A. alternata and 3% F. solani, respectively, in comparison to the control in shake flask. Optimized concentration of cell homogenate of P. indica (3% v/v) was added to the growing culture in 5.0-l bioreactor under optimized up-scaling conditions and harvested after 22 days. The genes of MVA, MEP and withanolides biosynthetic pathways like HMGR, SS, SE, CAS, FPPS, DXR and DXS were up-regulated by 12.5, 4.9, 2.18, 4.65, 2.34, 1.89 and 1.4 folds, respectively in bioreactor. The enhancement of biomass (1.13 fold) and withanolides [withanolide A (1.7), withaferin A (1.5), and withanone (1.5) folds] in bioreactor in comparison to shake flask was also found to be in line with the up-regulation of genes of withanolide biosynthetic pathways. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  11. 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.

  12. 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.

  13. Morphological studies on the culture of kidney epithelial cells in a fiber-in-fiber bioreactor design with hollow fiber membranes.

    Science.gov (United States)

    Fey-Lamprecht, F; Albrecht, W; Groth, T; Weigel, T; Gross, U

    2003-05-01

    A hollow fiber-in-fiber-based bioreactor system was tested for the applicability to host kidney epithelial cells as a model system for a bioartificial kidney. Hollow fibers were prepared from polyacrylonitrile (PAN), polysulfone-polyvinylpyrollidinone (PVP) blend (PSU) and poly(acrylonitrile-N-vinylpyrollidinone) copolymer P(AN-NVP). Hollow fibers with smaller and larger diameters were prepared so that the smaller fitted into the larger, with a distance of 50-100 microm in between. The following material combinations as outer and inner fiber were applied: PAN-PAN; PSU-PSU, PSU-P(AN-NVP). Madin-Darby kidney epithelial cells (MDCK) were seeded in the interfiber space and cultured for a period up to 14 days. Light, scanning, and transmission electron microscopy were used to follow the adhesion and growth of cells, and to characterize their morphology. As a result, we found that MDCK cells were able to grow in the interfiber space in mono- and multilayers without signs of systemic degeneration. Comparison of the different materials showed that PAN and P(AN-NVP) provided the best growth conditions, indicated by a tight attachment of cells on hollow fiber membrane, and subsequent proliferation and development of structural elements of normal epithelia, such as tight junctions and microvilli. In conclusion, the fiber-in-fiber design seems to be an interesting system for the construction of a bioartificial kidney. Copyright 2003 Wiley Periodicals, Inc.

  14. Modeling and mitigation of denitrification 'woodchip' bioreactor phosphorus releases during treatment of aquaculture wastewater

    Science.gov (United States)

    Denitrification 'woodchip' bioreactors designed to remove nitrate from agricultural waters may either be phosphorus sources or sinks. A 24 d batch test showed woodchip leaching is an important source of phosphorus during bioreactor start-up with a leaching potential of approximately 20 -30 mg P per ...

  15. Fed-batch bioreactor process scale-up from 3-L to 2,500-L scale for monoclonal antibody production from cell culture.

    Science.gov (United States)

    Yang, Jeng-Dar; Lu, Canghai; Stasny, Brad; Henley, Joseph; Guinto, Woodrow; Gonzalez, Carlos; Gleason, Joseph; Fung, Monica; Collopy, Brett; Benjamino, Michael; Gangi, Jennifer; Hanson, Melissa; Ille, Elisabeth

    2007-09-01

    This case study focuses on the scale-up of a Sp2/0 mouse myeloma cell line based fed-batch bioreactor process, from the initial 3-L bench scale to the 2,500-L scale. A stepwise scale-up strategy that involved several intermediate steps in increasing the bioreactor volume was adopted to minimize the risks associated with scale-up processes. Careful selection of several available mixing models from literature, and appropriately applying the calculated results to our settings, resulted in successful scale-up of agitation speed for the large bioreactors. Consideration was also given to scale-up of the nutrient feeding, inoculation, and the set-points of operational parameters such as temperature, pH, dissolved oxygen, dissolved carbon dioxide, and aeration in an integrated manner. It has been demonstrated through the qualitative and the quantitative side-by-side comparison of bioreactor performance as well as through a panel of biochemical characterization tests that the comparability of the process and the product was well controlled and maintained during the process scale-up. The 2,500-L process is currently in use for the routine clinical production of Epratuzumab in support of two global Phase III clinical trials in patients with lupus. Today, the 2,500 L, fed-batch production process for Epratuzumab has met all scheduled batch releases, and the quality of the antibody is consistent and reproducible, meeting all specifications, thus confirming the robustness of the process.

  16. Delineating Cultural Models

    Science.gov (United States)

    2010-08-25

    cultural models of illness, mind, and body in stories concerning the temporomandibular joint (TMJ). Social Science and Medicine, 38(6), 775-788...includes a ba- sic description of the research effort, an in-depth review regarding research on opinion dynam- ics, a bibliography of collected research...et al., (2008) Salzarulo, (2006) Description Opinion is a binary state Opinion is one value from a finite discrete set Opinion is a set of

  17. Mathematical modeling of continuous ethanol fermentation in a membrane bioreactor by pervaporation compared to conventional system: Genetic algorithm.

    Science.gov (United States)

    Esfahanian, Mehri; Shokuhi Rad, Ali; Khoshhal, Saeed; Najafpour, Ghasem; Asghari, Behnam

    2016-07-01

    In this paper, genetic algorithm was used to investigate mathematical modeling of ethanol fermentation in a continuous conventional bioreactor (CCBR) and a continuous membrane bioreactor (CMBR) by ethanol permselective polydimethylsiloxane (PDMS) membrane. A lab scale CMBR with medium glucose concentration of 100gL(-1) and Saccharomyces cerevisiae microorganism was designed and fabricated. At dilution rate of 0.14h(-1), maximum specific cell growth rate and productivity of 0.27h(-1) and 6.49gL(-1)h(-1) were respectively found in CMBR. However, at very high dilution rate, the performance of CMBR was quite similar to conventional fermentation on account of insufficient incubation time. In both systems, genetic algorithm modeling of cell growth, ethanol production and glucose concentration were conducted based on Monod and Moser kinetic models during each retention time at unsteady condition. The results showed that Moser kinetic model was more satisfactory and desirable than Monod model.

  18. A plant-wide energy model for wastewater treatment plants: application to anaerobic membrane bioreactor technology.

    Science.gov (United States)

    Pretel, R; Robles, A; Ruano, M V; Seco, A; Ferrer, J

    2016-09-01

    The aim of this study is to propose a detailed and comprehensive plant-wide model for assessing the energy demand of different wastewater treatment systems (beyond the traditional activated sludge) in both steady- and unsteady-state conditions. The proposed model makes it possible to calculate power and heat requirements (W and Q, respectively), and to recover both power and heat from methane and hydrogen capture. In order to account for the effect of biological processes on heat requirements, the model has been coupled to the extended version of the BNRM2 plant-wide mathematical model, which is implemented in DESSAS simulation software. Two case studies have been evaluated to assess the model's performance: (1) modelling the energy demand of two urban wastewater treatment plants based on conventional activated sludge and submerged anaerobic membrane bioreactor (AnMBR) technologies in steady-state conditions and (2) modelling the dynamics of reactor temperature and heat requirements in an AnMBR plant in unsteady-state conditions. The results indicate that the proposed model can be used to assess the energy performance of different wastewater treatment processes and would thus be useful, for example, WWTP design or upgrading or the development of new control strategies for energy savings.

  19. Design challenges for space bioreactors

    Science.gov (United States)

    Seshan, P. K.; Petersen, G. R.

    1989-01-01

    The design of bioreactors for operation under conditions of microgravity presents problems and challenges. Absence of a significant body force such as gravity can have profound consequences for interfacial phenomena. Marangoni convection can no longer be overlooked. Many speculations on the advantages and benefits of microgravity can be found in the literature. Initial bioreactor research considerations for space applications had little regard for the suitability of the designs for conditions of microgravity. Bioreactors can be classified in terms of their function and type of operation. The complex interaction of parameters leading to optimal design and operation of a bioreactor is illustrated by the JSC mammalian cell culture system. The design of a bioreactor is strongly dependent upon its intended use as a production unit for cell mass and/or biologicals or as a research reactor for the study of cell growth and function. Therefore a variety of bioreactor configurations are presented in rapid summary. Following this, a rationale is presented for not attempting to derive key design parameters such as the oxygen transfer coefficient from ground-based data. A set of themes/objectives for flight experiments to develop the expertise for design of space bioreactors is then proposed for discussion. These experiments, carried out systematically, will provide a database from which engineering tools for space bioreactor design will be derived.

  20. The growth and saponin production of Platycodon grandiflorum (Jacq. A. DC. (Chinese bellflower hairy roots cultures maintained in shake flasks and mist bioreactor

    Directory of Open Access Journals (Sweden)

    Natalia Urbańska

    2014-07-01

    Full Text Available The growth and saponin accumulation were measured in two lines of transgenic hairy roots of Platycodon grandiflorum, Pl 6 and Pl 17, cultured for 8 weeks in 250-ml shake flasks containing 50 ml of hormone-free woody plant medium supplemented with 40 g/l sucrose and in the Pl 17 line cultured for 12 weeks in a 5-l mist bioreactor containing 1.5 l of the same medium. With both methods, the growth of transgenic hairy roots was assessed as both fresh and dry weight and the biomass growth was correlated with the conductivity and sucrose uptake. The accumulation of saponins was measured and compared with that in roots derived from the field cultivation. The saponin concentrations were significantly higher in the two hairy root lines cultured in shake flasks [6.92 g/100 g d.w. (g% and 5.82 g% in Pl 6 and Pl 17, respectively] and the line cultured in the bioreactor (5.93 g% than in the roots derived from the field cultivation (4.02 g%. The results suggest that cultures of P. grandiflorum hairy roots may be a valuable source for obtaining saponins.

  1. Use of Model-Based Nutrient Feeding for Improved Production of Artemisinin by Hairy Roots of Artemisia Annua in a Modified Stirred Tank Bioreactor.

    Science.gov (United States)

    Patra, Nivedita; Srivastava, Ashok K

    2015-09-01

    Artemisinin has been indicated to be a potent drug for the cure of malaria. Batch growth and artemisinin production kinetics of hairy root cultures of Artemisia annua were studied under shake flask conditions which resulted in accumulation of 12.49 g/L biomass and 0.27 mg/g artemisinin. Using the kinetic data, a mathematical model was identified to understand and optimize the system behavior. The developed model was then extrapolated to design nutrient feeding strategies during fed-batch cultivation for enhanced production of artemisinin. In one of the fed-batch cultivation, sucrose (37 g/L) feeding was done at a constant feed rate of 0.1 L/day during 10-15 days, which led to improved artemisinin accumulation of 0.77 mg/g. The second strategy of fed-batch hairy root cultivation involved maintenance of pseudo-steady state sucrose concentration (20.8 g/L) during 10-15 days which resulted in artemisinin accumulation of 0.99 mg/g. Fed-batch cultivation (with the maintenance of pseudo-steady state of substrate) of Artemisia annua hairy roots was, thereafter, implemented in bioreactor cultivation, which featured artemisinin accumulation of 1.0 mg/g artemisinin in 16 days of cultivation. This is the highest reported artemisinin yield by hairy root cultivation in a bioreactor.

  2. Validation of computational non-Newtonian fluid model for membrane bioreactor.

    Science.gov (United States)

    Sørensen, Lasse; Bentzen, Thomas Ruby; Skov, Kristian

    2015-01-01

    Membrane bioreactor (MBR) systems are often considered as the wastewater treatment method of the future due to their high effluent quality. One of the main problems with such systems is a relative large energy consumption, compared to conventional activated sludge (CAS) systems, which has led to further research in this specific area. A powerful tool for optimizing MBR-systems is computational fluid dynamics (CFD) modelling, which gives researchers the ability to describe the flow in the systems. A parameter which is often neglected in such models is the non-Newtonian properties of active sludge, which is of great importance for MBR systems since they operate at sludge concentrations up to a factor of 10 compared to CAS systems, resulting in strongly shear thinning liquids. A CFD-model is validated against measurements conducted in a system with rotating cross-flow membranes submerged in non-Newtonian liquids, where tangential velocities are measured with a Laser Doppler Anemometer (LDA). The CFD model is found to be capable of modelling the correct velocities in a range of setups, making CFD models a powerful tool for optimization of MBR systems.

  3. Factorial experimental design for the culture of human embryonic stem cells as aggregates in stirred suspension bioreactors reveals the potential for interaction effects between bioprocess parameters.

    Science.gov (United States)

    Hunt, Megan M; Meng, Guoliang; Rancourt, Derrick E; Gates, Ian D; Kallos, Michael S

    2014-01-01

    Traditional optimization of culture parameters for the large-scale culture of human embryonic stem cells (ESCs) as aggregates is carried out in a stepwise manner whereby the effect of varying each culture parameter is investigated individually. However, as evidenced by the wide range of published protocols and culture performance indicators (growth rates, pluripotency marker expression, etc.), there is a lack of systematic investigation into the true effect of varying culture parameters especially with respect to potential interactions between culture variables. Here we describe the design and execution of a two-parameter, three-level (3(2)) factorial experiment resulting in nine conditions that were run in duplicate 125-mL stirred suspension bioreactors. The two parameters investigated here were inoculation density and agitation rate, which are easily controlled, but currently, poorly characterized. Cell readouts analyzed included fold expansion, maximum density, and exponential growth rate. Our results reveal that the choice of best case culture parameters was dependent on which cell property was chosen as the primary output variable. Subsequent statistical analyses via two-way analysis of variance indicated significant interaction effects between inoculation density and agitation rate specifically in the case of exponential growth rates. Results indicate that stepwise optimization has the potential to miss out on the true optimal case. In addition, choosing an optimum condition for a culture output of interest from the factorial design yielded similar results when repeated with the same cell line indicating reproducibility. We finally validated that human ESCs remain pluripotent in suspension culture as aggregates under our optimal conditions and maintain their differentiation capabilities as well as a stable karyotype and strong expression levels of specific human ESC markers over several passages in suspension bioreactors.

  4. A study of murine bone marrow cells cultured in bioreactors which create an environment which simulated microgravity

    Science.gov (United States)

    Lawless, Brother Desales

    1990-01-01

    Previous research indicated that mouse bone marrow cells could be grown in conditions of simulated microgravity. This environment was created in rotating bioreactor vessels. On three attempts mouse cells were grown successfully in the vessels. The cells reached a stage where the concentrations were doubling daily. Phenotypic analysis using a panel of monoclonal antibodies indicated that the cell were hematopoietic pluripotent stem cells. One unsuccessful attempt was made to reestablish the immune system in immunocompromised mice using these cells. Since last summer, several unsuccessful attempts were made to duplicate these results. It was determined by electron microscopy that the cells successfully grown in 1989 contained virus particles. It was suggested that these virally parasitized cells had been immortalized. The work of this summer is a continuation of efforts to grow mouse bone marrow in these vessels. A number of variations of the protocol were introduced. Certified pathogen free mice were used in the repeat experiments. In some attempts the medium of last summer was used; in others Dexture Culture Medium containing Iscove's Medium supplemented with 20 percent horse serum and 10-6 M hydrocortisone. Efforts this summer were directed solely to repeating the work of last summer. Plans were made for investigations if stem cells were isolated. Immortalization of the undifferentiated stem cell would be attempted by transfection with an oncogenic vector. Selective differentiation would be induced in the stem cell line by growing it with known growth factors and immune response modulators. Interest is in identifying any surface antigens unique to stem cells that would help in their characterization. Another goal was to search for markers on stem cells that would distinguish them from stem cells committed to a particular lineage. If the undifferentiated hematopoietic stem cell was obtained, the pathways that would terminally convert it to myeloid, lyphoid

  5. Modeling and simulation of xylitol production in bioreactor by Debaryomyces nepalensis NCYC 3413 using unstructured and artificial neural network models.

    Science.gov (United States)

    Pappu, J Sharon Mano; Gummadi, Sathyanarayana N

    2016-11-01

    This study examines the use of unstructured kinetic model and artificial neural networks as predictive tools for xylitol production by Debaryomyces nepalensis NCYC 3413 in bioreactor. An unstructured kinetic model was proposed in order to assess the influence of pH (4, 5 and 6), temperature (25°C, 30°C and 35°C) and volumetric oxygen transfer coefficient kLa (0.14h(-1), 0.28h(-1) and 0.56h(-1)) on growth and xylitol production. A feed-forward back-propagation artificial neural network (ANN) has been developed to investigate the effect of process condition on xylitol production. ANN configuration of 6-10-3 layers was selected and trained with 339 experimental data points from bioreactor studies. Results showed that simulation and prediction accuracy of ANN was apparently higher when compared to unstructured mechanistic model under varying operational conditions. ANN was found to be an efficient data-driven tool to predict the optimal harvest time in xylitol production.

  6. Dynamic mathematical models for biodegradation of formaldehyde by Ralstonia eutropha in a batch bioreactor.

    Science.gov (United States)

    Habibi, Alireza; Vahabzadeh, Farzaneh; Zaiat, Marcelo

    2013-11-15

    Degradation of formaldehyde by Ralstonia eutropha was studied in a batch bioreactor operated in recycling mode (30 °C, initial pH of 6.5, aeration rate 0.5 vvm, and a recycling flow rate of 6 mL min(-1)). Growth kinetics equations were described using four substrate inhibition models, and the initial formaldehyde concentration ranged from 54.5 to 993.0 mg L(-1). In each case, model parameters were estimated interactively using nonlinear regression analysis and on the basis of the goodness of fit, the fitness of the model to the experimental data was obtained (i.e., the coefficient of determination and the percent of standard deviation). The estimated parameters according to the Luong equation were μmax = 0.101 h(-1), KS = 54.1 mg L(-1), Sm = 1329 mg L(-1), and n = 2.07. According to the maintenance energies explained by Pirt, cell maintenance was quantified with q = Aμ + B; where A and B are the associated and non-associated growth parts of substrate consumption, respectively. The importance of these terms was verified using the developed models, which would efficiently describe the dynamic nature of growth and formaldehyde biodegradation.

  7. Modelling the biological performance of a side-stream membrane bioreactor using ASM1

    Institute of Scientific and Technical Information of China (English)

    TIAN Ke-jun; LIU Xin-ai; JIANG Tao; M.D. Kennedy; J.C. Schippers; P.A. Vanrolleghem

    2004-01-01

    Membrane bioreactors(MBRs) are attracting global interest but the mathematical modeling of the biological performance of MBRs remains very limited. This study focuses on the modelling of a side-stream MBR system using Activated Sludge Model No.1(ASM1), and comparing the results with the modelling of traditional activated sludge processes. ASM1 parameters relevant for the long-term biological behaviour in MBR systems were calibrated(i.e. YH=0.72gCOD/gCOD, YA=0.25gCOD/gN, bH=0.25 d-1, bA=0.080 d-1 and fP=0.06), and generally agreed with the parameters in traditional activated sludge processes, with the exception that a higher autotrophic biomass decay rate was observed in the MBR. A sensitivity analysis for steady state operation and DO dynamics suggested that the biological performance of the MBR system(the sludge concentration, effluent quality and the DO dynamics) are very sensitive to the parameters(i.e. YH, YA, bH, bA, (maxH and (maxA), and influent wastewater components(XI, Ss, Xs, SNH).

  8. Production of biomass and bioactive compounds from adventitious roots by optimization of culturing conditions of Eurycoma longifolia in balloon-type bubble bioreactor system.

    Science.gov (United States)

    Lulu, Tao; Park, So-Young; Ibrahim, Rusli; Paek, Kee-Yoeup

    2015-06-01

    The present study aimed to optimize the conditions for the production of adventitious roots from Eurycoma longifolia Jack, an important medicinal woody plant, in bioreactor culture. The effects of the type and concentration of auxin on root growth were studied, as well as the effects of the NH4(+):NO3(-) ratio on adventitious root growth and the production of phenolics and flavonoids. Approximately 5 g L(-1) fresh weight of adventitious roots was inoculated into a 3 L balloon-type bubble bioreactor, which contained 2 L 3/4 MS medium supplemented with 30 g L(-1) sucrose and cultures were maintained in the dark for 7 weeks at 24 ± 1°C. Higher concentrations of IBA (7.0 and 9.0 mg L(-1)) and NAA (5.0 mg L(-1)) enhanced the biomass and accumulation of total phenolics and flavonoids. The adventitious roots were thin, numerous, and elongated in 3/4 MS medium supplemented with 5.0 and 7.0 mg L(-1) IBA, whereas the lateral roots were shorter and thicker with 5.0 mg L(-1) NAA compared with IBA treatment. The optimum biomasses of 50.22 g L(-1) fresh weight and 4.60 g L(-1) dry weight were obtained with an NH4(+):NO3(-) ratio of 15:30. High phenolic and flavonoid productions (38.59 and 11.27 mg L(-1) medium, respectively) were also obtained with a ratio of 15:30. Analysis of the 2,2-diphenyl-1-picrylhydrazyl (DPPH)-scavenging activity indicated higher antioxidant activity with an NH4(+):NO3(-) ratio of 30:15. These results suggest that balloon-type bubble bioreactor cultures are suitable for the large-scale commercial production of E. longifolia adventitious roots which contain high yield of bioactive compounds.

  9. Temperature limited fed-batch technique for control of proteolysis in Pichia pastoris bioreactor cultures

    Directory of Open Access Journals (Sweden)

    Garcia Percival

    2003-06-01

    Full Text Available Abstract Background A temperature limited fed-batch (TLFB technique is described and used for Pichia pastoris Mut+ strain cultures and compared with the traditional methanol limited fed-batch (MLFB technique. A recombinant fusion protein composed of a cellulose-binding module (CBM from Neocallimastix patriciarum cellulase 6A and lipase B from Candida antarctica (CALB, was produced and secreted by this strain. Results A protein concentration of about 1 g L-1 was produced in the MLFB process. However, this product was considerably degraded by protease(s. By applying the TLFB process, the yield was increased to 2 g L-1 full-length product and no proteolytic degradation was observed. Flow cytometry analysis showed that the percentage of dead cells increased rapidly during the initial methanol feed phase in the MLFB process and reached a maximum of about 12% after about 40–70 hours of methanol feeding. In the TLFB process, cell death rate was low and constant and reached 4% dead cells at the end of cultivation (about 150 hours methanol feeding time. The lower cell death rate in the TLFB correlated with a lower protease activity in the culture supernatant. The specific alcohol oxidase (AOX activity in the TLFB process was 3.5 times higher than in the MLFB process. Conclusion Three mechanisms that may contribute to the much higher accumulation of product in the TLFB process are: 1 reduced proteolysis due to lower temperature, 2 reduced proteolysis due to lower cell death and protease release to the medium, 3 increased synthesis rate due to higher AOX activity.

  10. T2LBM Version 1.0: Landfill bioreactor model for TOUGH2

    Energy Technology Data Exchange (ETDEWEB)

    Oldenburg, Curtis M.

    2001-05-22

    The need to control gas and leachate production and minimize refuse volume in landfills has motivated the development of landfill simulation models that can be used by operators to predict and design optimal treatment processes. T2LBM is a module for the TOUGH2 simulator that implements a Landfill Bioreactor Model to provide simulation capability for the processes of aerobic or anaerobic biodegradation of municipal solid waste and the associated flow and transport of gas and liquid through the refuse mass. T2LBM incorporates a Monod kinetic rate law for the biodegradation of acetic acid in the aqueous phase by either aerobic or anaerobic microbes as controlled by the local oxygen concentration. Acetic acid is considered a proxy for all biodegradable substrates in the refuse. Aerobic and anaerobic microbes are assumed to be immobile and not limited by nutrients in their growth. Methane and carbon dioxide generation due to biodegradation with corresponding thermal effects are modeled. The numerous parameters needed to specify biodegradation are input by the user in the SELEC block of the TOUGH2 input file. Test problems show that good matches to laboratory experiments of biodegradation can be obtained. A landfill test problem demonstrates the capabilities of T2LBM for a hypothetical two-dimensional landfill scenario with permeability heterogeneity and compaction.

  11. Model-based energy optimisation of a small-scale decentralised membrane bioreactor for urban reuse.

    Science.gov (United States)

    Verrecht, Bart; Maere, Thomas; Benedetti, Lorenzo; Nopens, Ingmar; Judd, Simon

    2010-07-01

    The energy consumption of a small-scale membrane bioreactor, treating high strength domestic wastewater for community level wastewater recycling, has been optimised using a dynamic model of the plant. ASM2d was chosen as biological process model to account for the presence of phosphate accumulating organisms. A tracer test was carried out to determine the hydraulic behaviour of the plant. To realistically simulate the aeration demand, a dedicated aeration model was used incorporating the dependency of the oxygen transfer on the mixed liquor concentration and allowing differentiation between coarse and fine bubble aeration, both typically present in MBRs. A steady state and dynamic calibration was performed, and the calibrated model was able to predict effluent nutrient concentrations and MLSS concentrations accurately. A scenario analysis (SCA) was carried out using the calibrated model to simulate the effect of varying SRT, recirculation ratio and DO set point on effluent quality, MLSS concentrations and aeration demand. Linking the model output with empirically derived correlations for energy consumption allowed an accurate prediction of the energy consumption. The SCA results showed that decreasing membrane aeration and SRT were most beneficial towards total energy consumption, while increasing the recirculation flow led to improved TN removal but at the same time also deterioration in TP removal. A validation of the model was performed by effectively applying better operational parameters to the plant. This resulted in a reduction in energy consumption by 23% without compromising effluent quality, as was accurately predicted by the model. This modelling approach thus allows the operating envelope to be reliably identified for meeting criteria based on energy demand and specific water quality determinants.

  12. Novel perfused compression bioreactor system as an in vitro model to investigate fracture healing

    Directory of Open Access Journals (Sweden)

    Waldemar eHoffmann

    2015-02-01

    Full Text Available Secondary bone fracture healing is a physiological process that leads to functional tissue regeneration via endochondral bone formation. In vivo studies have demonstrated that early mobilization and the application of mechanical loads enhances the process of fracture healing. However, the influence of specific mechanical stimuli and particular effects during specific phases of fracture healing remain to be elucidated. In this work, we have developed and provided proof-of-concept of an in vitro human organotypic model of physiological loading of a cartilage callus, based on a novel perfused compression bioreactor system (PCB. We then used the fracture callus model to investigate the regulatory role of dynamic mechanical loading. Our findings provide a proof-of-principle that dynamic mechanical loading applied by the PCB can enhance the maturation process of mesenchymal stromal cells towards late hypertrophic chondrocytes and the mineralization of the deposited extracellular matrix. The PCB provides a promising tool to study fracture healing and for the in vitro assessment of alternative fracture treatments based on engineered tissue grafts or pharmaceutical compounds, allowing for the reduction of animal experiments.

  13. Study on enhanced denitrification using particulate organic matter in membrane bioreactor by mechanism modeling.

    Science.gov (United States)

    Zheng, Min; Liu, Yan-Chen; Wang, Cheng-Wen; Xu, Kang-Ning

    2013-11-01

    Particulate organic matter (POM) in wastewater is a potential denitrification carbon source, while the optimal operational mode using denitrification mechanism with POM is still unclear in wastewater treatment plants. In this work, we investigated the denitrification rates (DNRs) in a full-scale membrane bioreactor (MBR) coupled with two-stage pre-anoxic (pre-AN), and then evaluated the POM denitrification efficiency using mechanism modeling. The results indicate that POM related fraction accounted for the majority of the obtained specific DNR of 1.39±0.46mgNg(-1) MLVSS h(-1) in the second pre-AN without available soluble carbon source. The modeling approaches with calibration and validation procedures estimated a high residual POM concentration of 0.17g COD g(-1) MLVSS in the activated sludge, which provided specific DNR of 1.14mgNg(-1) MLVSS h(-1). High POM retention time in the reactor was the result of high solid retention time used in the MBR. In particular, post-AN of high biomass concentration could provide the highest POM denitrification efficiency in MBR. The MBR process combined with additional sludge reduction technology could further enhance denitrification by POM.

  14. Comparative modeling of biological nutrient removal from landfill leachate using a circulating fluidized bed bioreactor (CFBBR).

    Science.gov (United States)

    Eldyasti, Ahmed; Andalib, Mehran; Hafez, Hisham; Nakhla, George; Zhu, Jesse

    2011-03-15

    Steady state operational data from a pilot scale circulating fluidized bed bioreactor (CFBBR) during biological treatment of landfill leachate, at empty bed contact times (EBCTs) of 0.49, and 0.41 d and volumetric nutrients loading rates of 2.2-2.6 kg COD/(m(3)d), 0.7-0.8 kg N/(m(3)d), and 0.014-0.016 kg P/(m(3)d), was used to calibrate and compare developed process models in BioWin(®) and AQUIFAS(®). BioWin(®) and AQUIFAS(®) were both capable of predicting most of the performance parameters such as effluent TKN, NH(4)-N, NO(3)-N, TP, PO(4)-P, TSS, and VSS with an average percentage error (APE) of 0-20%. BioWin(®) underpredicted the effluent BOD and SBOD values for various runs by 80% while AQUIFAS(®) predicted effluent BOD and SBOD with an APE of 50%. Although both calibrated models, confirmed the advantages of the CFBBR technology in treating the leachate of high volumetric loading and low biomass yields due to the long solid retention time (SRT), both BioWin(®) and AQUIFAS(®) predicted the total biomass and SRT of CFBBR based on active biomass only, whereas in the CFBBR runs both active as well as inactive biomass accumulated.

  15. Artificial neural network based modeling to evaluate methane yield from biogas in a laboratory-scale anaerobic bioreactor.

    Science.gov (United States)

    Nair, Vijay V; Dhar, Hiya; Kumar, Sunil; Thalla, Arun Kumar; Mukherjee, Somnath; Wong, Jonathan W C

    2016-10-01

    The performance of a laboratory-scale anaerobic bioreactor was investigated in the present study to determine methane (CH4) content in biogas yield from digestion of organic fraction of municipal solid waste (OFMSW). OFMSW consists of food waste, vegetable waste and yard trimming. An organic loading between 40 and 120kgVS/m(3) was applied in different runs of the bioreactor. The study was aimed to focus on the effects of various factors, such as pH, moisture content (MC), total volatile solids (TVS), volatile fatty acids (VFAs), and CH4 fraction on biogas production. OFMSW witnessed high CH4 yield as 346.65LCH4/kgVS added. A target of 60-70% of CH4 fraction in biogas was set as an optimized condition. The experimental results were statistically optimized by application of ANN model using free forward back propagation in MATLAB environment.

  16. Integrated ‘omics analysis for studying the microbial community response to a pH perturbation of a cellulose-degrading bioreactor culture

    Energy Technology Data Exchange (ETDEWEB)

    Boaro, Amy A. [Washington State Univ., Richland, WA (United States). Bioproducts, Sciences and Engineering Lab.; Kim, Young-Mo [Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Biological Sciences Division; Konopka, Allan E. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Biological Sciences Division; Callister, Stephen J. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Biological Sciences Division; Ahring, Birgitte K. [Washington State Univ., Richland, WA (United States). Bioproducts, Sciences and Engineering Lab.

    2015-01-05

    Propionate accumulation is a common indicator of process imbalances in anaerobic bioreactor systems. The accumulation of propionate can occur due to low retention rates, hydrogen accumulation, or mechanical changes affecting the proximity between propionate oxidizers and partner species, thereby preventing necessary electron transfer. Few studies, however, have observed the changes in microbial community structure during propionate accumulation. We used 454 pyrosequencing of 16S rDNA to evaluate the community membership during propionate accumulations in replicate bioreactors with rumen based cultures. Half of the culture volume from a parent reactor was transferred to a sterile “daughter” reactor, and both systems were run identically. Both reactors experienced a propionate accumulation after roughly 10 days, with the propionate accumulation being less pronounced in the parent reactor as compared to the daughter reactor. Non-metric multidimensional scaling (NMDS) was used to determine clustering patterns of the samples, and correlative methods were used to determine which OTUs were significantly associated with the movements of samples along the NMDS axes. The presence of Saccharofermentans characterized the position of early samples, whereas the presence of Ruminococcus and Succiniclasticum were more indicative of the positions of later samples. Hydrogen accumulation and low sequence counts indicated low methanogen activity. Although both reactor systems were closed to microbial inputs due to the sterilization of influent media, we recorded significant increases in reactor diversity over time. This suggests that changes in the abundances of dominant community members may affect the sequencing of rare taxa within samples.

  17. Challenges of culturing human norovirus in three-dimensional organoid intestinal cell culture models.

    Directory of Open Access Journals (Sweden)

    Efstathia Papafragkou

    Full Text Available Human noroviruses are the most common cause of acute gastroenteritis worldwide. Recently, cell culture systems have been described using either human embryonic intestinal epithelial cells (Int-407 or human epithelial colorectal adenocarcinoma cells (Caco-2 growing on collagen-I porous micro carrier beads in a rotating bioreactor under conditions of physiological fluid shear. Here, we describe the efforts from two independent laboratories to implement this three dimensional (3D cell culture system for the replication of norovirus. Int-407 and Caco-2 were grown in a rotating bioreactor for up to 28 days. Prior to infection, cells were screened for the presence of microvilli by electron microscopy and stained for junction proteins (zonula occludens-1, claudin-1, and β-catenin. Differentiated 3D cells were transferred to 24-well plates and infected with bacteria-free filtrates of various norovirus genotypes (GI.1, GI.3, GI.8, GII.2, GII.4, GII.7, and GII.8. At 12 h, 24 h, and 48 h post inoculation, viral RNA from both cells and supernatants were collected and analyzed for norovirus RNA by real-time reverse transcription PCR. Despite observations of high expression of junction proteins and microvilli development in stained thin sections, our data suggest no significant increase in viral titer based on norovirus RNA copy number during the first 48 h after inoculation for the different samples and virus culture conditions tested. Our combined efforts demonstrate that 3D cell culture models using Int-407 or Caco-2 cells do not support norovirus replication and highlight the complexity and difficulty of developing a reproducible in vitro cell culture system for human norovirus.

  18. Model-based cell number quantification using online single-oxygen sensor data for tissue engineering perfusion bioreactors.

    Science.gov (United States)

    Lambrechts, T; Papantoniou, I; Sonnaert, M; Schrooten, J; Aerts, J-M

    2014-10-01

    Online and non-invasive quantification of critical tissue engineering (TE) construct quality attributes in TE bioreactors is indispensable for the cost-effective up-scaling and automation of cellular construct manufacturing. However, appropriate monitoring techniques for cellular constructs in bioreactors are still lacking. This study presents a generic and robust approach to determine cell number and metabolic activity of cell-based TE constructs in perfusion bioreactors based on single oxygen sensor data in dynamic perfusion conditions. A data-based mechanistic modeling technique was used that is able to correlate the number of cells within the scaffold (R(2)  = 0.80) and the metabolic activity of the cells (R(2)  = 0.82) to the dynamics of the oxygen response to step changes in the perfusion rate. This generic non-destructive measurement technique is effective for a large range of cells, from as low as 1.0 × 10(5) cells to potentially multiple millions of cells, and can open-up new possibilities for effective bioprocess monitoring.

  19. A computational model for the optimization of transport phenomena in a rotating hollow-fiber bioreactor for artificial liver.

    Science.gov (United States)

    Consolo, Filippo; Fiore, Gianfranco B; Truscello, Silvia; Caronna, Marco; Morbiducci, Umberto; Montevecchi, Franco M; Redaelli, Alberto

    2009-03-01

    A comprehensive computational study modelling the operation of a rotating hollow-fiber bioreactor for artificial liver (BAL) was performed to explore the interactions between the oxygenated culture medium and the cultured hepatocytes. Computational fluid dynamics investigations were carried out using two-dimensional (2D) and 3D time-dependent numerical simulations, integrating calculations of diffusion, convection, and multiphase fluid dynamics. The analysis was aimed at determining the rotational speed value of the chamber to ensure homogenous distribution of the floating microcarrier-attached aggregated cells (microCAACs) and avoid their sedimentation and excessive packing, analyzing oxygen (O(2)) delivery and cellular O(2) consumption as an index of cellular metabolic activity, and analyzing the fluid-induced mechanical stress experienced by cells. According to our results, homogeneous distribution of cells is reached at a rotational speed of 30 rpm; spreading of cellular concentration at around the initial value of 12% was limited (median = 11.97%, 5th percentile = 10.94%, 95th percentile = 13.2%), resulting in uniform suspension of microCAACs, which did not appear to be excessively packed. Mixing within the rotating fluid caused a maximum fluid-induced stress value of 0.05 Pa, which was neither endangering for liver-specific functions of cultured cells, nor causing disruption of the floating aggregates. Moreover, an inlet medium flow rate of 200 mL/m with a partial pressure of oxygen (pO(2)) value of 160 mmHg was found to guarantee an adequate O(2) supply for the hepatocytes (2.7 x 10(8) hepatocytes are simulated); under such conditions, the minimum pO(2) value (23 mmHg) is above the critical threshold value, causing the onset of cellular hypoxia (10 mmHg). We proved that numerical simulation of transport phenomena is a valuable tool for the computer-aided design of BALs, helping overcome the unsolved issues in optimizing the cell-environment conditioning

  20. Analysis of drug metabolism activities in a miniaturized liver cell bioreactor for use in pharmacological studies.

    Science.gov (United States)

    Hoffmann, Stefan A; Müller-Vieira, Ursula; Biemel, Klaus; Knobeloch, Daniel; Heydel, Sandra; Lübberstedt, Marc; Nüssler, Andreas K; Andersson, Tommy B; Gerlach, Jörg C; Zeilinger, Katrin

    2012-12-01

    Based on a hollow fiber perfusion technology with internal oxygenation, a miniaturized bioreactor with a volume of 0.5 mL for in vitro studies was recently developed. Here, the suitability of this novel culture system for pharmacological studies was investigated, focusing on the model drug diclofenac. Primary human liver cells were cultivated in bioreactors and in conventional monolayer cultures in parallel over 10 days. From day 3 on, diclofenac was continuously applied at a therapeutic concentration (6.4 µM) for analysis of its metabolism. In addition, the activity and gene expression of the cytochrome P450 (CYP) isoforms CYP1A2, CYP2B6, CYP2C9, CYP2D6, and CYP3A4 were assessed. Diclofenac was metabolized in bioreactor cultures with an initial conversion rate of 230 ± 57 pmol/h/10(6) cells followed by a period of stable conversion of about 100 pmol/h/10(6) cells. All CYP activities tested were maintained until day 10 of bioreactor culture. The expression of corresponding mRNAs correlated well with the degree of preservation. Immunohistochemical characterization showed the formation of neo-tissue with expression of CYP2C9 and CYP3A4 and the drug transporters breast cancer resistance protein (BCRP) and multidrug resistance protein 2 (MRP2) in the bioreactor. In contrast, monolayer cultures showed a rapid decline of diclofenac conversion and cells had largely lost activity and mRNA expression of the assessed CYP isoforms at the end of the culture period. In conclusion, diclofenac metabolism, CYP activities and gene expression levels were considerably more stable in bioreactor cultures, making the novel bioreactor a useful tool for pharmacological or toxicological investigations requiring a highly physiological in vitro representation of the liver.

  1. Optimization of HEK-293S cell cultures for the production of adenoviral vectors in bioreactors using on-line OUR measurements.

    Science.gov (United States)

    Gálvez, J; Lecina, M; Solà, C; Cairó, J J; Gòdia, F

    2012-01-01

    The culture of HEK-293S cells in a stirred tank bioreactor for adenoviral vectors production for gene therapy is studied. Process monitoring using oxygen uptake rate (OUR) was performed. The OUR was determined on-line by the dynamic method, providing good information of the process evolution. OUR enabled cell activity monitoring, facilitating as well the determination of the feeding rate in perfusion cultures and when to infect the culture. Batch cultures were used to validate the monitoring methodology. A cell density of 10×10(5)cell/mL was infected, producing 1.3×10(9) infectious viral particles/mL (IVP/mL). To increase cell density values maintaining cell specific productivity, perfusion cultures, based on tangential flow filtration, were studied. In this case, OUR measurements were used to optimize the dynamic culture medium feeding strategy, addressed to avoid any potential nutrient limitation. Furthermore, the infection protocol was defined in order to optimize the use of the viral inoculum, minimizing the uncontrolled release of particles through the filter unit mesh. All these developments enabled an infection at 78×10(5)cell/mL with the consequent production of 44×10(9)IVP/mL, representing a cell specific productivity 4.3 times higher than for the batch culture.

  2. Modelling and Extremum Seeking Control of a Cascade of Two Anaerobic Bioreactors

    Directory of Open Access Journals (Sweden)

    Ivan Simeonov

    2011-05-01

    Full Text Available The principle of extremum seeking control has been applied on a cascade of two anaerobic bioreactors using the dilution rate as control action and the biogas flow rates as measured outputs to be maximized. In all cases maximum biogas flow rate with sensible decrease of the general output depollution parameter (compared to the case of one single bioreactor has been obtained, starting from different initial conditions. With the same algorithm, good performances have been obtained in the presence of variations of the inlet organics. Its implication for biotechnology may result in substantial economic benefits.

  3. Tissue grown in NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    Cells from kidneys lose some of their special features in conventional culture but form spheres replete with specialized cell microvilli (hair) and synthesize hormones that may be clinically useful. Ground-based research studies have demonstrated that both normal and neoplastic cells and tissues recreate many of the characteristics in the NASA bioreactor that they display in vivo. Proximal kidney tubule cells that normally have rich apically oriented microvilli with intercellular clefts in the kidney do not form any of these structures in conventional two-dimensional monolayer culture. However, when normal proximal renal tubule cells are cultured in three-dimensions in the bioreactor, both the microvilli and the intercellular clefts form. This is important because, when the morphology is recreated, the function is more likely also to be rejuvenated. 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).

  4. Tissue grown in NASA Bioreactor

    Science.gov (United States)

    1998-01-01

    Cells from kidneys lose some of their special features in conventional culture but form spheres replete with specialized cell microvilli (hair) and synthesize hormones that may be clinically useful. Ground-based research studies have demonstrated that both normal and neoplastic cells and tissues recreate many of the characteristics in the NASA bioreactor that they display in vivo. Proximal kidney tubule cells that normally have rich apically oriented microvilli with intercellular clefts in the kidney do not form any of these structures in conventional two-dimensional monolayer culture. However, when normal proximal renal tubule cells are cultured in three-dimensions in the bioreactor, both the microvilli and the intercellular clefts form. This is important because, when the morphology is recreated, the function is more likely also to be rejuvenated. 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).

  5. Bioreactors for Plant Embryogenesis and Beyond.

    Science.gov (United States)

    Fei, Liwen; Weathers, Pamela

    2016-01-01

    A variety of different bioreactors have been developed for use in initiating and cultivating somatic embryos. The various designs for embryogenesis and culture are critically evaluated here. Bioreactor optimization and operation methods are also described along with recommendations for use based on desired outcome.

  6. Numerical Simulation of Microcarrier Motion in a Rotating Wall Vessel Bioreactor

    Institute of Scientific and Technical Information of China (English)

    ZHI-HAO JU; TIAN-QING LIU; XUE-HU MA; ZHAN-FENG CUI

    2006-01-01

    Objective To analyze the forces of rotational wall vessel (RWV) bioreactor on small tissue pieces or microcarrier particles and to determine the tracks of microcarrier particles in RWV bioreactor. Methods The motion of the microcarrier in the rotating wall vessel (RWV) bioreactor with both the inner and outer cylinders rotating was modeled by numerical simulation. Results The continuous trajectory of microcarrier particles, including the possible collision with the wall was obtained. An expression between the minimum rotational speed difference of the inner and outer cylinders and the microcarrier particle or aggregate radius could avoid collisions with either wall. The range of microcarrier radius or tissue size, which could be safely cultured in the RWV bioreactor, in terms of shear stress level, was determined. Conclusion The model works well in describing the trajectory of a heavier microcarrier particle in rotating wall vessel.

  7. DESIGNING SULFATE-REDUCING BACTERIA FIELD-BIOREACTORS USING THE BEST MODEL

    Science.gov (United States)

    DESIGNING SULFATE-REDUCING BACTERIA FIELD-BIOREACTORS USING THE BEST MODELMarek H. Zaluski1,3, Brian T. Park1, Diana R. Bless21 MSE Technology Applications; 200 Technology Way, Butte, Montana 59701, USA2 U.S. EPA, Office of Research and Development, Cincinna...

  8. Application of microgravity bioreactor in cell culture%微重力反应器在细胞培养中的应用

    Institute of Scientific and Technical Information of China (English)

    季晓昕; 李非; 李铎

    2006-01-01

    目的:微重力培养技术是一种新兴的培养技术,可用于组织工程、细胞培养等多个领域.介绍微重力反应器模拟微重力环境用于细胞培养.资料来源:应用计算机检索PubMed 1995-01/2002-06的文章,检索词为"microgravity,simulated microgravity,bioreactor,tissue engineering,cellculture,Rotating cell culture system",限定文章语言种类为English.资料选择:纳入标准:①模拟微重力下细胞培养的研究.②微重力组织工程学的研究.③随机对照实验研究.④微重力环境的创建.排除标准:①较陈旧的文献.②重复研究.资料提炼:共收集到288篇与脊髓损伤修复相关的文献,其中19篇符合纳入标准,排除的269篇文章系同一类重复性研究.资料综合:①模拟微重力培养有其独特的特点和实用性:避免湍流形成的过大剪切力;培养液通过旋转逐渐混合;零顶空间;贴壁依赖性细胞可在载体上生长;提供充分氧合作用.②微重力环境下细胞的生长:细胞易于聚集,产生更高的细胞密度.③微重力反应器可以建立动物细胞的三维培养体系,达到满足器官替代物,或体外研究的要求.④通过微重力反应器所构建的三维组织结构中可产生血管样管状结构,用于产生组织工程学的血管管道.⑤需要解决的问题:微重力反应器所构建的三维结构一旦离开微重力环境,其功能能否长期保持,或移植整合到宿主组织后仍保持不变,尚需进一步研究.结论:模拟微重力培养是一种全新的组织培养技术,其核心技术是建立动物细胞的三维培养体系,可以为多种细胞和组织块的生长和代谢提供良好的培养环境,可以进行高密度的组织培养,并保持所培养细胞的组织分化特异性.%OBJECTIVE: Microgravity culture is an emerging cultural technology,which can be used in many fields such as tissue engineering and cell culture,etc. Microgravity bioreactor was used for cell culture

  9. Perfusion Bioreactor Module

    Science.gov (United States)

    Morrison, Dennis R.

    1990-01-01

    Perfusion bioreactor module, self-contained, closed-loop cell-culture system that operates in microgravity or on Earth. Equipment supports growth or long-term maintenance of cultures of human or other fragile cells for experiments in basic cell biology or process technology. Designed to support proliferation (initially at exponential rates of growth) of cells in complex growth medium and to maintain confluent cells in defined medium under conditions optimized to permit or encourage selected functions of cells, including secretion of products of cells into medium.

  10. Bioreactor landfill

    Institute of Scientific and Technical Information of China (English)

    WANG Hao; XING Kai; Anthony Adzomani

    2004-01-01

    Following the population expansion, there is a growing threat brought by municipal solid waste (MSW) against environment and human health. Sanitary landfill is the most important method of MSW disposal in China. In contrast to the conventional landfill, this paper introduces a new technique named bioreactor landfill (BL). Mechanisms, operation conditions as well as the advantages and disadvantages of BL are also discussed in this paper.

  11. Optimal Homogenization of Perfusion Flows in Microfluidic Bio-Reactors: A Numerical Study

    DEFF Research Database (Denmark)

    Okkels, Fridolin; Dufva, Martin; Bruus, Henrik

    2011-01-01

    In recent years, the interest in small-scale bio-reactors has increased dramatically. To ensure homogeneous conditions within the complete area of perfused microfluidic bio-reactors, we develop a general design of a continually feed bio-reactor with uniform perfusion flow. This is achieved...... by introducing a specific type of perfusion inlet to the reaction area. The geometry of these inlets are found using the methods of topology optimization and shape optimization. The results are compared with two different analytic models, from which a general parametric description of the design is obtained...... and tested numerically. Such a parametric description will generally be beneficial for the design of a broad range of microfluidic bioreactors used for, e. g., cell culturing and analysis and in feeding bio-arrays....

  12. Transgenic bioreactors.

    Science.gov (United States)

    Jänne, J; Alhonen, L; Hyttinen, J M; Peura, T; Tolvanen, M; Korhonen, V P

    1998-01-01

    Since the generation of the first transgenic mice in 1980, transgene technology has also been successfully applied to large farm animals. Although this technology can be employed to improve certain production traits of livestock, this approach has not been very successful so far owing to unwanted effects encountered in the production animals. However, by using tissue-specific targeting of the transgene expression, it is possible to produce heterologous proteins in the extracellular space of large transgenic farm animals. Even though some recombinant proteins, such as human hemoglobin, have been produced in the blood of transgenic pigs, in the majority of the cases mammary gland targeted expression of the transgene has been employed. Using production genes driven by regulatory sequences of milk protein genes a number of valuable therapeutic proteins have been produced in the milk of transgenic bioreactors, ranging from rabbits to dairy cattle. Unlike bacterial fermentors, the mammary gland of transgenic bioreactors appear to carry out proper postsynthetic modifications of human proteins required for full biological activity. In comparison with mammalian cell bioreactors, transgenic livestock with mammary gland targeted expression seems to be able to produce valuable human therapeutic proteins at very low cost. Although not one transgenically produced therapeutic protein is yet on the market, the first such proteins have recently entered or even completed clinical trials required for their approval.

  13. Model studies and numerical simulations on nitrate reduction in a bioreactor; Modellbetrachtung und numerische Simulationen zur Nitrat-Reduktion in einem Bioreaktor

    Energy Technology Data Exchange (ETDEWEB)

    Ghergut, I. [Inst. fuer angew. Math., Rumaenische Akad. d. Wiss. (Romania); Maloszewski, P.; Seiler, K.P. [GSF - Forschungszentrum fuer Umwelt und Gesundheit GmbH, Neuherberg (Germany). Inst. fuer Hydrologie; Naumann, U.

    2001-11-01

    Naumann carried out denitrification studies on bacteria in a bioreactor providing a groundwater-like, anoxic environment which was so adjusted as to sustain a stable, denitrifying biocoenosis. This bioreactor consists of a column containing water from a karst spring and drill core material from the mass lime of the Franconian Jurassic. There is a flow of spring water through the bioreactor. This paper is an endeavour to formulate classical models on nutrient consumption and population dynamics in this bioreactor. The processes in the bioreactor are determined quantitatively by means of simple partial differential equations. This leads to a calibrated mathematical model which serves as a basis for discussing and specifying unresolved questions. [German] Naumann fuehrte Denitrifikationsuntersuchungen von Bakterien in einem unter grundwasseraehnlichen, sauerstoffreien Bedingungen funktionierenden Bioreaktor durch, in dem Voraussetzungen einer stabilen denitrifizierenden Organismengemeinschaft geschaffen wurden. Dieser Bioreaktor besteht aus einer Saeule mit Wasser aus einer Karstquelle und Bohrkernmaterial aus dem Massenkalk des fraenkischen Juras und wird vom Quellwasser durchstroemt. Im Folgenden wird versucht, klassische Modellansaetze zum Naehrstoffverbrauch und zur Populationsdynamik fuer das Geschehen in diesem Bioreaktor zu artikulieren; das Bioreaktorgeschehen wird durch einfache partielle Differential-Gleichungen (PDG) quantitativ erfasst. Das so kalibrierte mathematische Modell wird schliesslich dazu benutzt noch offene Fragen anzudiskutieren bzw. einzuengen. (orig.)

  14. A microcarrier cell culture process for propagating rabies virus in Vero cells grown in a stirred bioreactor under fully animal component free conditions.

    Science.gov (United States)

    Rourou, Samia; van der Ark, Arno; van der Velden, Tiny; Kallel, Héla

    2007-05-10

    Rabies virus strain production in Vero cells grown on Cytodex 1 in a 2 L stirred tank bioreactor and in a medium free of components of human or animal origin (VP-SFM) is described. Cell banking procedure in VP-SFM supplemented with an animal components free mixture (10%DMSO+0.1%methylcellulose) was reported and cell growth after revitalization was assessed. Vero cells exhibited growth performances (specific growth rate and cell division number) similar to that obtained in serum containing medium. To design a scalable process that is totally free of animal-derived substances, two proteases: TrypLE Select and Accutase, were assessed as an alternative to trypsin which is routinely used for cell passage. Growth performance of Vero cells grown in VP-SFM and MEM+10% fetal calf serum (FCS) over four passages and subcultivated with either TrypLE Select or Accutase was evaluated. TrypLE Select showed the best performance in terms of specific growth rate and cell division number. Kinetics of cell growth and rabies virus production (LP2061/Vero strain) were investigated in spinner flask and in a 2 L bioreactor. In spinner flask, a maximal cell density level of 1.85x10(6) cells/mL was achieved when the cells were grown in VP-SFM on 2 g/L Cytodex 1. Cell infection experiments conducted at an MOI of 0.3 and without the medium exchange step, typically needed for serum containing rabies virus production, resulted in a maximal virus titer equal to 2x10(7) (Fluorescent Focus Unit) FFU/mL. In stirred tank bioreactor, Vero cell growth in VP-SFM on 3 g/L Cytodex 1 was shown to be sensitive to the aeration mode. Sparging the culture was detrimental for cell growth, whereas cell density level was greatly enhanced when only headspace aeration was used. A cell density level of 2.6x10(6) cells/mL was obtained when the cells were grown on 3g/L Cytodex 1 and in batch culture mode. Cell infection at an MOI of 0.1 without any medium exchange, yielded a maximal rabies virus titer of 2.4x10

  15. Concentrations monitoring via software sensor for bioreactors under model parametric uncertainty: Application to cadmium removal in an anaerobic pro

    Directory of Open Access Journals (Sweden)

    P.A. López Pérez

    2016-06-01

    Full Text Available The main objective of this work was to design a software sensor device based on state observer for a class of continuous bioreactor with application to heavy metal removal and locally analyze the observability properties of the considered system, considering parametric uncertainties. First, an alternative phenomenological model of the main state variables of the process was formulated, considering an unstructured kinetic approach based on Levenspiel product inhibition model; this kinetic model was experimentally validated. The kinetic model was used as a benchmark plant and extended for continuous operation in order to analyze the local observability properties, considering several sets of measured outputs that produce observable subspaces of different dimensions. In addition, we present a nonlinear observer, which is robust against parametric uncertainties, to estimate the observable states of the bioreactor. The convergence of the proposed methodology was analyzed using Lyapunov stability theory. Numerical experiments were done in order to show the performance of the proposed observer and the observability properties of the system.

  16. Computational Modeling of Culture's Consequences

    NARCIS (Netherlands)

    Hofstede, G.J.; Jonker, C.M.; Verwaart, T.

    2010-01-01

    This paper presents an approach to formalize the influence of culture on the decision functions of agents in social simulations. The key components are (a) a definition of the domain of study in the form of a decision model, (b) knowledge acquisition based on a dimensional theory of culture, resulti

  17. Modeling of gas-liquid mass transfer in a stirred tank bioreactor agitated by a Rushton turbine or a new pitched blade impeller.

    Science.gov (United States)

    Gelves, Ricardo; Dietrich, A; Takors, Ralf

    2014-03-01

    A combined computational fluid dynamics (CFD) and population balance model (PBM) approach has been applied to simulate hydrodynamics and mass transfer in a 0.18 m(3) gas-liquid stirred bioreactor agitated by (1) a Rushton turbine, and (2) a new pitched blade geometry with rotating cartridges. The operating conditions chosen were motivated by typical settings used for culturing mammalian cells. The effects of turbulence, rotating flow, bubbles breakage and coalescence were simulated using the k-ε, multiple reference frame (MRF), Sliding mesh (SM) and PBM approaches, respectively. Considering the new pitched blade geometry with rotating aeration microspargers, [Formula: see text] mass transfer was estimated to be 34 times higher than the conventional Rushton turbine set-up. Notably, the impeller power consumption was modeled to be about 50 % lower. Independent [Formula: see text] measurements applying the same operational conditions confirmed this finding. Motivated by these simulated and experimental results, the new aeration and stirring device is qualified as a very promising tool especially useful for cell culture applications which are characterized by the challenging problem of achieving relatively high mass transfer conditions while inserting only low stirrer energy.

  18. Influence of inoculum density and aeration volume on biomass and bioactive compound production in bulb-type bubble bioreactor cultures of Eleutherococcus koreanum Nakai.

    Science.gov (United States)

    Lee, Eun-Jung; Moh, Sang-Hyun; Paek, Kee-Yoeup

    2011-07-01

    This study deals with the effects of initial inoculum density and aeration volume on biomass and bioactive compound production in adventitious roots of Eleutherococcus koreanum Nakai in bulb-type bubble bioreactors (3-L capacity). While the fresh and dry weights of the roots increased with increasing inoculum density, the highest percentage dry weight and accumulation of total target compounds (eleutheroside B and E, chlorogenic acid, total phenolics, and flavonoids) were noted at an inoculum density of 5.0 g L(-1). Poor aeration volume (0.05 vvm) stunted root growth, and high aeration volume (0.4 vvm) caused physiological disorders. Moreover, an inoculum density of 5.0 g L(-1) and an aeration volume of 0.1 vvm resulted in the highest concentration of total target compounds and least root death. Such optimization of culture conditions will be beneficial for the large-scale production of E. koreanum biomass and bioactive compounds.

  19. Comparison of flat photo-bioreactors for micro-algae culture based on CFD numerical simulation%基于CFD数值模拟的平板式微藻光生物反应器比较

    Institute of Scientific and Technical Information of China (English)

    齐祥明; 崔海龙

    2015-01-01

    为进一步提高微藻光生物反应器的混合与传质性能,在已有多节隔板平板式光生物反应器的基础上设计多级进气,新建立了多级进气多级隔板平板式光生物反应器。构建了普通反应器、多节隔板反应器、多级进气反应器并利用计算流体动力学模拟研究了3种反应器的流动与传质特性。结果表明,模拟结果与相关试验测量值吻合良好,多级进气结构可以带来更明显的级内环流现象,从而使该反应器在液体平均速度、死区比、湍动能、湍动能耗散率、气含率、液相传质系数等性能参数上较前2种反应器均有很大提高。在适合微藻培养的通气率0.4~0.8(每分钟通入反应器的气体体积与反应器实际装液体积之比)内,该反应器的混合及传质性能均表现优异。该工作为平板式生物反应器的设计及优化提供了新的方向。%Algae as potential resources, has attracted increasing interest and attention from many fields such as energy, medicament, food, feed, and environment. However design and optimization of photo-bioreactor for algae production remains a bottleneck in the development of microalgae culturing industry. Recently, flat photo-bioreactor is improved by changing the double-flat into multistage structure. In this study, in order to further increase mass transfer and mixing properties of microalgae photo-bioreactor, a multistage intake structure was fixed into this multistage flat photo-bioreactor. Moreover, for the purpose of exploring more mass transfer and mixing details of the three photo-bioreactors, the ordinary double-flat photo-bioreactor, multistage flat photo-bioreactor, and multistage intake photo-bioreactor were constructed physically and numerically, and their computational fluids dynamics (CFD) simulations were carried out. Gas holdups and mass transfer coefficients were measured in physical multistage intake photo-bioreactor and compared

  20. Nitrate-Mediated Microbially Enhanced Oil Recovery (N-MEOR) from model upflow bioreactors.

    Science.gov (United States)

    Gassara, Fatma; Suri, Navreet; Voordouw, Gerrit

    2017-02-15

    Microbially Enhanced Oil Recovery (MEOR) can enhance oil production with less energy input and less costs than other technologies. The present study used different aqueous electron donors (acetate, glucose, molasses) and an aqueous electron acceptor (nitrate) to stimulate growth of heterotrophic nitrate reducing bacteria (hNRB) to improve production of oil. Initial flooding of columns containing heavy oil (viscosity of 3400cP at 20°C) with CSBK (Coleville synthetic brine medium) produced 0.5 pore volume (PV) of oil. Bioreactors were then inoculated with hNRB with 5.8g/L of molasses and 0, 10, 20, 40, 60 or 80mM nitrate, as well as with 17mM glucose or 57mM acetate and 80mM nitrate. During incubations no oil was produced in the bioreactors that received 5.8g/L of molasses and 0, 10, 20, 40 or 60mM nitrate. However, the bioreactors injected with 5.8g/L of molasses, 17mM glucose or 57mM acetate and 80mM nitrate produced 13.9, 11.3±3.1 and 17.8±6.6% of residual oil, respectively. The significant production of oil from these bioreactors may be caused by N2-CO2 gas production. Following continued injection with CSBK without nitrate, subsequent elution of significant residual oil (5-30%) was observed. These results also indicate possible involvement of fermentation products (organic acids, alcohols) to enhance heavy oil recovery. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Nutrient removal from an anaerobic membrane bioreactor effluent using microalgae. Study and modeling of the process.

    OpenAIRE

    Ruiz Martínez, Ana

    2016-01-01

    [EN] Anaerobic membrane bioreactors for urban wastewater treatment present interesting advantages when compared with aerobic treatments, such as less sludge production, lower energy demand and biogas generation. However, the generated effluent cannot generally be discharged without further ammonium and phosphate elimination. This thesis studies the removal of these inorganic nutrients by means of microalgae cultivation. The main objective of this work is therefore to obtain an autochthono...

  2. Activated sludge model (ASM) based modelling of membrane bioreactor (MBR) processes: a critical review with special regard to MBR specificities.

    Science.gov (United States)

    Fenu, A; Guglielmi, G; Jimenez, J; Spèrandio, M; Saroj, D; Lesjean, B; Brepols, C; Thoeye, C; Nopens, I

    2010-08-01

    Membrane bioreactors (MBRs) have been increasingly employed for municipal and industrial wastewater treatment in the last decade. The efforts for modelling of such wastewater treatment systems have always targeted either the biological processes (treatment quality target) as well as the various aspects of engineering (cost effective design and operation). The development of Activated Sludge Models (ASM) was an important evolution in the modelling of Conventional Activated Sludge (CAS) processes and their use is now very well established. However, although they were initially developed to describe CAS processes, they have simply been transferred and applied to MBR processes. Recent studies on MBR biological processes have reported several crucial specificities: medium to very high sludge retention times, high mixed liquor concentration, accumulation of soluble microbial products (SMP) rejected by the membrane filtration step, and high aeration rates for scouring purposes. These aspects raise the question as to what extent the ASM framework is applicable to MBR processes. Several studies highlighting some of the aforementioned issues are scattered through the literature. Hence, through a concise and structured overview of the past developments and current state-of-the-art in biological modelling of MBR, this review explores ASM-based modelling applied to MBR processes. The work aims to synthesize previous studies and differentiates between unmodified and modified applications of ASM to MBR. Particular emphasis is placed on influent fractionation, biokinetics, and soluble microbial products (SMPs)/exo-polymeric substances (EPS) modelling, and suggestions are put forward as to good modelling practice with regard to MBR modelling both for end-users and academia. A last section highlights shortcomings and future needs for improved biological modelling of MBR processes.

  3. Submerged Culture of Mushrooms in Bioreactors – Challenges, Current State-of-the-Art, and Future Prospects

    Directory of Open Access Journals (Sweden)

    Li-Wen Zhu

    2007-01-01

    Full Text Available Medicinal mushrooms have profound health-promoting benefits. Recently, a number of substances of mushroom origin have been isolated, identified and shown to have physiological activities, such as antitumor, immunomodulating, cardiovascular, antihypercholesterolemia, antibacterial, antiviral, antiparasitic, hepatoprotective, and antidiabetic activities. Currently, commercial products from medicinal mushrooms are mostly obtained through the field-cultivation of the fruiting body. However, in this case it is difficult to control the quality of the final product. Submerged fermentation of the mycelial form of mushroom-producing fungi has received much attention as a promising alternative for efficient production of the biomass of medicinal mushrooms and their active metabolites. However, in order for the production to be successful at industrial scale, various technical problems need to be solved, including characterization of the variations that occur during the submerged cultivation of mushrooms in bioreactors and their effects on growth and product formation. This review outlines the major factors that affect the submerged cultivation of mushrooms in bioreactors, including oxygen supply, shear and mixing, morphology and rheology, as well as two-stage cultivation strategies and high-cell-density cultivation strategies such as fed-batch fermentation.

  4. Modeling of mixing in stirred bioreactors 4. mixing time for aerated bacteria, yeasts and fungus broths

    Directory of Open Access Journals (Sweden)

    Cascaval Dan

    2004-01-01

    Full Text Available The mixing time for bioreactors depends mainly on the rheoiogicai properties of the broths, the biomass concentration and morphology, mixing system characteristics and fermentation conditions. For quantifying the influence of these factors on the mixing efficiency for stirred bioreactors, aerated broths of bacteria (P. shermanii, yeasts (S. cerevisiae and fungi (P. chrysogenum, free mycelia and mycelial aggregates of different concentrations have been investigated using a laboratory bioreactor with a double turbine impeller. The experimental data indicated that the influence of the rotation speed, aeration rate and stirrer positions on the mixing intensity strongly differ from one system to another and must be correlated with the microorganism characteristics, namely: the biomass concentration and morphology. Moreover, compared with non-aerated broths, variations of the mixing time with the considered parameters are very different, due to the complex flow mechanism of gas-liquid dispersions. By means of the experimental data and using a multiregression analysis method some mathematical correlations for the mixing time of the general form: tm = a1*Cx2+a2*Cx+a3*IgVa+a4-N2+a5-N+a6/a7*L2+a8*L+a9 were established. The proposed equations offer good agreement with the experiments, the average deviation being ±6.7% - ±9.4 and are adequate for the flow regime Re < 25,000.

  5. Development of Fundamental Technologies for Micro Bioreactors

    Science.gov (United States)

    Sato, Kiichi; Kitamori, Takehiko

    This chapter reviews the development of fundamental technologies required for microchip-based bioreactors utilizing living mammalian cells and pressure driven flow. The most important factor in the bioreactor is the cell culture. For proper cell culturing, continuous medium supply from a microfluidic channel and appropriate modification of the channel surface to accommodate cell attachment is required. Moreover, the medium flow rate should be chosen carefully, because shear stress affects cell activity. The techniques presented here could be applied to the development of micro bioreactors such as microlivers, pigment production by plant cells, and artificial insemination.

  6. Improvement of In Vitro Three-Dimensional Cartilage Regeneration by a Novel Hydrostatic Pressure Bioreactor.

    Science.gov (United States)

    Chen, Jie; Yuan, Zhaoyuan; Liu, Yu; Zheng, Rui; Dai, Yao; Tao, Ran; Xia, Huitang; Liu, Hairong; Zhang, Zhiyong; Zhang, Wenjie; Liu, Wei; Cao, Yilin; Zhou, Guangdong

    2016-11-07

    : In vitro three-dimensional (3D) cartilage regeneration is a promising strategy for repair of cartilage defects. However, inferior mechanical strength and tissue homogeneity greatly restricted its clinical translation. Simulation of mechanical stress through a bioreactor is an important approach for improving in vitro cartilage regeneration. The current study developed a hydrostatic pressure (HP) bioreactor based on a novel pressure-transmitting mode achieved by slight deformation of a flexible membrane in a completely sealed stainless steel device. The newly developed bioreactor efficiently avoided the potential risks of previously reported pressure-transmitting modes and simultaneously addressed a series of important issues, such as pressure scopes, culture chamber sizes, sealability, contamination control, and CO2 balance. The whole bioreactor system realized stable long-term (8 weeks) culture under high HP (5-10 MPa) without the problems of medium leakage and contamination. Furthermore, the results of in vitro 3D tissue culture based on a cartilage regeneration model revealed that HP provided by the newly developed bioreactor efficiently promoted in vitro 3D cartilage formation by improving its mechanical strength, thickness, and homogeneity. Detailed analysis in cell proliferation, cartilage matrix production, and cross-linking level of collagen macromolecules, as well as density and alignment of collagen fibers, further revealed the possible mechanisms that HP regulated in vitro cartilage regeneration. The current study provided a highly efficient and stable bioreactor system for improving in vitro 3D cartilage regeneration and thus will help to accelerate its clinical translation. Inferior mechanical strength and tissue homogeneity of in vitro engineered three-dimensional (3D) cartilage greatly restricted its clinical translation. The current study developed a hydrostatic pressure (HP) bioreactor based on a novel pressure-transmitting mode, which

  7. Bioreactor Engineering of Stem Cell Environments

    Science.gov (United States)

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

    2013-01-01

    Stem cells hold promise to revolutionize modern medicine by 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 translation of stem cell based therapies into the clinic. PMID:23531529

  8. Production of manganese peroxidase and laccase in a solid-state bioreactor and modeling of enzyme production kinetics.

    Science.gov (United States)

    Moilanen, Ulla; Winquist, Erika; Mattila, Tuomas; Hatakka, Annele; Eerikäinen, Tero

    2015-01-01

    Lignin-modifying enzymes have various promising applications such as biobleaching, biopulping, the functionalization of lignocellulosic materials, the modification of wood fibers, the remediation of contaminated soil and effluents, as well as improvement of the enzymatic hydrolysis of lignocellulosic substrates. In this study, the production of laccase and manganese peroxidase (MnP) in solid-state cultivation was examined. Oat husks were used as an inexpensive substrate for the white-rot fungus Cerrena unicolor PM170798 (FBCC 387). The addition of a fines fraction (consisting of oat flour and finely ground husks) enhanced MnP production fivefold and laccase production almost threefold. The enzyme production was studied first on a 100 g scale, and the cultivation experiments were then repeated at a larger laboratory-scale (4 kg) in a solid-state bioreactor. High enzyme activity levels were obtained (MnP: 340 nkat g(-1) DM, laccase: 470 nkat g(-1) DM). In addition, the correlation between the CO2 evolution rate and enzyme production was mathematically modeled from the bioreactor experimental data. The model parameters could be used to predict enzyme production.

  9. Biomass viability: An experimental study and the development of an empirical mathematical model for submerged membrane bioreactor.

    Science.gov (United States)

    Zuthi, M F R; Ngo, H H; Guo, W S; Nghiem, L D; Hai, F I; Xia, S Q; Zhang, Z Q; Li, J X

    2015-08-01

    This study investigates the influence of key biomass parameters on specific oxygen uptake rate (SOUR) in a sponge submerged membrane bioreactor (SSMBR) to develop mathematical models of biomass viability. Extra-cellular polymeric substances (EPS) were considered as a lumped parameter of bound EPS (bEPS) and soluble microbial products (SMP). Statistical analyses of experimental results indicate that the bEPS, SMP, mixed liquor suspended solids and volatile suspended solids (MLSS and MLVSS) have functional relationships with SOUR and their relative influence on SOUR was in the order of EPS>bEPS>SMP>MLVSS/MLSS. Based on correlations among biomass parameters and SOUR, two independent empirical models of biomass viability were developed. The models were validated using results of the SSMBR. However, further validation of the models for different operating conditions is suggested.

  10. 浅谈生物反应器在大规模动物细胞培养中的应用%A Brief Discussion on the Application of Bioreactor to Large-Scale Animal Cell Culture

    Institute of Scientific and Technical Information of China (English)

    吴茂柏

    2011-01-01

    生物反应器在大规模动物细胞培养中应用广泛,是大规模细胞培养的关键技术环节。本文对目前主要的一些生物反应器种类、特点和在大规模细胞培养中的应用进行了简要的介绍。%The wide application of bioreactor to large-scale animal cell culture is the key technical link in large-scale animal cell culture.This paper briefly introduces the kinds and features of commonly used bioreactors and their application in large-scale animal cell culture.

  11. 3D Printed Vascular Networks Enhance Viability in High-Volume Perfusion Bioreactor.

    Science.gov (United States)

    Ball, Owen; Nguyen, Bao-Ngoc B; Placone, Jesse K; Fisher, John P

    2016-12-01

    There is a significant clinical need for engineered bone graft substitutes that can quickly, effectively, and safely repair large segmental bone defects. One emerging field of interest involves the growth of engineered bone tissue in vitro within bioreactors, the most promising of which are perfusion bioreactors. Using bioreactor systems, tissue engineered bone constructs can be fabricated in vitro. However, these engineered constructs lack inherent vasculature and once implanted, quickly develop a necrotic core, where no nutrient exchange occurs. Here, we utilized COMSOL modeling to predict oxygen diffusion gradients throughout aggregated alginate constructs, which allowed for the computer-aided design of printable vascular networks, compatible with any large tissue engineered construct cultured in a perfusion bioreactor. We investigated the effect of 3D printed macroscale vascular networks with various porosities on the viability of human mesenchymal stem cells in vitro, using both gas-permeable, and non-gas permeable bioreactor growth chamber walls. Through the use of 3D printed vascular structures in conjunction with a tubular perfusion system bioreactor, cell viability was found to increase by as much as 50% in the core of these constructs, with in silico modeling predicting construct viability at steady state.

  12. Application of high-throughput mini-bioreactor system for systematic scale-down modeling, process characterization, and control strategy development.

    Science.gov (United States)

    Janakiraman, Vijay; Kwiatkowski, Chris; Kshirsagar, Rashmi; Ryll, Thomas; Huang, Yao-Ming

    2015-01-01

    High-throughput systems and processes have typically been targeted for process development and optimization in the bioprocessing industry. For process characterization, bench scale bioreactors have been the system of choice. Due to the need for performing different process conditions for multiple process parameters, the process characterization studies typically span several months and are considered time and resource intensive. In this study, we have shown the application of a high-throughput mini-bioreactor system viz. the Advanced Microscale Bioreactor (ambr15(TM) ), to perform process characterization in less than a month and develop an input control strategy. As a pre-requisite to process characterization, a scale-down model was first developed in the ambr system (15 mL) using statistical multivariate analysis techniques that showed comparability with both manufacturing scale (15,000 L) and bench scale (5 L). Volumetric sparge rates were matched between ambr and manufacturing scale, and the ambr process matched the pCO2 profiles as well as several other process and product quality parameters. The scale-down model was used to perform the process characterization DoE study and product quality results were generated. Upon comparison with DoE data from the bench scale bioreactors, similar effects of process parameters on process yield and product quality were identified between the two systems. We used the ambr data for setting action limits for the critical controlled parameters (CCPs), which were comparable to those from bench scale bioreactor data. In other words, the current work shows that the ambr15(TM) system is capable of replacing the bench scale bioreactor system for routine process development and process characterization.

  13. Spaceflight bioreactor studies of cells and tissues.

    Science.gov (United States)

    Freed, Lisa E; Vunjak-Novakovic, Gordana

    2002-01-01

    Studies of the fundamental role of gravity in the development and function of biological organisms are a central component of the human exploration of space. Microgravity affects numerous physical phenomena relevant to biological research, including the hydrostatic pressure in fluid filled vesicles, sedimentation of organelles, and buoyancy-driven convection of flow and heat. These physical phenomena can in turn directly and indirectly affect cellular morphology, metabolism, locomotion, secretion of extracellular matrix and soluble signals, and assembly into functional tissues. Studies aimed at distinguishing specific effects of gravity on biological systems require the ability to: (i) control and systematically vary gravity, e.g. by utilizing the microgravity environment of space in conjunction with an in-flight centrifuge; and (ii) maintain constant all other factors in the immediate environment, including in particular concentrations and exchange rates of biochemical species and hydrodynamic shear. The latter criteria imply the need for gravity-independent mechanisms to provide for mass transport between the cells and their environment. Available flight hardware has largely determined the experimental design and scientific objectives of spaceflight cell and tissue culture studies carried out to date. Simple culture vessels have yielded important quantitative data, and helped establish in vitro models of cell locomotion, growth and differentiation in various mammalian cell types including embryonic lung cells [6], lymphocytes [2,8], and renal cells [7,31]. Studies done using bacterial cells established the first correlations between gravity-dependent factors such as cell settling velocity and diffusional distance and the respective cell responses [12]. The development of advanced bioreactors for microgravity cell and tissue culture and for tissue engineering has benefited both research areas and provided relevant in vitro model systems for studies of astronaut

  14. Quorum-sensing systems LuxS/autoinducer 2 and Com regulate Streptococcus pneumoniae biofilms in a bioreactor with living cultures of human respiratory cells.

    Science.gov (United States)

    Vidal, Jorge E; Howery, Kristen E; Ludewick, Herbert P; Nava, Porfirio; Klugman, Keith P

    2013-04-01

    Streptococcus pneumoniae forms organized biofilms in the human upper respiratory tract that may play an essential role in both persistence and acute respiratory infection. However, the production and regulation of biofilms on human cells is not yet fully understood. In this work, we developed a bioreactor with living cultures of human respiratory epithelial cells (HREC) and a continuous flow of nutrients, mimicking the microenvironment of the human respiratory epithelium, to study the production and regulation of S. pneumoniae biofilms (SPB). SPB were also produced under static conditions on immobilized HREC. Our experiments demonstrated that the biomass of SPB increased significantly when grown on HREC compared to the amount on abiotic surfaces. Additionally, pneumococcal strains produced more early biofilms on lung cells than on pharyngeal cells. Utilizing the bioreactor or immobilized human cells, the production of early SPB was found to be regulated by two quorum-sensing systems, Com and LuxS/AI-2, since a mutation in either comC or luxS rendered the pneumococcus unable to produce early biofilms on HREC. Interestingly, while LuxS/autoinducer 2 (AI-2) regulated biofilms on both HREC and abiotic surfaces, Com control was specific for those structures produced on HREC. The biofilm phenotypes of strain D39-derivative ΔcomC and ΔluxS QS mutants were reversed by genetic complementation. Of note, SPB formed on immobilized HREC and incubated under static conditions were completely lysed 24 h postinoculation. Biofilm lysis was also regulated by the Com and LuxS/AI-2 quorum-sensing systems.

  15. 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.

  16. Modeling three-dimensional surface morphology of biocake layer in a membrane bioreactor based on fractal geometry.

    Science.gov (United States)

    Zhao, Leihong; Yang, Lining; Lin, Hongjun; Zhang, Meijia; Yu, Haiying; Liao, Bao-Qiang; Wang, Fangyuan; Zhou, Xiaoling; Li, Renjie

    2016-12-01

    While the adsorptive fouling in membrane bioreactors (MBRs) is highly dependent of the surface morphology, little progress has been made on modeling biocake layer surface morphology. In this study, a novel method, which combined static light scattering method for fractal dimension (Df) measurement with fractal method represented by the modified two-variable Weierstrass-Mandelbrot function, was proposed to model biocake layer surface in a MBR. Characterization by atomic force microscopy showed that the biocake surface was stochastic, disorder, self-similarity, and with non-integer dimension, illustrating obvious fractal features. Fractal dimension (Df) of sludge suspension experienced a significant change with operation of the MBR. The constructed biocake layer surface by the proposed method was quite close to the real surface, showing the feasibility of the proposed method. It was found that Df was the critical factor affecting surface morphology, while other factors exerted moderate or minor effects on the roughness of biocake layer.

  17. Performance of a partially packed charcoal pellet bioreactor for acetic acid fermentation.

    Science.gov (United States)

    Horiuchi, J; Ando, K; Watanabe, S; Tada, K; Kobayashi, M; Kanno, T

    2001-01-01

    The performance of a partially packed charcoal pellet bioreactor was compared to that of a fully packed bioreactor for aerobic acetic acid production. In the fully packed charcoal pellet bioreactor, it was considered that the shortening of an actual retention time of the culture broth limited the bioreactor performance under high dilution rate and high aeration conditions. By reducing the filling ratio of charcoal pellets to 44%, which increased the actual retention time of the culture broth, the maximum productivity increased from 3.9 g/l/h in the fully packed bed bioreactor to 5.7 g/l/h in the partially packed bioreactor without affecting the operational stability.

  18. Integrated ‘omics analysis for studying the microbial community response to a pH perturbation of a cellulose-degrading bioreactor culture

    Energy Technology Data Exchange (ETDEWEB)

    Boaro, Amy A.; Kim, Young-Mo; Konopka, Allan; Callister, Stephen J.; Ahring, Birgitte K.

    2014-12-01

    Integrated ‘omics have been used on pure cultures and co-cultures, yet they have not been applied to complex microbial communities to examine questions of perturbation response. In this study, we used integrated ‘omics to measure the perturbation response of a cellulose-degrading bioreactor community fed with microcrystalline cellulose (Avicel). We predicted that a pH decrease by addition of a pulse of acid would reduce microbial community diversity and temporarily reduce reactor function such as cellulose degradation. However, 16S rDNA pyrosequencing results revealed increased alpha diversity in the microbial community after the perturbation, and a persistence of the dominant community members over the duration of the experiment. Proteomics results showed a decrease in activity of proteins associated with Fibrobacter succinogenes two days after the perturbation followed by increased protein abundances six days after the perturbation. The decrease in cellulolytic activity suggested by the proteomics was confirmed by the accumulation of Avicel in the reactor. Metabolomics showed a pattern similar to that of the proteome, with amino acid production decreasing two days after the perturbation and increasing after six days. This study demonstrated that community ‘omics data provides valuable information about the interactions and function of anaerobic cellulolytic community members after a perturbation.

  19. Environmental Control in Flow Bioreactors

    Directory of Open Access Journals (Sweden)

    Serena Giusti

    2017-04-01

    Full Text Available The realization of physiologically-relevant advanced in vitro models is not just related to the reproduction of a three-dimensional multicellular architecture, but also to the maintenance of a cell culture environment in which parameters, such as temperature, pH, and hydrostatic pressure are finely controlled. Tunable and reproducible culture conditions are crucial for the study of environment-sensitive cells, and can also be used for mimicking pathophysiological conditions related with alterations of temperature, pressure and pH. Here, we present the SUITE (Supervising Unit for In Vitro Testing system, a platform able to monitor and adjust local environmental variables in dynamic cell culture experiments. The physical core of the control system is a mixing chamber, which can be connected to different bioreactors and acts as a media reservoir equipped with a pH meter and pressure sensors. The chamber is heated by external resistive elements and the temperature is controlled using a thermistor. A purpose-built electronic control unit gathers all data from the sensors and controls the pH and hydrostatic pressure by regulating air and CO2 overpressure and flux. The system’s modularity and the possibility of imposing different pressure conditions were used to implement a model of portal hypertension with both endothelial and hepatic cells. The results show that the SUITE platform is able to control and maintain cell culture parameters at fixed values that represent either physiological or pathological conditions. Thus, it represents a fundamental tool for the design of biomimetic in vitro models, with applications in disease modelling or toxicity testing.

  20. Modeling and CFD simulation of nutrient distribution in picoliter bioreactors for bacterial growth studies on single-cell level.

    Science.gov (United States)

    Westerwalbesloh, Christoph; Grünberger, Alexander; Stute, Birgit; Weber, Sophie; Wiechert, Wolfgang; Kohlheyer, Dietrich; von Lieres, Eric

    2015-11-01

    A microfluidic device for microbial single-cell cultivation of bacteria was modeled and simulated using COMSOL Multiphysics. The liquid velocity field and the mass transfer within the supply channels and cultivation chambers were calculated to gain insight in the distribution of supplied nutrients and metabolic products secreted by the cultivated bacteria. The goal was to identify potential substrate limitations or product accumulations within the cultivation device. The metabolic uptake and production rates, colony size, and growth medium composition were varied covering a wide range of operating conditions. Simulations with glucose as substrate did not show limitations within the typically used concentration range, but for alternative substrates limitations could not be ruled out. This lays the foundation for further studies and the optimization of existing picoliter bioreactor systems.

  1. Cultural models of linguistic standardization

    Directory of Open Access Journals (Sweden)

    Dirk Geeraerts

    2016-02-01

    Full Text Available In line with well-known trends in cultural theory (see Burke et al., 2000, Cognitive Linguistics has stressed the idea that we think about social reality in terms of models – ‘cultural models’ or ‘folk theories’: from Holland & Quinn (1987 over Lakoff (1996 and Palmer (1996 to Dirven et al. (2001a, 2001b, Cognitive linguists have demonstrated how the technical apparatus of Cognitive Linguistics can be used to analyze how our conception of social reality is shaped by underlying patterns of thought. But if language is a social and cultural reality, what are the models that shape our conception of language? Specifically, what are the models that shape our thinking about language as a social phenomenon? What are the paradigms that we use to think about language, not primarily in terms of linguistic structure (as in Reddy 1979, but in terms of linguistic variation: models about the way in which language varieties are distributed over a language community and about the way in which such distribution should be evaluated?In this paper, I will argue that two basic models may be identified: a rationalist and a romantic one. I will chart the ways in which they interact, describe how they are transformed in the course of time, and explore how the models can be used in the analysis of actual linguistic variation.

  2. Model-based analysis of the effect of different operating conditions on fouling mechanisms in a membrane bioreactor.

    Science.gov (United States)

    Sabia, Gianpaolo; Ferraris, Marco; Spagni, Alessandro

    2016-01-01

    This study proposes a model-based evaluation of the effect of different operating conditions with and without pre-denitrification treatment and applying three different solids retention times on the fouling mechanisms involved in membrane bioreactors (MBRs). A total of 11 fouling models obtained from literature were used to fit the transmembrane pressure variations measured in a pilot-scale MBR treating real wastewater for more than 1 year. The results showed that all the models represent reasonable descriptions of the fouling processes in the MBR tested. The model-based analysis confirmed that membrane fouling started by pore blocking (complete blocking model) and by a reduction of the pore diameter (standard blocking) while cake filtration became the dominant fouling mechanism over long-term operation. However, the different fouling mechanisms occurred almost simultaneously making it rather difficult to identify each one. The membrane "history" (i.e. age, lifespan, etc.) seems the most important factor affecting the fouling mechanism more than the applied operating conditions. Nonlinear regression of the most complex models (combined models) evaluated in this study sometimes demonstrated unreliable parameter estimates suggesting that the four basic fouling models (complete, standard, intermediate blocking and cake filtration) contain enough details to represent a reasonable description of the main fouling processes occurring in MBRs.

  3. A multiphase model for chemically- and mechanically- induced cell differentiation in a hollow fibre membrane bioreactor: minimising growth factor consumption.

    Science.gov (United States)

    Pearson, Natalie C; Oliver, James M; Shipley, Rebecca J; Waters, Sarah L

    2016-06-01

    We present a simplified two-dimensional model of fluid flow, solute transport, and cell distribution in a hollow fibre membrane bioreactor. We consider two cell populations, one undifferentiated and one differentiated, with differentiation stimulated either by growth factor alone, or by both growth factor and fluid shear stress. Two experimental configurations are considered, a 3-layer model in which the cells are seeded in a scaffold throughout the extracapillary space (ECS), and a 4-layer model in which the cell-scaffold construct occupies a layer surrounding the outside of the hollow fibre, only partially filling the ECS. Above this is a region of free-flowing fluid, referred to as the upper fluid layer. Following previous models by the authors (Pearson et al. in Math Med Biol, 2013, Biomech Model Mechanbiol 1-16, 2014a, we employ porous mixture theory to model the dynamics of, and interactions between, the cells, scaffold, and fluid in the cell-scaffold construct. We use this model to determine operating conditions (experiment end time, growth factor inlet concentration, and inlet fluid fluxes) which result in a required percentage of differentiated cells, as well as maximising the differentiated cell yield and minimising the consumption of expensive growth factor.

  4. Evaluation of biological nutrient removal from wastewater by Twin Circulating Fluidized Bed Bioreactor (TCFBBR) using a predictive fluidization model and AQUIFAS APP.

    Science.gov (United States)

    Andalib, Mehran; Nakhla, George; Sen, Dipankar; Zhu, Jesse

    2011-02-01

    A two-phase and three-phase predictive fluidization model based on the characteristics of a system such as media type and size, flow rates, and reactor cross sectional area was proposed to calculate bed expansion, solid, liquid and gas hold up and specific surface area (SSA) of the biofilm particles. The model was subsequently linked to 1d AQUIFAS APP software (Aquaregen) to model biological nutrient removal in two phase (anoxic) and three phase (aerobic) fluidized bed bioreactors. The credibility of the proposed model for biological nutrient removal was investigated using the experimental data from a Twin Circulating Fluidized Bed Bioreactors (TCFBBR) treating synthetic and municipal wastewater. The SSA of bio-particles and volume of the expanded bed were simulated as a function of operational parameters. Two-sided t-tests demonstrated that simulated SCOD, NH(4)-N, NO(3)-N, TN, VSS and biomass yields agreed with the experimental values at the 95% confidence level.

  5. The evolution of simulation techniques for dynamic bone tissue engineering in bioreactors.

    Science.gov (United States)

    Vetsch, Jolanda Rita; Müller, Ralph; Hofmann, Sandra

    2015-08-01

    Bone tissue engineering aims to overcome the drawbacks of current bone regeneration techniques in orthopaedics. Bioreactors are widely used in the field of bone tissue engineering, as they help support efficient nutrition of cultured cells with the possible combination of applying mechanical stimuli. Beneficial influencing parameters of in vitro cultures are difficult to find and are mostly determined by trial and error, which is associated with significant time and money spent. Mathematical simulations can support the finding of optimal parameters. Simulations have evolved over the last 20 years from simple analytical models to complex and detailed computational models. They allow researchers to simulate the mechanical as well as the biological environment experienced by cells seeded on scaffolds in a bioreactor. Based on the simulation results, it is possible to give recommendations about specific parameters for bone bioreactor cultures, such as scaffold geometries, scaffold mechanical properties, the level of applied mechanical loading or nutrient concentrations. This article reviews the evolution in simulating various aspects of dynamic bone culture in bioreactors and reveals future research directions.

  6. Integrated Safety Culture Model and Application

    Institute of Scientific and Technical Information of China (English)

    汪磊; 孙瑞山; 刘汉辉

    2009-01-01

    A new safety culture model is constructed and is applied to analyze the correlations between safety culture and SMS. On the basis of previous typical definitions, models and theories of safety culture, an in-depth analysis on safety culture's structure, composing elements and their correlations was conducted. A new definition of safety culture was proposed from the perspective of sub-cuhure. 7 types of safety sub-culture, which are safety priority culture, standardizing culture, flexible culture, learning culture, teamwork culture, reporting culture and justice culture were defined later. Then integrated safety culture model (ISCM) was put forward based on the definition. The model divided safety culture into intrinsic latency level and extrinsic indication level and explained the potential relationship between safety sub-culture and all safety culture dimensions. Finally in the analyzing of safety culture and SMS, it concluded that positive safety culture is the basis of im-plementing SMS effectively and an advanced SMS will improve safety culture from all around.

  7. Induction of Hepatic and Endothelial Differentiation by Perfusion in a Three-Dimensional Cell Culture Model of Human Fetal Liver.

    Science.gov (United States)

    Pekor, Christopher; Gerlach, Jörg C; Nettleship, Ian; Schmelzer, Eva

    2015-07-01

    The development of functional engineered tissue constructs depends on high cell densities and appropriate vascularization. In this study we implemented a four-compartment three-dimensional perfusion bioreactor culture model for studying the effects of medium perfusion on endothelial, hepatic, and hematopoietic cell populations of primary human fetal liver in an in vivo-like environment. Human fetal liver cells were cultured in bioreactors configured to provide either perfusion or diffusion conditions. Metabolic activities of the cultures were monitored daily by measuring glucose consumption and lactate production. Cell viability during culture was analyzed by lactate dehydrogenase activity. Hepatic functionality was determined by the release of albumin and alpha-fetoprotein (AFP) in culture medium samples. After 4 days of culture, cells were analyzed for the expression of a variety of endothelial, hepatic, and hematopoietic genes, as well as the surface marker expression of CD31 and CD34 in flow cytometry. We found that medium perfusion increased the gene expression of endothelial markers such as CD31, von Willebrand factor (vWF), CD140b, CD309, and CD144 while decreasing the gene expression of the erythrocyte-surface marker CD235a. Hepatic differentiation was promoted under perfusion conditions as demonstrated by lower AFP and higher albumin secretion compared with cultures not exposed to medium perfusion. Additionally, cultures exposed to medium perfusion gave higher rates of glucose consumption and lactate production, indicating increased metabolic activity. In conclusion, high-density bioreactors configured to provide constant medium perfusion significantly induced hepatic and endothelial cell differentiation and provided improved conditions for the culture of human fetal liver cells compared with cultures without perfusion.

  8. Monolithic Continuous-Flow Bioreactors

    Science.gov (United States)

    Stephanopoulos, Gregory; Kornfield, Julia A.; Voecks, Gerald A.

    1993-01-01

    Monolithic ceramic matrices containing many small flow passages useful as continuous-flow bioreactors. Ceramic matrix containing passages made by extruding and firing suitable ceramic. Pores in matrix provide attachment medium for film of cells and allow free movement of solution. Material one not toxic to micro-organisms grown in reactor. In reactor, liquid nutrients flow over, and liquid reaction products flow from, cell culture immobilized in one set of channels while oxygen flows to, and gaseous reaction products flow from, culture in adjacent set of passages. Cells live on inner surfaces containing flowing nutrient and in pores of walls of passages. Ready access to nutrients and oxygen in channels. They generate continuous high yield characteristic of immobilized cells, without large expenditure of energy otherwise incurred if necessary to pump nutrient solution through dense biomass as in bioreactors of other types.

  9. Region specific response of intervertebral disc cells to complex dynamic loading: an organ culture study using a dynamic torsion-compression bioreactor.

    Directory of Open Access Journals (Sweden)

    Samantha C W Chan

    Full Text Available The spine is routinely subjected to repetitive complex loading consisting of axial compression, torsion, flexion and extension. Mechanical loading is one of the important causes of spinal diseases, including disc herniation and disc degeneration. It is known that static and dynamic compression can lead to progressive disc degeneration, but little is known about the mechanobiology of the disc subjected to combined dynamic compression and torsion. Therefore, the purpose of this study was to compare the mechanobiology of the intervertebral disc when subjected to combined dynamic compression and axial torsion or pure dynamic compression or axial torsion using organ culture. We applied four different loading modalities [1. control: no loading (NL, 2. cyclic compression (CC, 3. cyclic torsion (CT, and 4. combined cyclic compression and torsion (CCT] on bovine caudal disc explants using our custom made dynamic loading bioreactor for disc organ culture. Loads were applied for 8 h/day and continued for 14 days, all at a physiological magnitude and frequency. Our results provided strong evidence that complex loading induced a stronger degree of disc degeneration compared to one degree of freedom loading. In the CCT group, less than 10% nucleus pulposus (NP cells survived the 14 days of loading, while cell viabilities were maintained above 70% in the NP of all the other three groups and in the annulus fibrosus (AF of all the groups. Gene expression analysis revealed a strong up-regulation in matrix genes and matrix remodeling genes in the AF of the CCT group. Cell apoptotic activity and glycosaminoglycan content were also quantified but there were no statistically significant differences found. Cell morphology in the NP of the CCT was changed, as shown by histological evaluation. Our results stress the importance of complex loading on the initiation and progression of disc degeneration.

  10. Developing a scalable model of recombinant protein yield from Pichia pastoris: the influence of culture conditions, biomass and induction regime

    Directory of Open Access Journals (Sweden)

    Wilks Martin DB

    2009-07-01

    Full Text Available Abstract Background The optimisation and scale-up of process conditions leading to high yields of recombinant proteins is an enduring bottleneck in the post-genomic sciences. Typical experiments rely on varying selected parameters through repeated rounds of trial-and-error optimisation. To rationalise this, several groups have recently adopted the 'design of experiments' (DoE approach frequently used in industry. Studies have focused on parameters such as medium composition, nutrient feed rates and induction of expression in shake flasks or bioreactors, as well as oxygen transfer rates in micro-well plates. In this study we wanted to generate a predictive model that described small-scale screens and to test its scalability to bioreactors. Results Here we demonstrate how the use of a DoE approach in a multi-well mini-bioreactor permitted the rapid establishment of high yielding production phase conditions that could be transferred to a 7 L bioreactor. Using green fluorescent protein secreted from Pichia pastoris, we derived a predictive model of protein yield as a function of the three most commonly-varied process parameters: temperature, pH and the percentage of dissolved oxygen in the culture medium. Importantly, when yield was normalised to culture volume and density, the model was scalable from mL to L working volumes. By increasing pre-induction biomass accumulation, model-predicted yields were further improved. Yield improvement was most significant, however, on varying the fed-batch induction regime to minimise methanol accumulation so that the productivity of the culture increased throughout the whole induction period. These findings suggest the importance of matching the rate of protein production with the host metabolism. Conclusion We demonstrate how a rational, stepwise approach to recombinant protein production screens can reduce process development time.

  11. The modeling of ethanol production by Kluyveromyces marxianus using whey as substrate in continuous A-Stat bioreactors.

    Science.gov (United States)

    Gabardo, Sabrina; Pereira, Gabriela Feix; Rech, Rosane; Ayub, Marco Antônio Záchia

    2015-09-01

    We investigated the kinetics of whey bioconversion into ethanol by Kluyveromyces marxianus in continuous bioreactors using the "accelerostat technique" (A-stat). Cultivations using free and Ca-alginate immobilized cells were evaluated using two different acceleration rates (a). The kinetic profiles of these systems were modeled using four different unstructured models, differing in the expressions for the specific growth (μ) and substrate consumption rates (r s), taking into account substrate limitation and product inhibition. Experimental data showed that the dilution rate (D) directly affected cell physiology and metabolism. The specific growth rate followed the dilution rate (μ≈D) for the lowest acceleration rate (a = 0.0015 h(-2)), condition in which the highest ethanol yield (0.52 g g(-1)) was obtained. The highest acceleration rate (a = 0.00667 h(-2)) led to a lower ethanol yield (0.40 g g(-1)) in the system where free cells were used, whereas with immobilized cells ethanol yields increased by 23 % (0.49 g g(-1)). Among the evaluated models, Monod and Levenspiel combined with Ghose and Tyagi models were found to be more appropriate for describing the kinetics of whey bioconversion into ethanol. These results may be useful in scaling up the process for ethanol production from whey.

  12. 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

    2013-01-01

    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 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. PMID:23072472

  13. An automated perfusion bioreactor for the streamlined production of engineered osteogenic grafts.

    Science.gov (United States)

    Ding, Ming; Henriksen, Susan S; Wendt, David; Overgaard, Søren

    2016-04-01

    A computer-controlled perfusion bioreactor was developed for the streamlined production of engineered osteogenic grafts. This system automated the required bioprocesses, from the initial filling of the system through the phases of cell seeding and prolonged cell/tissue culture. Flow through chemo-optic micro-sensors allowed to non-invasively monitor the levels of oxygen and pH in the perfused culture medium throughout the culture period. To validate its performance, freshly isolated ovine bone marrow stromal cells were directly seeded on porous scaffold granules (hydroxyapatite/β-tricalcium-phosphate/poly-lactic acid), bypassing the phase of monolayer cell expansion in flasks. Either 10 or 20 days after culture, engineered cell-granule grafts were implanted in an ectopic mouse model to quantify new bone formation. After four weeks of implantation, histomorphometry showed more bone in bioreactor-generated grafts than cell-free granule controls, while bone formation did not show significant differences between 10 days and 20 days of incubation. The implanted granules without cells had no bone formation. This novel perfusion bioreactor has revealed the capability of activation larger viable bone graft material, even after shorter incubation time of graft material. This study has demonstrated the feasibility of engineering osteogenic grafts in an automated bioreactor system, laying the foundation for a safe, regulatory-compliant, and cost-effective manufacturing process. © 2015 Wiley Periodicals, Inc.

  14. Three-Dimensional Rotating Wall Vessel-Derived Cell Culture Models for Studying Virus-Host Interactions

    Directory of Open Access Journals (Sweden)

    Jameson K. Gardner

    2016-11-01

    Full Text Available The key to better understanding complex virus-host interactions is the utilization of robust three-dimensional (3D human cell cultures that effectively recapitulate native tissue architecture and model the microenvironment. A lack of physiologically-relevant animal models for many viruses has limited the elucidation of factors that influence viral pathogenesis and of complex host immune mechanisms. Conventional monolayer cell cultures may support viral infection, but are unable to form the tissue structures and complex microenvironments that mimic host physiology and, therefore, limiting their translational utility. The rotating wall vessel (RWV bioreactor was designed by the National Aeronautics and Space Administration (NASA to model microgravity and was later found to more accurately reproduce features of human tissue in vivo. Cells grown in RWV bioreactors develop in a low fluid-shear environment, which enables cells to form complex 3D tissue-like aggregates. A wide variety of human tissues (from neuronal to vaginal tissue have been grown in RWV bioreactors and have been shown to support productive viral infection and physiological meaningful host responses. The in vivo-like characteristics and cellular features of the human 3D RWV-derived aggregates make them ideal model systems to effectively recapitulate pathophysiology and host responses necessary to conduct rigorous basic science, preclinical and translational studies.

  15. Controlled-Turbulence Bioreactors

    Science.gov (United States)

    Wolf, David A.; Schwartz, Ray; Trinh, Tinh

    1989-01-01

    Two versions of bioreactor vessel provide steady supplies of oxygen and nutrients with little turbulence. Suspends cells in environment needed for sustenance and growth, while inflicting less damage from agitation and bubbling than do propeller-stirred reactors. Gentle environments in new reactors well suited to delicate mammalian cells. One reactor kept human kidney cells alive for as long as 11 days. Cells grow on carrier beads suspended in liquid culture medium that fills cylindrical housing. Rotating vanes - inside vessel but outside filter - gently circulates nutrient medium. Vessel stationary; magnetic clutch drives filter cylinder and vanes. Another reactor creates even less turbulence. Oxygen-permeable tubing wrapped around rod extending along central axis. Small external pump feeds oxygen to tubing through rotary coupling, and oxygen diffuses into liquid medium.

  16. A systematic approach for scale-down model development and characterization of commercial cell culture processes.

    Science.gov (United States)

    Li, Feng; Hashimura, Yasunori; Pendleton, Robert; Harms, Jean; Collins, Erin; Lee, Brian

    2006-01-01

    The objective of process characterization is to demonstrate robustness of manufacturing processes by understanding the relationship between key operating parameters and final performance. Technical information from the characterization study is important for subsequent process validation, and this has become a regulatory expectation in recent years. Since performing the study at the manufacturing scale is not practically feasible, development of scale-down models that represent the performance of the commercial process is essential to achieve reliable process characterization. In this study, we describe a systematic approach to develop a bioreactor scale-down model and to characterize a cell culture process for recombinant protein production in CHO cells. First, a scale-down model using 2-L bioreactors was developed on the basis of the 2000-L commercial scale process. Profiles of cell growth, productivity, product quality, culture environments (pH, DO, pCO2), and level of metabolites (glucose, glutamine, lactate, ammonia) were compared between the two scales to qualify the scale-down model. The key operating parameters were then characterized in single-parameter ranging studies and an interaction study using this scale-down model. Appropriate operation ranges and acceptance criteria for certain key parameters were determined to ensure the success of process validation and the process performance consistency. The process worst-case condition was also identified through the interaction study.

  17. 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.

  18. Modelling the removal of volatile pollutants under transient conditions in a two-stage bioreactor using artificial neural networks.

    Science.gov (United States)

    López, M Estefanía; Rene, Eldon R; Boger, Zvi; Veiga, María C; Kennes, Christian

    2017-02-15

    A two-stage biological waste gas treatment system consisting of a first stage biotrickling filter (BTF) and second stage biofilter (BF) was tested for the removal of a gas-phase methanol (M), hydrogen sulphide (HS) and α-pinene (P) mixture. The bioreactors were tested with two types of shock loads, i.e., long-term (66h) low to medium concentration loads, and short-term (12h) low to high concentration loads. M and HS were removed in the BTF, reaching maximum elimination capacities (ECmax) of 684 and 33 gm(-3)h(-1), respectively. P was removed better in the second stage BF with an ECmax of 130 gm(-3)h(-1). The performance was modelled using two multi-layer perceptrons (MLPs) that employed the error backpropagation with momentum algorithm, in order to predict the removal efficiencies (RE, %) of methanol (REM), hydrogen sulphide (REHS) and α-pinene (REP), respectively. It was observed that, a MLP with the topology 3-4-2 was able to predict REM and REHS in the BTF, while a topology of 3-3-1 was able to approximate REP in the BF. The results show that artificial neural network (ANN) based models can effectively be used to model the transient-state performance of bioprocesses treating gas-phase pollutants.

  19. Advanced Wastewater Treatment Engineering—Investigating Membrane Fouling in both Rotational and Static Membrane Bioreactor Systems Using Empirical Modelling

    Directory of Open Access Journals (Sweden)

    Parneet Paul

    2016-01-01

    Full Text Available Advanced wastewater treatment using membranes are popular environmental system processes since they allow reuse and recycling. However, fouling is a key limiting factor and so proprietary systems such as Avanti’s RPU-185 Flexidisks membrane bioreactor (MBR use novel rotating membranes to assist in ameliorating it. In earlier research, this rotating process was studied by creating a simulation model based on first principles and traditional fouling mechanisms. In order to directly compare the potential benefits of this rotational system, this follow-up study was carried out using Avanti’s newly developed static (non-rotating Flexidisks MBR system. The results from operating the static pilot unit were simulated and modelled using the rotational fouling model developed earlier however with rotational switching functions turned off and rotational parameters set to a static mode. The study concluded that a rotating MBR system could increase flux throughput when compared against a similar static system. It is thought that although the slowly rotating spindle induces a weak crossflow shear, it is still able to even out cake build up across the membrane surface, thus reducing the likelihood of localised critical flux being exceeded at the micro level and lessening the potential of rapid trans-membrane pressure increases at the macro level.

  20. Advanced Wastewater Treatment Engineering-Investigating Membrane Fouling in both Rotational and Static Membrane Bioreactor Systems Using Empirical Modelling.

    Science.gov (United States)

    Paul, Parneet; Jones, Franck Anderson

    2016-01-05

    Advanced wastewater treatment using membranes are popular environmental system processes since they allow reuse and recycling. However, fouling is a key limiting factor and so proprietary systems such as Avanti's RPU-185 Flexidisks membrane bioreactor (MBR) use novel rotating membranes to assist in ameliorating it. In earlier research, this rotating process was studied by creating a simulation model based on first principles and traditional fouling mechanisms. In order to directly compare the potential benefits of this rotational system, this follow-up study was carried out using Avanti's newly developed static (non-rotating) Flexidisks MBR system. The results from operating the static pilot unit were simulated and modelled using the rotational fouling model developed earlier however with rotational switching functions turned off and rotational parameters set to a static mode. The study concluded that a rotating MBR system could increase flux throughput when compared against a similar static system. It is thought that although the slowly rotating spindle induces a weak crossflow shear, it is still able to even out cake build up across the membrane surface, thus reducing the likelihood of localised critical flux being exceeded at the micro level and lessening the potential of rapid trans-membrane pressure increases at the macro level.

  1. Slurry bioreactor modeling using a dissimilatory arsenate-reducing bacterium for remediation of arsenic-contaminated soil.

    Science.gov (United States)

    Soda, Satoshi; Kanzaki, Masaya; Yamamuara, Shigeki; Kashiwa, Masami; Fujita, Masanori; Ike, Michihiko

    2009-02-01

    A slurry bioreactor using a dissimilatory arsenate (As(V))-reducing bacterium is proposed for remediation of arsenic-contaminated soils. Bacterial As(V) reduction can cause arsenic extraction from the solid to the liquid phase because arsenite, As(III), is much less adsorptive than As(V). A mathematical model was developed incorporating the reversible sorption process of arsenic as well as bacterial growth and decay via As(V) reduction. A linear isotherm equation expressed the sorption process. The model included Haldane kinetics with high As(V) concentrations and cell inactivation by toxicity due to As(III). Extraction experiments used synthetic contaminated soils (forest soil, Soil SF, 1100 mg kg(-1); paddy soil, Soil SP, 1100 mg kg(-1)) and actual contaminated soils (Soil AH 2200 mg kg(-1) and Soil AL, 220 mg kg(-1)) at 5% w/v slurry concentration. Simulation results matched the observed changes of arsenic concentrations in the liquid phase. The respective extraction efficiencies of arsenic were 63%, 41%, 20%, and 55% for SF, SP, AH, and AL soils. Sensitivity analyses showed that the rate-limiting step was the desorption rate of As(V) from the solid to the liquid phase, rather than the As(V)-reducing rate. The proposed model provides a useful framework for understanding and predicting the extraction of arsenic from soil.

  2. Reduced-Gravity Experiments Conducted to Help Bioreactor Development

    Science.gov (United States)

    Niederhaus, Charles E.; Nahra, Henry K.; Kizito, John P.

    2004-01-01

    The NASA Glenn Research Center and the NASA Johnson Space Center are collaborating on fluid dynamic investigations for a future cell science bioreactor to fly on the International Space Station (ISS). Project Manager Steven Gonda from the Cellular Biotechnology Program at Johnson is leading the development of the Hydrodynamic Focusing Bioreactor--Space (HFB-S) for use on the ISS to study tissue growth in microgravity. Glenn is providing microgravity fluid physics expertise to help with the design and evaluation of the HFB-S. These bioreactors are used for three-dimensional tissue culture, which cannot be done in ground-based labs in normal gravity. The bioreactors provide a continual supply of oxygen for cell growth, as well as periodic replacement of cell culture media with nutrients. The bioreactor must provide a uniform distribution of oxygen and nutrients while minimizing the shear stresses on the tissue culture.

  3. Optimal homogenization of perfusion flows in microfluidic bio-reactors; a numerical study

    CERN Document Server

    Okkels, Fridolin; Bruus, Henrik

    2009-01-01

    To ensure homogeneous conditions within the complete area of perfused microfluidic bio-reactors, we develop a general design of a continuously feed bio-reactor with uniform perfusion flow. This is achieved by introducing a specific type of perfusion inlet to the reaction area. The geometry of these inlets are found using the methods of topology optimization and shape optimization. The results are compared with two different analytic models, from which a general parametric description of the design is obtained and tested numerically. Such a parametric description will generally be beneficial for the design of a broad range of microfluidic bioreactors used for e.g. cell culturing and analysis, and in feeding bio-arrays.

  4. Cell Separations in Microgravity and Development of a Space Bioreactor

    Science.gov (United States)

    Morrison, D. R.

    1985-01-01

    A bioreactor optimized for operations in space is now being developed. The current research is focused on determining the optimum cell-bead ratios, medium content and proper maintenance conditions required to keep living cell specimens alive and healthy for the entire flight. The bioreactor development project has recently added a microprocessor/computer to the JSC prototype for control and data analysis. Appropriate new technology is being combined with the current bioreactor designs and tested to determine what specific features must be included in the fabrication of a bioreactor designed to operate for STS demonstration tests. Considerations include: (1) circulation and resupply of culture media; (2) sensors required to monitor temperature, cell growth, mass transport, and oxygen consumption; and (3) inflight control of shear stress on cells, gas transfer in microgravity, diffusion, and intracellular transport. These data and results from the JSC prototype bioreactor test will be used for the design and construction of a small space bioreactor for the Orbiter middeck.

  5. Periodic harvesting of embryonic stem cells from a hollow-fiber membrane based four-compartment bioreactor.

    Science.gov (United States)

    Knöspel, Fanny; Freyer, Nora; Stecklum, Maria; Gerlach, Jörg C; Zeilinger, Katrin

    2016-01-01

    Different types of stem cells have been investigated for applications in drug screening and toxicity testing. In order to provide sufficient numbers of cells for such in vitro applications a scale-up of stem cell culture is necessary. Bioreactors for dynamic three-dimensional (3D) culture of growing cells offer the option for culturing large amounts of stem cells at high densities in a closed system. We describe a method for periodic harvesting of pluripotent stem cells (PSC) during expansion in a perfused 3D hollow-fiber membrane bioreactor, using mouse embryonic stem cells (mESC) as a model cell line. A number of 100 × 10(6) mESC were seeded in bioreactors in the presence of mouse embryonic fibroblasts (MEF) as feeder cells. Over a cultivation interval of nine days cells were harvested by trypsin perfusion and mechanical agitation every second to third culture day. A mean of 380 × 10(6) mESC could be removed with every harvest. Subsequent to harvesting, cells continued growing in the bioreactor, as determined by increasing glucose consumption and lactate production. Immunocytochemical staining and mRNA expression analysis of markers for pluripotency and the three germ layers showed a similar expression of most markers in the harvested cells and in mESC control cultures. In conclusion, successful expansion and harvesting of viable mESC from bioreactor cultures with preservation of sterility was shown. The present study is the first one showing the feasibility of periodic harvesting of adherent cells from a continuously perfused four-compartment bioreactor including further cultivation of remaining cells. © 2015 American Institute of Chemical Engineers.

  6. Neural Network Modeling and Prediction of Methane Fraction in Biogas from Landfill Bioreactors

    Directory of Open Access Journals (Sweden)

    A Ghavidel

    2009-09-01

    Full Text Available "n "nBackgrounds and Objectives:A number of different technologies have recently been studied todetermine the best use of biogas, however, to choose optimize technologies of using biogas for energy recovery it is necessary to monitor and predict the methane percentage of biogas. In this study, a method is proposed for predicting the methane fraction in landfill gas originating from Labscalelandfill bioreactors, based on neural network."nMaterials and Methods: In this study, two different systems were applied, to predict the methane fraction in landfill gas as a final product of anaerobic digestion, we used the leachate specifications as input parameters. In system I (C1, the leachate generated from a fresh-waste reactor was drained to recirculation tank, and recycled. In System II (C2, the leachate generated from a fresh waste landfill reactor was fed through a well-decomposed refuse landfill reactor, and at the same time, the leachate generated from a well-decomposed refuse landfill reactor recycled to a fresh waste landfill reactor."nResults: There is very good agreement in the trends between forecasted and measured data. R valuesare 0.999 and 0.997, and the obtained Root mean square error values are 1.098 and 2.387 for training and test data, respectively"nConclusion: The proposed method can significantly predict the methane fraction in landfill gasoriginating and, consequently, neural network can be use to optimize the dimensions of a plant using biogas for energy (i.e. heat and/or electricity recovery and monitoring system.

  7. Virus rejection with two model human enteric viruses in membrane bioreactor system

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    A membrane bioreactor (MBR) with gravity drain was tested for virus rejection with two coliphages, T4 and f2, which were used as surrogates for human enteric viruses. Virus rejection was investigated by PVDF and PP membrane modules, with the pore sizes of 0.22 and 0.1 μm, respectively. In tap water system, 2.1 lg rejection of coliphage T4 could be achieved by PVDF membrane compared with complete rejection by PP membrane, while for coliphage f2 with smaller diameter, 0.3―0.5 lg rejection of the influent virus was removed by the two membranes. In domestic wastewater system, cake layer and gel layer on the membrane surface changed the cut-off size of the membrane so that there was no significant difference between PP and PVDF for each coliphage. The removal ratios of coliphage T4 and f2 in the MBR were more than 5.5 and 3.0 lg, respectively. Compared with 5.5 lg removal for virus T4 in the MBR system, only 2.1 lg (96.8%―99.9%) removal rate was observed in the conventional activated sludge system with the influent virus concentration fluctuating from 1830 to 57000 PFU/mL. Only 0.8%―22% virus removal was the effect of adsorption to activated sludge, which showed a decreasing tendency with the retention time, while 75%―98% was the effect of virus inactivation by microbial activity. It indicated that the major mechanism of virus removal was not the transfer of viruses from the water phase to the sludge phase but inactivation in the biological treatment process.

  8. Virus rejection with two model human enteric viruses in membrane bioreactor system

    Institute of Scientific and Technical Information of China (English)

    ZHENG Xiang; LIU JunXin

    2007-01-01

    A membrane bioreactor (MBR) with gravity drain was tested for virus rejection with two coliphages, T4 and f2, which were used as surrogates for human enteric viruses. Virus rejection was investigated by PVDF and PP membrane modules, with the pore sizes of 0.22 and 0.1 μm, respectively. In tap water system, 2.1 lg rejection of coliphage T4 could be achieved by PVDF membrane compared with complete rejection by PP membrane, while for coliphage f2 with smaller diameter, 0.3-0.5 lg rejection of the influent virus was removed by the two membranes. In domestic wastewater system, cake layer and gel layer on the membrane surface changed the cut-off size of the membrane so that there was no significant difference between PP and PVDF for each coliphage. The removal ratios of coliphage T4 and f2 in the MBR were more than 5.5 and 3.0 lg, respectively. Compared with 5.5 lg removal for virus T4 in the MBR system, only 2.1 lg (96.8%-99.9%) removal rate was observed in the conventional activated sludge system with the influent virus concentration fluctuating from 1830 to 57000 PFU/mL. Only 0.8 %-22 % virus removal was the effect of adsorption to activated sludge, which showed a decreasing tendency with the retention time, while 75%-98% was the effect of virus inactivation by microbial activity. It indicated that the major mechanism of virus removal was not the transfer of viruses from the water phase to the sludge phase but inactivation in the biological treatment process.

  9. Modeling of Oxygen Transfer Correlations for Stirred Tank Bioreactor Agitated with Atypical Helical Ribbon Impeller

    Directory of Open Access Journals (Sweden)

    Mohd S. Mohamed

    2009-01-01

    Full Text Available Problem Statement: Mixing filamentous fungi entails delicate balance between promoting high volumetric oxygen transfer coefficient (kLa while keeping low hydrodynamic stresses in the microenvironment of cultures. This study examined the oxygen transfer capability of a prototype low shear helical impeller as a potential replacement for the standard Rushton turbine. Approach: The dependence of kLa upon specific power uptake, superficial gas velocity and apparent viscosity were examined under coalescent, non-coalescent and viscous pseudoplastic fluids scenario using dynamic gassing out technique. Subsequently, collected data were treated under historical data design of response surface methodology. Results: Three empirical power law kLa correlations were developed for each fluid. Correlation comparison with literature models for single turbine agitation suggest higher kLa augmentation within 1.5-3.6 folds for helical impeller in distilled water and as high as 78% improvement in electrolytes fluid. However impeller performance is comparatively 10% lower against theoretical kLa of triple turbines arrangement for power uptake between 0-1600 W m-3. In case of carboxymethylcellulose, better oxygen transfer is expected for design with higher proportionality of impeller-to-tank internal diameter. Conclusions: Helical impeller performance as gas-liquid contactor is comparable to single and triple Rushtons turbines, with superficial gas velocity proved to be more influential than power uptake in non-viscous liquids and vice versa in viscous pseudoplastic fluids.

  10. Sub-critical fouling in a membrane bioreactor for municipal wastewater treatment: experimental investigation and mathematical modelling.

    Science.gov (United States)

    Guglielmi, Giuseppe; Saroj, Devendra Prakash; Chiarani, Daniele; Andreottola, Gianni

    2007-09-01

    Fouling is a major limitation for the application of membrane bioreactors (MBRs) in municipal wastewater treatment; the critical flux concept represents a valid tool for process optimisation in planning fouling control strategies. The paper presents the results obtained on a large pilot MBR equipped with a plate-and-frame ultrafiltration membrane. The experimental assessment of flux criticality was carried out by flux-stepping tests showing the positive impact of liquid temperature on the value of the critical threshold. The reliability of short-term tests was then verified over a long period by determining the time of sustainability, t(sust), of six different sub-critical fluxes ranging between 17 and 30Lm(-2)h(-1). An exponential fitting was observed in terms of fouling rate both before and after t(sust), though fouling after t(sust) is likely to be ascribed not only to cake formation. Finally, a new mathematical formulation was proposed according to the local flux approach to model the sub-critical TMP transients. The model involves both bound and free forms of EPS and, once experimentally calibrated, it provided a fair prediction of the TMP jump.

  11. Studies on the effects of H{sub 2}O{sub 2} as an oxygen source in microbial decomposition of hydrocarbons in liquid culture and in near-to-natural bioreactors; Untersuchungen zu den Effekten von H{sub 2}O{sub 2} als Sauerstoffquelle beim mikrobiellen Abbau von Kohlenwasserstoffen in Fluessigkultur und in-s-situ-nahen Bodenreaktoren

    Energy Technology Data Exchange (ETDEWEB)

    Mueller, B.G.

    1992-08-12

    Hydrocarbon decomposition in soil with hydrogen peroxide as an oxygen source was examined. Among the model hydrocarbons was diesel fuel. Decomposition in liquid culture was studied in a bioreactor using mixed cultures. Hydrogen peroxide proved its value as an oxygen source. It was found that in soil systems hydrocarbons are largely mineralized and only small amounts of biomass are formed. (VT) [Deutsch] Untersucht wurde der Kohlenwasserstoffabbau im Boden unter Sauerstoffversorgung durch Wasserstoffperoxid. Als Modellkohlenwasserstoff wurde unter anderem Dieseloel eingesetzt. In einem Bioreaktor wurde der Abbau in Fluessigkultur durch Mischkulturen studiert. Wasserstoffperoxid hat sich als Sauerstoffversorger bewaehrt. In Bodensystemen stellte man fest, dass die Kohlenwasserstoffe weitgehend mineralisiert werden und nur wenig Biomasse gebildet wird. (VT)

  12. 甘草细胞在搅拌式生物反应器中的放大培养%Ampliifcation Culture of Glycyrrhiza uralensis Cell in Stirring Bioreactor

    Institute of Scientific and Technical Information of China (English)

    李雅丽; 孟婷婷; 王毛毛; 李晓雪; 李蓉蓉; 郭晓强

    2015-01-01

    Based on the establishment of a stable Glycyrrhiza uralensis (licorice) cells ampliifcation culture system in stirring bioreactor, the growth characteristics of cell suspension culture in stirring bioreactor were studied, including the cell growth, cell membrane permeability, the pH change of the culture system and licorice lfavonoids synthesis, and compared with the shake lfask culture. The results showed that cell biomass accumu-lation in the bioreactor was less than that in shake lfask under the same conditions, and the whole cultivation period was shortened. At the same time in the culture period, pH was slightly lower than that in the shake lfask, the H2O2 concentration in cells was 1.8 times of that in shake lfask, and the production of licorice lfavonoids was 1.5 times of that in the shake lfask. These indicated that the cultivation environment with mechanical agita-tion and lfuid shear in the bioreactor inhibited the cells growth at a certain level, but stimulated secondary me-tabolites licorice lfavonoids synthesis cell in higher levels.%在建立了稳定的甘草细胞搅拌式生物反应器放大培养体系的基础上,本文研究了甘草细胞在搅拌式反应器中悬浮培养的生长特性,包括细胞生长、细胞膜的透性、培养体系的pH变化及甘草黄酮合成情况等,并与摇瓶培养作比较。结果发现,同等条件下,反应器中培养细胞生物量的积累低于摇瓶培养,整个培养周期较摇瓶培养缩短。培养过程中同一时间段反应器中的pH值略低于摇瓶中的pH,细胞中H2O2的浓度是摇瓶中的1.8倍,甘草黄酮的产量是摇瓶培养的1.5倍,表明反应器中机械搅拌与流体剪切的培养环境对细胞生长起到一定程度的抑制作用,但刺激了细胞次生代谢产物甘草黄酮较高水平的合成。

  13. Bioreactor and methods for producing synchronous cells

    Science.gov (United States)

    Helmstetter, Charles E. (Inventor); Thornton, Maureen (Inventor); Gonda, Steve (Inventor)

    2005-01-01

    Apparatus and methods are directed to a perfusion culture system in which a rotating bioreactor is used to grow cells in a liquid culture medium, while these cells are attached to an adhesive-treated porous surface. As a result of this arrangement and its rotation, the attached cells divide, with one cell remaining attached to the substrate, while the other cell, a newborn cell is released. These newborn cells are of approximately the same age, that are collected upon leaving the bioreactor. The populations of newborn cells collected are of synchronous and are minimally, if at all, disturbed metabolically.

  14. Modelling of a Batch Whey Cultivation of Kluyveromyces marxianus var. lactis MC 5 with Investigation of Mass Transfer Processes in the Bioreactor

    OpenAIRE

    Mitko Petrov; Tatiana Ilkova; Juris Vanags

    2015-01-01

    This study presents a mathematical model of a batch fermentation of lactose oxidation from a natural substratum in a cultivation by the strain Kluyweromyces marxianus var. lactis MC 5. In the model of the process, the mass transfer in the bioreactor for oxygen concentration in the gas phase (GP) and in the liquid phase (LP) is based on the dispersion model of the GP. In addition, perfect mixing in LP is included. Nine models were investigated for specific growth rate and specific oxygen consu...

  15. Rotating cell culture systems for human cell culture: human trophoblast cells as a model.

    Science.gov (United States)

    Zwezdaryk, Kevin J; Warner, Jessica A; Machado, Heather L; Morris, Cindy A; Höner zu Bentrup, Kerstin

    2012-01-18

    The field of human trophoblast research aids in understanding the complex environment established during placentation. Due to the nature of these studies, human in vivo experimentation is impossible. A combination of primary cultures, explant cultures and trophoblast cell lines support our understanding of invasion of the uterine wall and remodeling of uterine spiral arteries by extravillous trophoblast cells (EVTs), which is required for successful establishment of pregnancy. Despite the wealth of knowledge gleaned from such models, it is accepted that in vitro cell culture models using EVT-like cell lines display altered cellular properties when compared to their in vivo counterparts. Cells cultured in the rotating cell culture system (RCCS) display morphological, phenotypic, and functional properties of EVT-like cell lines that more closely mimic differentiating in utero EVTs, with increased expression of genes mediating invasion (e.g. matrix metalloproteinases (MMPs)) and trophoblast differentiation. The Saint Georges Hospital Placental cell Line-4 (SGHPL-4) (kindly donated by Dr. Guy Whitley and Dr. Judith Cartwright) is an EVT-like cell line that was used for testing in the RCCS. The design of the RCCS culture vessel is based on the principle that organs and tissues function in a three-dimensional (3-D) environment. Due to the dynamic culture conditions in the vessel, including conditions of physiologically relevant shear, cells grown in three dimensions form aggregates based on natural cellular affinities and differentiate into organotypic tissue-like assemblies. The maintenance of a fluid orbit provides a low-shear, low-turbulence environment similar to conditions found in vivo. Sedimentation of the cultured cells is countered by adjusting the rotation speed of the RCCS to ensure a constant free-fall of cells. Gas exchange occurs through a permeable hydrophobic membrane located on the back of the bioreactor. Like their parental tissue in vivo, RCCS

  16. Immersed Boundary Models for Quantifying Flow-Induced Mechanical Stimuli on Stem Cells Seeded on 3D Scaffolds in Perfusion Bioreactors

    Science.gov (United States)

    Smeets, Bart; Odenthal, Tim; Luyten, Frank P.; Ramon, Herman; Papantoniou, Ioannis; Geris, Liesbet

    2016-01-01

    Perfusion bioreactors regulate flow conditions in order to provide cells with oxygen, nutrients and flow-associated mechanical stimuli. Locally, these flow conditions can vary depending on the scaffold geometry, cellular confluency and amount of extra cellular matrix deposition. In this study, a novel application of the immersed boundary method was introduced in order to represent a detailed deformable cell attached to a 3D scaffold inside a perfusion bioreactor and exposed to microscopic flow. The immersed boundary model permits the prediction of mechanical effects of the local flow conditions on the cell. Incorporating stiffness values measured with atomic force microscopy and micro-flow boundary conditions obtained from computational fluid dynamics simulations on the entire scaffold, we compared cell deformation, cortical tension, normal and shear pressure between different cell shapes and locations. We observed a large effect of the precise cell location on the local shear stress and we predicted flow-induced cortical tensions in the order of 5 pN/μm, at the lower end of the range reported in literature. The proposed method provides an interesting tool to study perfusion bioreactors processes down to the level of the individual cell’s micro-environment, which can further aid in the achievement of robust bioprocess control for regenerative medicine applications. PMID:27658116

  17. Loading and Boundary Condition Influences in a Poroelastic Finite Element Model of Cartilage Stresses in a Triaxial Compression Bioreactor

    Science.gov (United States)

    Kallemeyn, Nicole A; Grosland, Nicole M; Pedersen, Doug R; Martin, James A; Brown, Thomas D

    2006-01-01

    Background: We developed a poroelastic finite element (FE) model of cartilage in dynamic triaxial compression to parametrically analyze the effects of loading and boundary conditions on a baseline model. Conventional mechanical tests on articular cartilage such as confined and unconfined compression, indentation, etc., do not fully allow for modulation of compression and shear at physiological levels whereas triaxial compression does. A Triaxial Compression Bioreactor, or TRIAX, has been developed to study chondrocyte responses to multi-axial stress conditions under cyclic loading. In the triaxial setting, however, a cartilage explant's physical testing environment departs from the ideal homogeneous stress state that would occur from strict linear superposition of the applied axial and transverse pressure. Method of Approach: An axisymmetric poroelastic FE model of a cartilage explant (4 mm diameter, 1.5 mm thick) in cyclic triaxial compression was created. Axial and transverse loads (2 MPa at 1 Hz.) were applied via a platen and containment sheath. Parameters of interest included the rise time and magnitude of the applied load, in addition to the containment sheath modulus and the friction coefficient at the cartilage/platen interfaces. Metrics of interest in addition to whole explant axial strain included axial (surface normal) stress, shear stress, pore pressure, and the fluid load carriage fraction within the explant. Results: Strain results were compared to experimental data from explants tested in the TRIAX under conditions similar to the baseline model. Explant biomechanics varied considerably over numbers of load cycles and parameter values. Cyclic loading caused an increase in accumulated strain for the various loading and boundary conditions. Conclusions: Unlike what would be expected from linear superposition of the homogeneous stresses from the applied axial and transverse pressure, we have shown that the stress state within the TRIAX is considerably

  18. Organizational culture diagnosis - a new model

    Directory of Open Access Journals (Sweden)

    Ph. D. Ionuţ Constantin

    2010-05-01

    Full Text Available Organizational culture is a key source of competitive advantage. There is a demonstrated relation between organizational culture and organizational performance. This paper reviews previous research in the field and introduce a new model for understanding, diagnosing and changing organizational culture. The main advantage of the new model is based on regarding culture as the management and work practices that are either hindering or helping an organization's bottom line performance.

  19. Safety Cultural Competency Modeling in Nuclear Organizations

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Sa Kil; Oh, Yeon Ju; Luo, Meiling; Lee, Yong Hee [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

    2014-05-15

    The nuclear safety cultural competency model should be supplemented through a bottom-up approach such as behavioral event interview. The developed model, however, is meaningful for determining what should be dealt for enhancing safety cultural competency of nuclear organizations. The more details of the developing process, results, and applications will be introduced later. Organizational culture include safety culture in terms of its organizational characteristics.

  20. Feasibility and simulation model of a pilot scale membrane bioreactor for wastewater treatment and reuse from Chinese traditional medicine

    Institute of Scientific and Technical Information of China (English)

    REN Nan-qi; YAN Xian-feng; CHEN Zhao-bo; HU Dong-xue; GONG Man-li; GUO Wan-qian

    2007-01-01

    The lack and pollution of water resource make wastewater reuse necessary. The pilot scale long-term tests for submerged membrane bioreactor were conducted to treat the effluents of anaerobic or aerobic treatment process for the high-strength Chinese traditional medicine wastewater. This article was focused on the feasibility of the wastewater treatment and reuse at shorter hydraulic retention time (HRT) of 5.0, 3.2 and 2.13 h. MLSS growth, membrane flux, vacuum values and chemical cleaning periods were also investigated.The experimental results of treating two-phase anaerobic treatment effluent demonstrated that the CODfilt was less than 100 mg/L when the influent COD was between 500-10000 mg/L at HRT of 5.0 h, which could satisfy the normal discharged standard in China. The experimental results to treat cross flow aerobic reactor effluent demonstrated that the average value of CODfilt was 17.28 mg/L when the average value of infiuent COD was 192.84 mg/L at HRT of 2.13 h during 106 d, which could completely meet the normal standard for water reuse. The maximum MLSS and MLVSS reached 24000 and 14500 mg/L at HRT of 3.2 h respectively. Membrane flux had maximal resume degrees of 94.7% at vacuum value of 0.02 MPa after cleaning. Chemical cleaning periods of membrane module were 150 d. A simulation model of operational parameters was also established based on the theory of back propagation neural network and linear regression of traditional mathematical model. The simulation model showed that the optimum operational parameters were of MLSS was between 7543-13694 mg/L.

  1. Effect of bottom clearance on performance of airlift bioreactor in high-density culture of Panax notoginseng cells.

    Science.gov (United States)

    Hu, W; Zhong, J

    2001-01-01

    A fed-batch cultivation of Panax notoginseng cells in a concentric-tube airlift reactor was performed to study the effects of bottom clearance on cell growth and the production of ginseng saponin and polysaccharide. At a bottom clearance of 4.0 cm, the highest cell density of 29.1+/-1.6 g/l by dry weight was obtained, and the accumulation of saponin and polysaccharide also reached a maximum, i.e., 2.39+/-0.43 and 2.73+/-0.40 g/l, respectively. Cell growth and metabolite production were limited at a small (2.5 cm) or large (5.0 cm) bottom clearance. By analyzing the time constants of mixing, mass transfer and oxygen consumption, bulk gas-liquid oxygen transfer was found to be responsible for the growth limitation at a small bottom clearance (2.5 cm). The decrease in cell density at a large bottom clearance (5.0 cm) was related to cell sedimentation at the reactor bottom. This work is beneficial for the scale-up and efficient operation of the airlift reactor in cell cultures.

  2. Mechanical Stimulus Inhibits the Growth of a Bone Tissue Model Cultured In Vitro

    Institute of Scientific and Technical Information of China (English)

    Zong-ming Wan; Lu Liu; Jian-yu Li; Rui-xin Li; Yong Guo; Hao Li; Jian-ming Zhang; Xi-zheng Zhang

    2013-01-01

    Objectives To construct the cancellous bone explant model and a method of culturing these bone tissues in vitro, and to investigate the effect of mechanical load on growth of cancellous bone tissue in vitro. Methods Cancellous bone were extracted from rabbit femoral head and cut into 1-mm-thick and 8-mm-diameter slices under sterile conditions. HE staining and scanning electron microscopy were employed to identify the histomorphology of the model after being cultured with a new dynamic load and circulating perfusion bioreactor system for 0, 3, 5, and 7 days, respectively. We built a three-dimensional model using microCT and analyzed the loading effects using finite element analysis. The model was subjected to mechanical load of 1000, 2000, 3000, and 4000μεrespectively for 30 minutes per day. After 5 days of continuous stimuli, the activities of alkaline phosphatase (AKP) and tartrate-resistant acid phosphatase (TRAP) were detected. Apoptosis was analyzed by DNA ladder detection and caspase-3/8/9 activity detection. Results After being cultured for 3, 5, and 7 days, the bone explant model grew well. HE staining showed the apparent nucleus in cells at the each indicated time, and electron microscope revealed the living cells in the bone tissue. The activities of AKP and TRAP in the bone explant model under mechanical load of 3000 and 4000μεwere significantly lower than those in the unstressed bone tissues (all P Conclusions The cancellous bone explant model extracted from the rabbit femoral head could be alive at least for 7 days in the dynamic load and circulating perfusion bioreactor system, however, pathological mechanical load could affect the bone tissue growth by apoptosis in vitro. The differentiation of osteoblasts and osteoclasts might be inhibited after the model is stimulated by mechanical load of 3000 and 4000με.

  3. A Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells.

    Science.gov (United States)

    Drummond, Coyne G; Nickerson, Cheryl A; Coyne, Carolyn B

    2016-01-01

    Despite serving as the primary entry portal for coxsackievirus B (CVB), little is known about CVB infection of the intestinal epithelium, owing at least in part to the lack of suitable in vivo models and the inability of cultured cells to recapitulate the complexity and structure associated with the gastrointestinal (GI) tract. Here, we report on the development of a three-dimensional (3-D) organotypic cell culture model of Caco-2 cells to model CVB infection of the gastrointestinal epithelium. We show that Caco-2 cells grown in 3-D using the rotating wall vessel (RWV) bioreactor recapitulate many of the properties of the intestinal epithelium, including the formation of well-developed tight junctions, apical-basolateral polarity, brush borders, and multicellular complexity. In addition, transcriptome analyses using transcriptome sequencing (RNA-Seq) revealed the induction of a number of genes associated with intestinal epithelial differentiation and/or intestinal processes in vivo when Caco-2 cells were cultured in 3-D. Applying this model to CVB infection, we found that although the levels of intracellular virus production were similar in two-dimensional (2-D) and 3-D Caco-2 cell cultures, the release of infectious CVB was enhanced in 3-D cultures at early stages of infection. Unlike CVB, the replication of poliovirus (PV) was significantly reduced in 3-D Caco-2 cell cultures. Collectively, our studies show that Caco-2 cells grown in 3-D using the RWV bioreactor provide a cell culture model that structurally and transcriptionally represents key aspects of cells in the human GI tract and can thus be used to expand our understanding of enterovirus-host interactions in intestinal epithelial cells. IMPORTANCE Coxsackievirus B (CVB), a member of the enterovirus family of RNA viruses, is associated with meningitis, pericarditis, diabetes, dilated cardiomyopathy, and myocarditis, among other pathologies. CVB is transmitted via the fecal-oral route and encounters the

  4. In vivo bioreactors for mandibular reconstruction.

    Science.gov (United States)

    Tatara, A M; Wong, M E; Mikos, A G

    2014-12-01

    Large mandibular defects are difficult to reconstruct with good functional and aesthetic outcomes because of the complex geometry of craniofacial bone. While the current gold standard is free tissue flap transfer, this treatment is limited in fidelity by the shape of the harvested tissue and can result in significant donor site morbidity. To address these problems, in vivo bioreactors have been explored as an approach to generate autologous prefabricated tissue flaps. These bioreactors are implanted in an ectopic site in the body, where ossified tissue grows into the bioreactor in predefined geometries and local vessels are recruited to vascularize the developing construct. The prefabricated flap can then be harvested with vessels and transferred to a mandibular defect for optimal reconstruction. The objective of this review article is to introduce the concept of the in vivo bioreactor, describe important preclinical models in the field, summarize the human cases that have been reported through this strategy, and offer future directions for this exciting approach.

  5. 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.

  6. Bridging the gap between traditional cell cultures and bioreactors applied in regenerative medicine: practical experiences with the MINUSHEET perfusion culture system.

    Science.gov (United States)

    Minuth, Will W; Denk, Lucia

    2016-03-01

    To meet specific requirements of developing tissues urgently needed in tissue engineering, biomaterial research and drug toxicity testing, a versatile perfusion culture system was developed. First an individual biomaterial is selected and then mounted in a MINUSHEET(®) tissue carrier. After sterilization the assembly is transferred by fine forceps to a 24 well culture plate for seeding cells or mounting tissue on it. To support spatial (3D) development a carrier can be placed in various types of perfusion culture containers. In the basic version a constant flow of culture medium provides contained tissue with always fresh nutrition and respiratory gas. For example, epithelia can be transferred to a gradient container, where they are exposed to different fluids at the luminal and basal side. To observe development of tissue under the microscope, in a different type of container a transparent lid and base are integrated. Finally, stem/progenitor cells are incubated in a container filled by an artificial interstitium to support spatial development. In the past years the described system was applied in numerous own and external investigations. To present an actual overview of resulting experimental data, the present paper was written.

  7. A Dynamic Model of Cultural Reproduction

    DEFF Research Database (Denmark)

    Jæger, Mads Meier; Breen, Richard

    2016-01-01

    of study. Their model describes how parents transmit cultural capital to their children and how children convert cultural capital into educational success. It also provides a behavioral framework for interpreting parental investments in cultural capital. The authors review results from existing empirical......The authors draw on Pierre Bourdieu’s theory of cultural reproduction to develop a formal model of the pathways through which cultural capital acts to enhance children’s educational and socioeconomic success. The authors’ approach brings conceptual and empirical clarity to an important area...

  8. A novel dual-flow bioreactor simulates increased fluorescein permeability in epithelial tissue barriers.

    Science.gov (United States)

    Giusti, Serena; Sbrana, Tommaso; La Marca, Margherita; Di Patria, Valentina; Martinucci, Valentina; Tirella, Annalisa; Domenici, Claudio; Ahluwalia, Arti

    2014-09-01

    Permeability studies across epithelial barriers are of primary importance in drug delivery as well as in toxicology. However, traditional in vitro models do not adequately mimic the dynamic environment of physiological barriers. Here, we describe a novel two-chamber modular bioreactor for dynamic in vitro studies of epithelial cells. The fluid dynamic environment of the bioreactor was characterized using computational fluid dynamic models and measurements of pressure gradients for different combinations of flow rates in the apical and basal chambers. Cell culture experiments were then performed with fully differentiated Caco-2 cells as a model of the intestinal epithelium, comparing the effect of media flow applied in the bioreactor with traditional static transwells. The flow increases barrier integrity and tight junction expression of Caco-2 cells with respect to the static controls. Fluorescein permeability increased threefold in the dynamic system, indicating that the stimulus induced by flow increases transport across the barrier, closely mimicking the in vivo situation. The results are of interest for studying the influence of mechanical stimuli on cells, and underline the importance of developing more physiologically relevant in vitro tissue models. The bioreactor can be used to study drug delivery, chemical, or nanomaterial toxicity and to engineer barrier tissues.

  9. Optimizing of Bioreactor Heat Supply and Material Feeding by Numerical Calculation

    Science.gov (United States)

    Zhou, Zhiwei; Song, Boyan; Zhu, Likuan; Li, Zuntao; Wang, Yang

    Cell culture at large scale normally uses stirred structure. And the situation of temperature field distribution is very important to the cell culture at large scale. Some cells are very sensitive to the circumstances. The local temperature is too high or too low all influences the cell survival and low the cell quantity at unit volume. This paper simulates the temperature field under three different heating conditions. Then analysis and contrast the simulation results. The mixed situation in bioreactor is extremely significant for nutrition transmit. Usually, use ways to measure the average mixture time in bioreactor, and improve the mixture circumstance in the bioreactor through stirred impeller and bioreactor structure change. This paper adopts numerical calculation method to investigate the flow field in bioreactor. It gets the mixture time of bioreactor through virtual tracer in simulate flow field and detects the tracer density time variation curve in the bioreactor.

  10. Bioreactor Mass Transport Studies

    Science.gov (United States)

    Kleis, Stanley J.; Begley, Cynthia M.

    1997-01-01

    The objectives of the proposed research efforts were to develop both a simulation tool and a series of experiments to provide a quantitative assessment of mass transport in the NASA rotating wall perfused vessel (RWPV) bioreactor to be flown on EDU#2. This effort consisted of a literature review of bioreactor mass transport studies, the extension of an existing scalar transport computer simulation to include production and utilization of the scalar, and the evaluation of experimental techniques for determining mass transport in these vessels. Since mass transport at the cell surface is determined primarily by the relative motion of the cell assemblage and the surrounding fluid, a detailed assessment of the relative motion was conducted. Results of the simulations of the motion of spheres in the RWPV under microgravity conditions are compared with flight data from EDU#1 flown on STS-70. The mass transport across the cell membrane depends upon the environment, the cell type, and the biological state of the cell. Results from a literature review of cell requirements of several scalars are presented. As a first approximation, a model with a uniform spatial distribution of utilization or production was developed and results from these simulations are presented. There were two candidate processes considered for the experimental mass transport evaluations. The first was to measure the dissolution rate of solid or gel beads. The second was to measure the induced fluorescence of beads as a stimulant (for example hydrogen peroxide) is infused into the vessel. Either technique would use video taped images of the process for recording the quantitative results. Results of preliminary tests of these techniques are discussed.

  11. Modeling of a trickling bioreactor for the simultaneous removal of hydrogen sulfide and carbon disulfide

    Energy Technology Data Exchange (ETDEWEB)

    Lobo, R.; Viveros-Garcia, T.; Revah, S. [Universidad Autonoma Metropolitana-Iztapalapa, Mexico D.F. (Mexico)

    1996-12-31

    A model is presented for a trickle biofilm reactor based on liquid falling film hydrodynamics and linear residence times. It is shown that mass transfer resistance is high. Performance of pseudo homogeneous and heterogeneous models is discussed. It is shown that the use of superficial residence times reduces the heterogeneous model to a pseudo homogeneous one and masks parametric response. 8 refs., 2 figs., 1 tab.

  12. High cell density cultivation of recombinant yeasts and bacteria under non-pressurized and pressurized conditions in stirred tank bioreactors.

    Science.gov (United States)

    Knoll, Arnd; Bartsch, Stefan; Husemann, Bernward; Engel, Philip; Schroer, Kirsten; Ribeiro, Betina; Stöckmann, Christoph; Seletzky, Juri; Büchs, Jochen

    2007-10-31

    This study demonstrates the applicability of pressurized stirred tank bioreactors for oxygen transfer enhancement in aerobic cultivation processes. The specific power input and the reactor pressure was employed as process variable. As model organism Escherichia coli, Arxula adeninivorans, Saccharomyces cerevisiae and Corynebacterium glutamicum were cultivated to high cell densities. By applying specific power inputs of approx. 48kWm(-3) the oxygen transfer rate of a E. coli culture in the non-pressurized stirred tank bioreactor was lifted up to values of 0.51moll(-1)h(-1). When a reactor pressure up to 10bar was applied, the oxygen transfer rate of a pressurized stirred tank bioreactor was lifted up to values of 0.89moll(-1)h(-1). The non-pressurized stirred tank bioreactor was able to support non-oxygen limited growth of cell densities of more than 40gl(-1) cell dry weight (CDW) of E. coli, whereas the pressurized stirred tank bioreactor was able to support non-oxygen limited growth of cell densities up to 225gl(-1) CDW of A. adeninivorans, 89gl(-1) CDW of S. cerevisiae, 226gl(-1) CDW of C. glutamicum and 110gl(-1) CDW of E. coli. Compared to literature data, some of these cell densities are the highest values ever achieved in high cell density cultivation of microorganisms in stirred tank bioreactors. By comparing the specific power inputs as well as the k(L)a values of both systems, it is demonstrated that only the pressure is a scaleable tool for oxygen transfer enhancement in industrial stirred tank bioreactors. Furthermore, it was shown that increased carbon dioxide partial pressures did not remarkably inhibit the growth of the investigated model organisms.

  13. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  15. Hydrodynamics research of wastewater treatment bioreactors

    Institute of Scientific and Technical Information of China (English)

    REN Nan-qi; ZHANG Bing; ZHOU Xue-fei

    2009-01-01

    To optimize the design and improve the performance of wastewater treatment bioreactors, the review concerning the hydrodynamics explored by theoretical equations, process experiments, modeling of the hydrody-namics and flow field measurement is presented. Results of different kinds of experiments show that the hydro-dynamic characteristics can affect sludge characteristics, mass transfer and reactor performance significantly. A-long with the development of theoretical equations, turbulence models including large eddy simulation models and Reynolds-averaged Navier-Stokes (RANS) models are widely used at present. Standard and modified k-ε models are the most widely used eddy viscosity turbulence models for simulating flows in bioreactors. Numericalsimulation of hydrodynamics is proved to be efficient for optimizing design and operation. The development of measurement techniques with high accuracy and low intrusion enables the flow filed in the bioreactors to be transparent. Integration of both numerical simulation and experimental measurement can describe the hydrody-namics very well.

  16. Modeling of the pyruvate production with Escherichia coli in a fed-batch bioreactor.

    Science.gov (United States)

    Zelić, B; Vasić-Racki, D; Wandrey, C; Takors, R

    2004-07-01

    A family of 10 competing, unstructured models has been developed to model cell growth, substrate consumption, and product formation of the pyruvate producing strain Escherichia coli YYC202 ldhA::Kan strain used in fed-batch processes. The strain is completely blocked in its ability to convert pyruvate into acetyl-CoA or acetate (using glucose as the carbon source) resulting in an acetate auxotrophy during growth in glucose minimal medium. Parameter estimation was carried out using data from fed-batch fermentation performed at constant glucose feed rates of q(VG)=10 mL h(-1). Acetate was fed according to the previously developed feeding strategy. While the model identification was realized by least-square fit, the model discrimination was based on the model selection criterion (MSC). The validation of model parameters was performed applying data from two different fed-batch experiments with glucose feed rate q(VG)=20 and 30 mL h(-1), respectively. Consequently, the most suitable model was identified that reflected the pyruvate and biomass curves adequately by considering a pyruvate inhibited growth (Jerusalimsky approach) and pyruvate inhibited product formation (described by modified Luedeking-Piret/Levenspiel term).

  17. Validation of computational non-Newtonian fluid model for membrane bioreactor

    DEFF Research Database (Denmark)

    Sørensen, Lasse; Bentzen, Thomas Ruby; Skov, Kristian

    2015-01-01

    for optimizing MBR-systems is computational fluid dynamics (CFD) modelling, giving the ability to describe the flow in the systems. A parameter which is often neglected in such models is the non-Newtonian properties of active sludge, which is of great importance for MBR systems since they operate at sludge...... concentrations up to a factor 10 compared to conventional activated sludge (CAS) systems, resulting in strongly shear thinning liquids. A CFD-model is validated against measurements conducted in a system with rotating cross flow membranes submerged in non-Newtonian liquids, where tangential velocities...

  18. 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

    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. PMID:24967717

  19. Bioreactor Scalability: Laboratory-Scale Bioreactor Design Influences Performance, Ecology, and Community Physiology in Expanded Granular Sludge Bed Bioreactors.

    Science.gov (United States)

    Connelly, Stephanie; Shin, Seung G; Dillon, Robert J; Ijaz, Umer Z; Quince, Christopher; Sloan, William T; Collins, Gavin

    2017-01-01

    Studies investigating the feasibility of new, or improved, biotechnologies, such as wastewater treatment digesters, inevitably start with laboratory-scale trials. However, it is rarely determined whether laboratory-scale results reflect full-scale performance or microbial ecology. The Expanded Granular Sludge Bed (EGSB) bioreactor, which is a high-rate anaerobic digester configuration, was used as a model to address that knowledge gap in this study. Two laboratory-scale idealizations of the EGSB-a one-dimensional and a three- dimensional scale-down of a full-scale design-were built and operated in triplicate under near-identical conditions to a full-scale EGSB. The laboratory-scale bioreactors were seeded using biomass obtained from the full-scale bioreactor, and, spent water from the distillation of whisky from maize was applied as substrate at both scales. Over 70 days, bioreactor performance, microbial ecology, and microbial community physiology were monitored at various depths in the sludge-beds using 16S rRNA gene sequencing (V4 region), specific methanogenic activity (SMA) assays, and a range of physical and chemical monitoring methods. SMA assays indicated dominance of the hydrogenotrophic pathway at full-scale whilst a more balanced activity profile developed during the laboratory-scale trials. At each scale, Methanobacterium was the dominant methanogenic genus present. Bioreactor performance overall was better at laboratory-scale than full-scale. We observed that bioreactor design at laboratory-scale significantly influenced spatial distribution of microbial community physiology and taxonomy in the bioreactor sludge-bed, with 1-D bioreactor types promoting stratification of each. In the 1-D laboratory bioreactors, increased abundance of Firmicutes was associated with both granule position in the sludge bed and increased activity against acetate and ethanol as substrates. We further observed that stratification in the sludge-bed in 1-D laboratory

  20. MATHEMATICAL MODELLING FOR THE CONVERSION OF ANIMAL WASTE TO METHANE IN BATCH BIOREACTOR

    Directory of Open Access Journals (Sweden)

    O.A. Aworanti

    2011-01-01

    Full Text Available An investigation was conducted to predict the behaviour of microbial processes leading to the production of biogas from animal waste. Mathematical model were developed for the prediction of the behaviour of microbial processes. The development of the models was based upon a material balance analysis of the digester operation, substrate utilization, cell growth and product formation. The model was solved using Runge kutta numerical technique embedded in polymath software. The digesters’ operations simulated with a starting valve of 300g/dm3 as the concentration of the substrate and 1.5g/dm3 as the concentration of the cell, within a period of 13days. The results of the simulation show that the substrate concentration shows exponential decline from (300g/dm3 to 6.88g/dm3, the cells growth shows exponential trend from (1.5g/dm3to 39g/dm3 The rate of growth of cell was increased from (0.5g/dm3-2.53g/dm3, death increased from (0.015g/dm3 to 0.161g/dm3 over the 13days and the biogas production which is the product also follow the exponential trend from (zeroconcentration to 219g/dm3. In all the model does the prediction well on all the parameters simulated, so it was can be used to predict the product formation rate as well as the design of reactor or digester.

  1. Tapered bed bioreactor

    Science.gov (United States)

    Scott, Charles D.; Hancher, Charles W.

    1977-01-01

    A vertically oriented conically shaped column is used as a fluidized bed bioreactor wherein biologically catalyzed reactions are conducted in a continuous manner. The column utilizes a packing material a support having attached thereto a biologically active catalytic material.

  2. Inferring cultural models from corpus data

    DEFF Research Database (Denmark)

    Jensen, Kim Ebensgaard

    2015-01-01

    developed methods of inferring cultural models from observed behavior – in particular observed verbal behavior (including both spoken and written language). While there are plenty of studies of the reflection of cultural models in artificially generated verbal behavior, not much research has been made...... of constructional discursive behavior, the present paper offers a covarying collexeme analysis of the [too ADJ to V]-construction in the Corpus of Contemporary American English. The purpose is to discover the extent to which its force-dynamic constructional semantics interacts with cultural models. We focus...

  3. Hydrodynamic characteristics of UASB bioreactors.

    Science.gov (United States)

    John, Siby; Tare, Vinod

    2011-10-01

    The hydrodynamic characteristics of UASB bioreactors operated under different organic loading and hydraulic loading rates were studied, using three laboratory scale models treating concocted sucrose wastewater. Residence time distribution (RTD) analysis using dispersion model and tanks-in-series model was directed towards the characterization of the fluid flow pattern in the reactors and correlation of the hydraulic regime with the biomass content and biogas production. Empty bed reactors followed a plug flow pattern and the flow pattern changed to a large dispersion mixing with biomass and gas production. Effect of increase in gas production on the overall hydraulics was insignificant.

  4. A Dynamic Model of Cultural Reproduction

    DEFF Research Database (Denmark)

    Jæger, Mads Meier; Breen, Richard

    2016-01-01

    The authors draw on Pierre Bourdieu’s theory of cultural reproduction to develop a formal model of the pathways through which cultural capital acts to enhance children’s educational and socioeconomic success. The authors’ approach brings conceptual and empirical clarity to an important area of st...

  5. An innovative lattice Boltzmann model for simulating Michaelis-Menten-based diffusion-advection kinetics and its application within a cartilage cell bioreactor.

    Science.gov (United States)

    Moaty Sayed, A A; Hussein, M A; Becker, T

    2010-04-01

    Lattice Boltzmann models (LBM) are rapidly showing their ability to simulate a lot of fluid dynamics problems that previously required very complex approaches. This study presents a LBM for simulating diffusion-advection transport of substrate in a 2-D laminar flow. The model considers the substrate influx into a set of active cells placed inside the flow field. A new innovative method was used to simulate the cells activity using the LBM by means of Michaelis-Menten kinetics. The model is validated with some numerical benchmark problems and proved highly accurate results. After validation the model was used to simulate the transport of oxygen substrates that diffuse in water to feed a set of active cartilage cells inside a new designed bioreactor.

  6. Multi-Level Cultural Models

    Science.gov (United States)

    2014-11-05

    people, groups, ideas and beliefs in a complex web of relations that changes through time and space. If we are to explain and predict socio-cultural... scraped from Lexis-Nexis covering 16 countries for 20 months from August 2010 through March 2012. This data was segmented by country by month...country a generalized other was added to represent the general public and this was linked to the average of the real data. Using web -based reviews

  7. Bioreactor Studies and Computational Fluid Dynamics

    Science.gov (United States)

    Singh, H.; Hutmacher, D. W.

    The hydrodynamic environment “created” by bioreactors for the culture of a tissue engineered construct (TEC) is known to influence cell migration, proliferation and extra cellular matrix production. However, tissue engineers have looked at bioreactors as black boxes within which TECs are cultured mainly by trial and error, as the complex relationship between the hydrodynamic environment and tissue properties remains elusive, yet is critical to the production of clinically useful tissues. It is well known in the chemical and biotechnology field that a more detailed description of fluid mechanics and nutrient transport within process equipment can be achieved via the use of computational fluid dynamics (CFD) technology. Hence, the coupling of experimental methods and computational simulations forms a synergistic relationship that can potentially yield greater and yet, more cohesive data sets for bioreactor studies. This review aims at discussing the rationale of using CFD in bioreactor studies related to tissue engineering, as fluid flow processes and phenomena have direct implications on cellular response such as migration and/or proliferation. We conclude that CFD should be seen by tissue engineers as an invaluable tool allowing us to analyze and visualize the impact of fluidic forces and stresses on cells and TECs.

  8. Model-based analysis and design of a microchannel reactor for tissue engineering.

    Science.gov (United States)

    Mehta, Khamir; Linderman, Jennifer J

    2006-06-20

    Recently developed perfusion micro-bioreactors offer the promise of more physiologic in vitro systems for tissue engineering. Successful application of such bioreactors will require a method to characterize the bioreactor environment required to elicit desired cell function. We present a mathematical model to describe nutrient/growth factor transport and cell growth inside a microchannel bioreactor. Using the model, we first show that the nature of spatial gradients in nutrient concentration can be controlled by both design and operating conditions and are a strong function of cell uptake rates. Next, we extend our model to investigate the spatial distributions of cell-secreted soluble autocrine/paracrine growth factors in the bioreactor. We show that the convective transport associated with the continuous cell culture and possible media recirculation can significantly alter the concentration distribution of the soluble signaling molecules as compared to static culture experiments and hence needs special attention when adapting static culture protocols for the bioreactor. Further, using an unsteady state model, we find that spatial gradients in nutrient/growth factor concentrations can bring about spatial variations in the cell density distribution inside the bioreactor, which can result in lowered working volume of the bioreactor. Finally, we show that the nutrient and spatial limitations can dramatically affect the composition of a co-cultured cell population. Our results are significant for the development, design, and optimization of novel micro-channel systems for tissue engineering.

  9. 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.

  10. Design and validation of a corneal bioreactor.

    Science.gov (United States)

    Leonard, Elissa K; Pai, Vincent H; Amberg, Philip; Gardner, Jens; Orwin, Elizabeth J

    2012-12-01

    Mechanical strain is an important signal that influences the behavior and properties of cells in a wide variety of tissues. Physiologically similar mechanical strain can revert cultured cells to a more normal phenotype. Here, we have demonstrated that 3% equibiaxial (EB) and uniaxial strains confer favorable protein expression in cultured rabbit corneal fibroblasts (RCFs), with approximately 35% and 65% reduction in expression of α-smooth muscle actin (α-SMA), respectively. We have designed a novel bioreactor that is capable of imparting up to 7% EB strain and up to 6% EB strain using a cornea-shaped post. Additional features of the bioreactor include the application of shear stress to cells in culture and the ability to image cells using optical coherence microscopy (OCM) without being removed from the system. Copyright © 2012 Wiley Periodicals, Inc.

  11. Galvanotactic migration of EA.Hy926 endothelial cells in a novel designed electric field bioreactor.

    Science.gov (United States)

    Long, Haiyan; Yang, Gang; Wang, Zhengrong

    2011-12-01

    Endogenous direct current electric fields (dcEFs) play a significant role in major biological processes such as embryogenesis, wound healing, and tissue regeneration. In this study, the galvanotaxis of human umbilical vein endothelial cell line EA.Hy926 was investigated by using a novel designed bioreactor. The physical features of the bioreactor were discussed and analyzed by both numerical simulation method and equivalent circuit model method. EA.Hy926 cells were cultured in the bioreactor for 10-24 h under 50-250 mV/mm dcEFs. Cell migration direction, distance, and velocity were recorded under an online time-lapse microscope. The effects of serum and growth factor on cell galvanotatic migration were investigated. To further explore the role of dcEFs in regulating endothelial cells, we analyzed the endothelial cell proliferation and secretion of nitric oxide (NO), endothelin-1 (ET-1) in response to dcEFs of physiological strength. Our results showed that EA.Hy926 cells had an obvious directional migration to the cathode, and the EF-directed migration was voltage dependent. The results also showed dcEFs did not affect cell proliferation, but affected the productions of NO and ET-1. Our study also showed the novel bioreactor, with a compact and planar style, makes it more convenient and more reasonable for EF stimulation experiments than earlier chamber designs.

  12. Dual-Purpose Bioreactors to Monitor Noninvasive Physical and Biochemical Markers of Kidney and Liver Scaffold Recellularization.

    Science.gov (United States)

    Uzarski, Joseph S; Bijonowski, Brent M; Wang, Bo; Ward, Heather H; Wandinger-Ness, Angela; Miller, William M; Wertheim, Jason A

    2015-10-01

    Analysis of perfusion-based bioreactors for organ engineering and a detailed evaluation of physical and biochemical parameters that measure dynamic changes within maturing cell-laden scaffolds are critical components of ex vivo tissue development that remain understudied topics in the tissue and organ engineering literature. Intricately designed bioreactors that house developing tissue are critical to properly recapitulate the in vivo environment, deliver nutrients within perfused media, and monitor physiological parameters of tissue development. Herein, we provide an in-depth description and analysis of two dual-purpose perfusion bioreactors that improve upon current bioreactor designs and enable comparative analyses of ex vivo scaffold recellularization strategies and cell growth performance during long-term maintenance culture of engineered kidney or liver tissues. Both bioreactors are effective at maximizing cell seeding of small-animal organ scaffolds and maintaining cell survival in extended culture. We further demonstrate noninvasive monitoring capabilities for tracking dynamic changes within scaffolds as the native cellular component is removed during decellularization and model human cells are introduced into the scaffold during recellularization and proliferate in maintenance culture. We found that hydrodynamic pressure drop (ΔP) across the retained scaffold vasculature is a noninvasive measurement of scaffold integrity. We further show that ΔP, and thus resistance to fluid flow through the scaffold, decreases with cell loss during decellularization and correspondingly increases to near normal values for whole organs following recellularization of the kidney or liver scaffolds. Perfused media may be further sampled in real time to measure soluble biomarkers (e.g., resazurin, albumin, or kidney injury molecule-1) that indicate degree of cellular metabolic activity, synthetic function, or engraftment into the scaffold. Cell growth within bioreactors is

  13. Cross-Cultural Impression Management: A Cultural Knowledge Audit Model

    Science.gov (United States)

    Spong, Abigail; Kamau, Caroline

    2012-01-01

    Purpose: Many people moving into a new culture for work or study do so without prior cross-cultural training, yet successful cultural adaptation has important ramifications. The purpose of this paper is to focus on cross-cultural impression management as an element of cultural adaptation. Does cultural adaptation begin by paying strong attention…

  14. Dynamic global sensitivity analysis in bioreactor networks for bioethanol production.

    Science.gov (United States)

    Ochoa, M P; Estrada, V; Di Maggio, J; Hoch, P M

    2016-01-01

    Dynamic global sensitivity analysis (GSA) was performed for three different dynamic bioreactor models of increasing complexity: a fermenter for bioethanol production, a bioreactors network, where two types of bioreactors were considered: aerobic for biomass production and anaerobic for bioethanol production and a co-fermenter bioreactor, to identify the parameters that most contribute to uncertainty in model outputs. Sobol's method was used to calculate time profiles for sensitivity indices. Numerical results have shown the time-variant influence of uncertain parameters on model variables. Most influential model parameters have been determined. For the model of the bioethanol fermenter, μmax (maximum growth rate) and Ks (half-saturation constant) are the parameters with largest contribution to model variables uncertainty; in the bioreactors network, the most influential parameter is μmax,1 (maximum growth rate in bioreactor 1); whereas λ (glucose-to-total sugars concentration ratio in the feed) is the most influential parameter over all model variables in the co-fermentation bioreactor.

  15. Culture in Transition: A learning model

    DEFF Research Database (Denmark)

    Baca, Susan

    2010-01-01

    of spare capacity, desire, focus, and information. By integrating the element of culture, both in the organizational sense and that pertaining to diversity, otherwise overseen aspects of transition are brought into view, with the potential of reducing stress and increasing job satisfaction. The model......This paper addresses the problem of resistance to attempted changes in organizational culture, particularly those involving diversity, by 1) identifying precisely what is meant by organizational as opposed to societal culture, 2) developing a theoretical model of learning useful in contexts...... of organizational transition, and 3) demonstrating the efficacy of the model by using it to explain empirical research findings. It is argued that learning new cultural currency involves the use of active intelligence to locate and answer relevant questions, and further that this process requires the interplay...

  16. Bioreactors and bioseparation.

    Science.gov (United States)

    Zhang, Siliang; Cao, Xuejun; Chu, Ju; Qian, Jiangchao; Zhuang, Yingping

    2010-01-01

    Along with the rapid development of life science, great attention has been increasingly given to the biotechnological products of cell cultivation technology. In the course of industrialization, bioreactor and bioproduct separation techniques are the two essential technical platforms. In this chapter, the current situation and development prospects of bioreactor techniques in China are systematically discussed, starting with the elucidation of bioreactor processes and the principle of process optimization. Separation technology for biological products is also briefly introduced.At present, a series of bioreactors made by Chinese enterprises have been widely used for laboratory microbial cultivation, process optimization studies, and large-scale production. In the course of bioprocess optimization studies, the complicated bioprocesses in a bioreactor could be resolved into different reaction processes on three scales, namely genetic, cellular, and bioreactor scales. The structural varieties and nonlinear features of various scales of bioprocess systems was discussed through considering the mutual effects of different scale events, namely material flux, energy flux, and information flux, and the optimization approach for bioprocesses was proposed by taking the analysis of metabolic flux and multiscale consideration as a core strategy.In order to realize such an optimization approach, a bioreactor system based on association analysis of multiscale parameters was elaborated, and process optimization of many biological products were materialized, which resulted in great improvement in production efficiency. In designing and manufacturing large-scale bioreactors, the principle of scaling up a process incorporated with flow field study and physiological features in a bioreactor was suggested according to the criterion for the scale-up of cellular physiological and metabolic traits. The flow field features of a bioreactor were investigated through computational fluid

  17. Microbial acetate oxidation in horizontal rotating tubular bioreactor

    Indian Academy of Sciences (India)

    A Slavica; B Šantek; S Novak; V Marić

    2004-06-01

    The aim of this work was to investigate the possibility of conducting a continuous aerobic bioprocess in a horizontal rotating tubular bioreactor (HRTB). Aerobic oxidation of acetate by the action of a mixed microbial culture was chosen as a model process. The microbial culture was not only grown in a suspension but also in the form of a biofilm on the interior surface of HRTB. Efficiency of the bioprocess was monitored by determination of the acetate concentration and chemical oxygen demand (COD). While acetate inlet concentration and feeding rate influenced efficiency of acetate oxidation, the bioreactor rotation speed did not influence the bioprocess dynamics significantly. Gradients of acetate concentration and pH along HRTB were more pronounced at lower feeding rates. Volumetric load of acetate was proved to be the most significant parameter. High volumetric loads (above 2 g acetate l–1 h–1) gave poor acetate oxidation efficiency (only 17 to 50%). When the volumetric load was in the range of 0.60–1.75 g acetate l–1 h–1, acetate oxidation efficiency was 50–75%. At lower volumetric loads (0.14–0.58 g acetate l–1 h–1), complete acetate consumption was achieved. On the basis of the obtained results, it can be concluded that HRTB is suitable for conducting aerobic continuous bioprocesses.

  18. Disposable polymeric cryogel bioreactor matrix for therapeutic protein production.

    Science.gov (United States)

    Jain, Era; Kumar, Ashok

    2013-05-01

    Low cost and high efficiency make disposable bioreactors feasible for small-scale therapeutic development and initial clinical trials. We have developed a cryogel-based disposable bioreactor matrix, which has been used for production of protein therapeutics such as urokinase and monoclonal antibodies (mAbs). The protocol discusses the application of a cryogel bioreactor for mAb production. Cryogels composed of either polyacrylamide (PAAm) coupled to gelatin or semi-interpenetrating PAAm-chitosan are synthesized by free-radical polymerization at -12 °C. Hybridoma cells are immobilized over the cryogel bioreactor and incubated for 48 h. Medium is circulated thereafter at 0.2 ml min(-1) and bioreactors can be run continuously for 60 d. The cryogel-based packed-bed bioreactor can be formulated as a monolith or as beads; it also has an efficiency four times what can be obtained using a tissue-culture flask, a high surface-to-volume ratio and effective nutrient transport. After incubation, the bioreactor setup will take about 60 min using a pre-prepared sterilized cryogel.

  19. Development of a scale down cell culture model using multivariate analysis as a qualification tool.

    Science.gov (United States)

    Tsang, Valerie Liu; Wang, Angela X; Yusuf-Makagiansar, Helena; Ryll, Thomas

    2014-01-01

    In characterizing a cell culture process to support regulatory activities such as process validation and Quality by Design, developing a representative scale down model for design space definition is of great importance. The manufacturing bioreactor should ideally reproduce bench scale performance with respect to all measurable parameters. However, due to intrinsic geometric differences between scales, process performance at manufacturing scale often varies from bench scale performance, typically exhibiting differences in parameters such as cell growth, protein productivity, and/or dissolved carbon dioxide concentration. Here, we describe a case study in which a bench scale cell culture process model is developed to mimic historical manufacturing scale performance for a late stage CHO-based monoclonal antibody program. Using multivariate analysis (MVA) as primary data analysis tool in addition to traditional univariate analysis techniques to identify gaps between scales, process adjustments were implemented at bench scale resulting in an improved scale down cell culture process model. Finally we propose an approach for small scale model qualification including three main aspects: MVA, comparison of key physiological rates, and comparison of product quality attributes.

  20. Modeling the genealogy of a cultural trait.

    Science.gov (United States)

    Aguilar, Elliot; Ghirlanda, Stefano

    2015-05-01

    The mathematical study of genealogies has yielded important insights in population biology, such as the ability to estimate the time to the most recent common ancestor (MRCA) of a sample of genetic sequences or of a group of individuals. Here we introduce a model of cultural genealogies that is a step toward answering similar questions for cultural traits. In our model individuals can inherit from a variable, potentially large number of ancestors, rather than from a fixed, small number of ancestors (one or two) as is typical of genetic evolution. We first show that, given a sample of individuals, a cultural common ancestor does not necessarily exist. We then introduce a related concept: the most recent unique ancestor (MRUA), i.e., the most recent single individual who is the earliest cultural ancestor of the sample. We show that, under neutral evolution, the time to the MRUA can be staggeringly larger than the time to MRCA in a single ancestor model, except when the average number of learning opportunities per individuals is small. Our results point out that the properties of cultural genealogies may be very different from those of genetic genealogies, with potential implications for reconstructing the histories of cultural traits.

  1. Monitoring leachables from single-use bioreactor bags for mammalian cell culture by dispersive liquid-liquid microextraction followed by ultra high performance liquid chromatography quadrupole time of flight mass spectrometry.

    Science.gov (United States)

    Dorival-García, N; Bones, J

    2017-08-25

    A method for the identification of leachables in chemically defined media for CHO cell culture using dispersive liquid-liquid microextraction (DLLME) and UHPLC-MS is described. A Box-Behnken design of experiments (DoE) approach was applied to obtain the optimum extraction conditions of the target analytes. Performance of DLLME as extraction technique was studied by comparison of two commercial chemically defined media for CHO cell culture. General extraction conditions for any group of leachables, regardless of their specific chemical functionalities can be applied and similar optimum conditions were obtained with the two media. Extraction efficiency and matrix effects were determined. The method was validated using matrix-matched standard calibration followed by recovery assays with spiked samples. Finally, cell culture media was incubated in 7 single use bioreactors (SUBs) from different vendors and analysed. TBPP was not detected in any of the samples, whereas DtBP and TBPP-ox were found in all samples, with bDtBPP detected in six SUBs. This method can be used for early identification of non-satisfactory SUB films for cultivation of CHO cell lines for biopharmaceutical production. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Evaluation of the effects of various culture condition on Cr (VI)reduction by Shewanella oneidensis MR-1 in a novel high-throughputmini-bioreactor

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Yinjie J.; Laidlaw, David; Gani, Kishen; Keasling, Jay D.

    2006-03-16

    The growth and Cr(VI) reduction by Shewanella oneidensisMR-1 was examined using a mini-bioreactor system that independentlymonitors and controls pH, dissolved oxygen, and temperature for each ofits 24, 10-mL reactors. Independent monitoring and control of eachreactor in the cassette allows the exploration of a matrix ofenvironmental conditions known to influence S. oneidensis chromiumreduction. S. oneidensis MR-1 grew in minimal medium without amino acidor vitamin supplementation under aerobic conditions but required serineand glycine supplementation under anaerobic conditions. Growth wasinhibited by dissolved oxygen concentrations>80 percent. Lactatetransformation to acetate was enhanced by low concentration of dissolvedoxygen during the logarithmic growth phase. Between 11 and 35oC, thegrowth rate obeyed the Arrhenius reaction rate-temperature relationship,with a maximum growth rate occurring at 35oC. S. oneidensis MR-1 was ableto grow over a wide range of pH (6-9). At neutral pH and temperaturesranging from 30-35oC, S. oneidensis MR-1 reduced 100 mu M Cr(VI) toCr(III) within 20 minutes in the exponential growth phase, and the growthrate was not affected by the addition of chromate; it reduced chromateeven faster at temperatures between 35 and 39oC. At low temperatures(<25oC), acidic (pH<6.5), or alkaline (pH>8.5) conditions, 100mu M Cr(VI) strongly inhibited growth and chromate reduction. Themini-bioreactor system enabled the rapid determination of theseparameters reproducibly and easily by performing very few experiments.Besides its use for examining parameters of interest to environmentalremediation, the device will also allow one to quickly assess parametersfor optimal production of recombinant proteins or secondarymetabolites

  3. What is culture in «cultural economy»? Defining culture to create measurable models in cultural economy

    Directory of Open Access Journals (Sweden)

    Aníbal Monasterio Astobiza

    2017-07-01

    Full Text Available The idea of culture is somewhat vague and ambiguous for the formal goals of economics. The aim of this paper is to define the notion of culture better so as to help build economic explanations based on culture and therefore to measure its impact in every activity or beliefs associated with culture. To define culture according to the canonical evolutionary definition, it is any kind of ritualised behaviour that becomes meaningful for a group and that remains more or less constant and is transmitted down through the generations. Economic institutions are founded, implicitly or explicitly, on a worldview of how humans function; culture is an essential part of understanding us as humans, making it necessary to describe what we understand by culture correctly. In this paper we review the literature on evolutionary anthropology and psychology dealing with the concept of culture to warn that economic modelling ignores intangible benefits of culture rendering economics unable to measure certain cultural items in the digital consumer society.

  4. Thin film bioreactors in space

    Science.gov (United States)

    Hughes-Fulford, M.; Scheld, H. W.

    1989-01-01

    Studies from the Skylab, SL-3 and D-1 missions have demonstrated that biological organisms grown in microgravity have changes in basic cellular functions such as DNA, mRNA and protein synthesis, cytoskeleton synthesis, glucose utilization, and cellular differentiation. Since microgravity could affect prokaryotic and eukaryotic cells at a subcellular and molecular level, space offers an opportunity to learn more about basic biological systems with one inmportant variable removed. The thin film bioreactor will facilitate the handling of fluids in microgravity, under constant temperature and will allow multiple samples of cells to be grown with variable conditions. Studies on cell cultures grown in microgravity would make it possible to identify and quantify changes in basic biological function in microgravity which are needed to develop new applications of orbital research and future biotechnology.

  5. Thin film bioreactors in space

    Science.gov (United States)

    Hughes-Fulford, M.; Scheld, H. W.

    Studies from the Skylab, SL-3 and D-1 missions have demonstrated that biological organisms grown in microgravity have changes in basic cellular functions such as DNA, mRNA and protein synthesis, cytoskeleton synthesis, glucose utilization and cellular differentiation. Since microgravity could affect prokaryotic and eukaryotic cells at a subcellular and molecular level, space offers us an opportunity to learn more about basic biological systems with one important variable removed. The thin film bioreactor will facilitate the handling of fluids in microgravity, under constant temperature and will allow multiple samples of cells to be grown with variable conditions. Studies on cell cultures grown in microgravity would enable us to identify and quantify changes in basic biological function in microgravity which are needed to develop new applications of orbital research and future biotechnology.

  6. Functional study on two artificial liver bioreactors with collagen gel

    Directory of Open Access Journals (Sweden)

    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

  7. Basic bioreactor design.

    NARCIS (Netherlands)

    Riet, van 't K.; Tramper, J.

    1991-01-01

    Based on a graduate course in biochemical engineering, provides the basic knowledge needed for the efficient design of bioreactors and the relevant principles and data for practical process engineering, with an emphasis on enzyme reactors and aerated reactors for microorganisms. Includes exercises.

  8. Basic bioreactor design.

    NARCIS (Netherlands)

    Riet, van 't K.; Tramper, J.

    1991-01-01

    Based on a graduate course in biochemical engineering, provides the basic knowledge needed for the efficient design of bioreactors and the relevant principles and data for practical process engineering, with an emphasis on enzyme reactors and aerated reactors for microorganisms. Includes exercises.

  9. 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 stoichiometry ,ii) processes with a consta

  10. 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

  11. Modelling of a Batch Whey Cultivation of Kluyveromyces marxianus var. lactis MC 5 with Investigation of Mass Transfer Processes in the Bioreactor

    Directory of Open Access Journals (Sweden)

    Mitko Petrov

    2015-04-01

    Full Text Available This study presents a mathematical model of a batch fermentation of lactose oxidation from a natural substratum in a cultivation by the strain Kluyweromyces marxianus var. lactis MC 5. In the model of the process, the mass transfer in the bioreactor for oxygen concentration in the gas phase (GP and in the liquid phase (LP is based on the dispersion model of the GP. In addition, perfect mixing in LP is included. Nine models were investigated for specific growth rate and specific oxygen consumptions rate: Monod, Mink, Tessier, Aiba, Andrews, Haldane, Luong, Edward and Han-Levenspiel. In regard to the parameter estimation, the worst observed error was used for all experiments as an objective function. This approach is a special case of multi objective parameter estimation problems allowing the parameter estimation problem to become a min-max problem. The results obtained (values of criteria, relative error and statistics λ for the specific growth rate showed that the best fit to experimental data is achieved when applying the Mink model. In a combination a Mink, and Monod, Mink, Luong, Haldane, and Han-Levenspiel are used for specific oxygen consumptions rate. Based on the investigation, it was discovered that the best fit belonged to the models of Mink and Haldane, Mink and Luong and Mink and Han-Levenspiel. Therefore, these particular models are used for modeling the batch processes.

  12. Validation of Bioreactor and Human-on-a-Chip Devices for Chemical Safety Assessment.

    Science.gov (United States)

    Rebelo, Sofia P; Dehne, Eva-Maria; Brito, Catarina; Horland, Reyk; Alves, Paula M; Marx, Uwe

    2016-01-01

    Equipment and device qualification and test assay validation in the field of tissue engineered human organs for substance assessment remain formidable tasks with only a few successful examples so far. The hurdles seem to increase with the growing complexity of the biological systems, emulated by the respective models. Controlled single tissue or organ culture in bioreactors improves the organ-specific functions and maintains their phenotypic stability for longer periods of time. The reproducibility attained with bioreactor operations is, per se, an advantage for the validation of safety assessment. Regulatory agencies have gradually altered the validation concept from exhaustive "product" to rigorous and detailed process characterization, valuing reproducibility as a standard for validation. "Human-on-a-chip" technologies applying micro-physiological systems to the in vitro combination of miniaturized human organ equivalents into functional human micro-organisms are nowadays thought to be the most elaborate solution created to date. They target the replacement of the current most complex models-laboratory animals. Therefore, we provide here a road map towards the validation of such "human-on-a-chip" models and qualification of their respective bioreactor and microchip equipment along a path currently used for the respective animal models.

  13. Production of bacterial cellulose membranes in a modified airlift bioreactor by Gluconacetobacter xylinus.

    Science.gov (United States)

    Wu, Sheng-Chi; Li, Meng-Hsun

    2015-10-01

    In this study, a novel bioreactor for producing bacterial cellulose (BC) is proposed. Traditional BC production uses static culture conditions and produces a gelatinous membrane. The potential for using various types of bioreactor, including a stirred tank, conventional airlift, and modified airlift with a rectangular wire-mesh draft tube, in large-scale production has been investigated. The BC obtained from these bioreactors is fibrous or in pellet form. Our proposed airlift bioreactor produces a membrane-type BC from Gluconacetobacter xylinus, the water-holding capacity of which is greater than that of cellulose types produced using static cultivation methods. The Young's modulus of the product can be manipulated by varying the number of net plates in the modified airlift bioreactor. The BC membrane produced using the proposed bioreactor exhibits potential for practical application.

  14. Clinical scale rapid expansion of lymphocytes for adoptive cell transfer therapy in the WAVE® bioreactor

    Science.gov (United States)

    2012-01-01

    Background To simplify clinical scale lymphocyte expansions, we investigated the use of the WAVE®, a closed system bioreactor that utilizes active perfusion to generate high cell numbers in minimal volumes. Methods We have developed an optimized rapid expansion protocol for the WAVE bioreactor that produces clinically relevant numbers of cells for our adoptive cell transfer clinical protocols. Results TIL and genetically modified PBL were rapidly expanded to clinically relevant scales in both static bags and the WAVE bioreactor. Both bioreactors produced comparable numbers of cells; however the cultures generated in the WAVE bioreactor had a higher percentage of CD4+ cells and had a less activated phenotype. Conclusions The WAVE bioreactor simplifies the process of rapidly expanding tumor reactive lymphocytes under GMP conditions, and provides an alternate approach to cell generation for ACT protocols. PMID:22475724

  15. Clinical scale rapid expansion of lymphocytes for adoptive cell transfer therapy in the WAVE® bioreactor

    Directory of Open Access Journals (Sweden)

    Somerville Robert PT

    2012-04-01

    Full Text Available Abstract Background To simplify clinical scale lymphocyte expansions, we investigated the use of the WAVE®, a closed system bioreactor that utilizes active perfusion to generate high cell numbers in minimal volumes. Methods We have developed an optimized rapid expansion protocol for the WAVE bioreactor that produces clinically relevant numbers of cells for our adoptive cell transfer clinical protocols. Results TIL and genetically modified PBL were rapidly expanded to clinically relevant scales in both static bags and the WAVE bioreactor. Both bioreactors produced comparable numbers of cells; however the cultures generated in the WAVE bioreactor had a higher percentage of CD4+ cells and had a less activated phenotype. Conclusions The WAVE bioreactor simplifies the process of rapidly expanding tumor reactive lymphocytes under GMP conditions, and provides an alternate approach to cell generation for ACT protocols.

  16. Cultivation of mammalian cells using a single-use pneumatic bioreactor system.

    Science.gov (United States)

    Obom, Kristina M; Cummings, Patrick J; Ciafardoni, Janelle A; Hashimura, Yasunori; Giroux, Daniel

    2014-10-10

    Recent advances in mammalian, insect, and stem cell cultivation and scale-up have created tremendous opportunities for new therapeutics and personalized medicine innovations. However, translating these advances into therapeutic applications will require in vitro systems that allow for robust, flexible, and cost effective bioreactor systems. There are several bioreactor systems currently utilized in research and commercial settings; however, many of these systems are not optimal for establishing, expanding, and monitoring the growth of different cell types. The culture parameters most challenging to control in these systems include, minimizing hydrodynamic shear, preventing nutrient gradient formation, establishing uniform culture medium aeration, preventing microbial contamination, and monitoring and adjusting culture conditions in real-time. Using a pneumatic single-use bioreactor system, we demonstrate the assembly and operation of this novel bioreactor for mammalian cells grown on micro-carriers. This bioreactor system eliminates many of the challenges associated with currently available systems by minimizing hydrodynamic shear and nutrient gradient formation, and allowing for uniform culture medium aeration. Moreover, the bioreactor's software allows for remote real-time monitoring and adjusting of the bioreactor run parameters. This bioreactor system also has tremendous potential for scale-up of adherent and suspension mammalian cells for production of a variety therapeutic proteins, monoclonal antibodies, stem cells, biosimilars, and vaccines.

  17. Inferring cultural models from corpus data

    DEFF Research Database (Denmark)

    Jensen, Kim Ebensgaard

    2015-01-01

    on three instantiations of the construction – namely, [too young to V], [too proud to V], and [too macho to V] – to see whether there are patterns in their ranges of coattracted verbs that are indicative of force-dynamic relations in cultural models of age, pride, and machismo respectively....

  18. Beyond 3D culture models of cancer

    Science.gov (United States)

    Tanner, Kandice; Gottesman, Michael M.

    2016-01-01

    The mechanisms underlying the spatiotemporal evolution of tumor ecosystems present a challenge in evaluating drug efficacy. In this Perspective, we address the use of three-dimensional in vitro culture models to delineate the dynamic interplay between the tumor and the host microenvironment in an effort to attain realistic platforms for assessing pharmaceutical efficacy in patients. PMID:25877888

  19. Segmented linear modeling of CHO fed‐batch culture and its application to large scale production

    Science.gov (United States)

    Ben Yahia, Bassem; Gourevitch, Boris; Malphettes, Laetitia

    2016-01-01

    ABSTRACT We describe a systematic approach to model CHO metabolism during biopharmaceutical production across a wide range of cell culture conditions. To this end, we applied the metabolic steady state concept. We analyzed and modeled the production rates of metabolites as a function of the specific growth rate. First, the total number of metabolic steady state phases and the location of the breakpoints were determined by recursive partitioning. For this, the smoothed derivative of the metabolic rates with respect to the growth rate were used followed by hierarchical clustering of the obtained partition. We then applied a piecewise regression to the metabolic rates with the previously determined number of phases. This allowed identifying the growth rates at which the cells underwent a metabolic shift. The resulting model with piecewise linear relationships between metabolic rates and the growth rate did well describe cellular metabolism in the fed‐batch cultures. Using the model structure and parameter values from a small‐scale cell culture (2 L) training dataset, it was possible to predict metabolic rates of new fed‐batch cultures just using the experimental specific growth rates. Such prediction was successful both at the laboratory scale with 2 L bioreactors but also at the production scale of 2000 L. This type of modeling provides a flexible framework to set a solid foundation for metabolic flux analysis and mechanistic type of modeling. Biotechnol. Bioeng. 2017;114: 785–797. © 2016 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc. PMID:27869296

  20. Segmented linear modeling of CHO fed-batch culture and its application to large scale production.

    Science.gov (United States)

    Ben Yahia, Bassem; Gourevitch, Boris; Malphettes, Laetitia; Heinzle, Elmar

    2017-04-01

    We describe a systematic approach to model CHO metabolism during biopharmaceutical production across a wide range of cell culture conditions. To this end, we applied the metabolic steady state concept. We analyzed and modeled the production rates of metabolites as a function of the specific growth rate. First, the total number of metabolic steady state phases and the location of the breakpoints were determined by recursive partitioning. For this, the smoothed derivative of the metabolic rates with respect to the growth rate were used followed by hierarchical clustering of the obtained partition. We then applied a piecewise regression to the metabolic rates with the previously determined number of phases. This allowed identifying the growth rates at which the cells underwent a metabolic shift. The resulting model with piecewise linear relationships between metabolic rates and the growth rate did well describe cellular metabolism in the fed-batch cultures. Using the model structure and parameter values from a small-scale cell culture (2 L) training dataset, it was possible to predict metabolic rates of new fed-batch cultures just using the experimental specific growth rates. Such prediction was successful both at the laboratory scale with 2 L bioreactors but also at the production scale of 2000 L. This type of modeling provides a flexible framework to set a solid foundation for metabolic flux analysis and mechanistic type of modeling. Biotechnol. Bioeng. 2017;114: 785-797. © 2016 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc. © 2016 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

  1. Oxygen transfer coefficient and the kinetic parameters of exo-polygalacturonase production by Aspergillus flavipes FP-500 in shake flasks and bioreactor.

    Science.gov (United States)

    Gómez Sánchez, C E; Martínez-Trujillo, A; Aguilar Osorio, G

    2012-12-01

    To evaluate and compare the sensitivity of Exo-PG production and kinetic parameters of Aspergillus flavipes FP-500 to oxygen transfer condition in shake flasks and bioreactor. Aspergillus flavipes FP-500 was grown on pectin as carbon source in shake flasks and bioreactor at different oxygen transfer conditions. The volumetric coefficient of oxygen transfer (kLa) was modified by changing both, the flask size/medium volume ratio and the agitation speed. Higher biomass concentration, Exo-PG activity, maximum specific growth rate and yield coefficient were obtained in bioreactor at higher kLa value. A strong correlation was found between biomass, Exo-PG activity and growth-associated product coefficient to kLa in bioreactor but does not in shake flasks. The mathematical model provided a good description of growth, pectin consumption and Exo-PG production in submerged batch cultures carried out in bioreactor. Biomass concentration, Exo-PG activity and their kinetics of Aspergillus flavipes FP-500 were strongly influenced by oxygen transfer condition and cultivation system. Significance and Impact of Study The production of enzymes by fungal fermentation is strictly aerobic and understanding the influence of oxygen transfer condition on the production kinetic is of vital importance in order to design, optimize and translate bioprocesses to industrial scale. © 2012 The Society for Applied Microbiology.

  2. LTCC based bioreactors for cell cultivation

    Science.gov (United States)

    Bartsch, H.; Welker, T.; Welker, K.; Witte, H.; Müller, J.

    2016-01-01

    LTCC multilayers offer a wide range of structural options and flexibility of connections not available in standard thin film technology. Therefore they are considered as material base for cell culture reactors. The integration of microfluidic handling systems and features for optical and electrical capturing of indicators for cell culture growth offers the platform for an open system concept. The present paper assesses different approaches for the creation of microfluidic channels in LTCC multilayers. Basic functions required for the fluid management in bioreactors include temperature and flow control. Both features can be realized with integrated heaters and temperature sensors in LTCC multilayers. Technological conditions for the integration of such elements into bioreactors are analysed. The temperature regulation for the system makes use of NTC thermistor sensors which serve as real value input for the control of the heater. It allows the adjustment of the fluid temperature with an accuracy of 0.2 K. The tempered fluid flows through the cell culture chamber. Inside of this chamber a thick film electrode array monitors the impedance as an indicator for the growth process of 3-dimensional cell cultures. At the system output a flow sensor is arranged to monitor the continual flow. For this purpose a calorimetric sensor is implemented, and its crucial design parameters are discussed. Thus, the work presented gives an overview on the current status of LTCC based fluid management for cell culture reactors, which provides a promising base for the automation of cell culture processes.

  3. Catalytic bioreactors and methods of using same

    Energy Technology Data Exchange (ETDEWEB)

    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.

  4. Hydrofocusing Bioreactor Produces Anti-Cancer Alkaloids

    Science.gov (United States)

    Gonda, Steve R.; Valluri, Jagan V.

    2011-01-01

    A methodology for growing three-dimensional plant tissue models in a hydrodynamic focusing bioreactor (HFB) has been developed. The methodology is expected to be widely applicable, both on Earth and in outer space, as a means of growing plant cells and aggregates thereof under controlled conditions for diverse purposes, including research on effects of gravitation and other environmental factors upon plant growth and utilization of plant tissue cultures to produce drugs in quantities greater and at costs lower than those of conventional methodologies. The HFB was described in Hydro focus - ing Bioreactor for Three-Dimensional Cell Culture (MSC-22358), NASA Tech Briefs, Vol. 27, No. 3 (March 2003), page 66. To recapitulate: The HFB offers a unique hydrofocusing capability that enables the creation of a low-shear liquid culture environment simultaneously with the herding of suspended cells and tissue assemblies and removal of unwanted air bubbles. The HFB includes a rotating cell-culture vessel with a centrally located sampling port and an internal rotating viscous spinner attached to a rotating base. The vessel and viscous spinner can be made to rotate at the same speed and direction or different speeds and directions to tailor the flow field and the associated hydrodynamic forces in the vessel in order to obtain low-shear suspension of cells and control of the locations of cells and air bubbles. For research and pharmaceutical-production applications, the HFB offers two major benefits: low shear stress, which promotes the assembly of cells into tissue-like three-dimensional constructs; and randomization of gravitational vectors relative to cells, which affects production of medicinal compounds. Presumably, apposition of plant cells in the absence of shear forces promotes cell-cell contacts, cell aggregation, and cell differentiation. Only gentle mixing is necessary for distributing nutrients and oxygen. It has been postulated that inasmuch as cells in the simulated

  5. NASA Bioreactors Advance Disease Treatments

    Science.gov (United States)

    2009-01-01

    The International Space Station (ISS) is falling. This is no threat to the astronauts onboard, however, because falling is part of the ISS staying in orbit. The absence of gravity beyond the Earth s atmosphere is actually an illusion; at the ISS s orbital altitude of approximately 250 miles above the surface, the planet s gravitational pull is only 12-percent weaker than on the ground. Gravity is constantly pulling the ISS back to Earth, but the space station is also constantly traveling at nearly 18,000 miles per hour. This means that, even though the ISS is falling toward Earth, it is moving sideways fast enough to continually miss impacting the planet. The balance between the force of gravity and the ISS s motion creates a stable orbit, and the fact that the ISS and everything in it including the astronauts are falling at an equal rate creates the condition of weightlessness called microgravity. The constant falling of objects in orbit is not only an important principle in space, but it is also a key element of a revolutionary NASA technology here on Earth that may soon help cure medical ailments from heart disease to diabetes. In the mid-1980s, NASA researchers at Johnson Space Center were investigating the effects of long-term microgravity on human tissues. At the time, the Agency s shuttle fleet was grounded following the 1986 Space Shuttle Challenger disaster, and researchers had no access to the microgravity conditions of space. To provide a method for recreating such conditions on Earth, Johnson s David Wolf, Tinh Trinh, and Ray Schwarz developed that same year a horizontal, rotating device called a rotating wall bioreactor that allowed the growth of human cells in simulated weightlessness. Previously, cell cultures on Earth could only be grown two-dimensionally in Petri dishes, because gravity would cause the multiplying cells to sink within their growth medium. These cells do not look or function like real human cells, which grow three-dimensionally in

  6. Characterization of the dynamic behavior of nonlinear biosystems in the presence of model uncertainty using singular invariance PDEs: application to immobilized enzyme and cell bioreactors.

    Science.gov (United States)

    Kazantzis, Nikolaos; Kazantzi, Vasiliki

    2010-04-01

    A new approach to the problem of characterizing the dynamic behavior of nonlinear biosystems in the presence of model uncertainty using the notion of slow invariant manifold is proposed. The problem of interest is addressed within the context of singular partial differential equations (PDE) theory, and in particular, through a system of singular quasi-linear invariance PDEs for which a general set of conditions for solvability is provided. Within the class of analytic solutions, this set of conditions guarantees the existence and uniqueness of a locally analytic solution which represents the system's slow invariant manifold exponentially attracting all dynamic trajectories in the absence of model uncertainty. An exact reduced-order model is then obtained through the restriction of the original biosystem dynamics on the slow manifold. The analyticity property of the solution to the invariance PDEs enables the development of a series solution method that can be easily implemented using MAPLE leading to polynomial approximations up to the desired degree of accuracy. Furthermore, the aforementioned attractivity property and the transition towards the above manifold is analyzed and characterized in the presence of model uncertainty. Finally, examples of certain immobilized enzyme bioreactors are considered to elucidate aspects of the proposed context of analysis.

  7. Scale-up of osmotic membrane bioreactors by modeling salt accumulation and draw solution dilution using hollow-fiber membrane characteristics and operation conditions.

    Science.gov (United States)

    Kim, Suhan

    2014-08-01

    A full-scale osmotic membrane bioreactor (OMBR) model was developed to simulate salt accumulation, draw solution (DS) dilution, and water flux over the hollow-fiber membrane length. The model uses the OMBR design parameters, DS properties, and forward osmosis (FO) membrane characteristics obtained from lab-scale tests. The modeling results revealed a tremendous water flux decline (10→0.82LMH) and short solids retention time (SRT: 5days) due to salt accumulation and DS dilution when OMBR is scaled up using commercially available DS and FO membrane. Simulated water flux is a result of interplay among reverse salt flux, internal and external concentration polarization (ICP and ECP). ECP adversely impacts water flux considerably in full-scale OMBR although it is often ignored in previous works. The OMBR model makes it possible to select better DS properties (higher flow rate and salt concentration) and FO membranes with higher water flux propensity in full-scale operation. Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. 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\\

  9. Cultural competence models and cultural competence assessment instruments in nursing: a literature review.

    Science.gov (United States)

    Shen, Zuwang

    2015-05-01

    The author reviewed cultural competence models and cultural competence assessment instruments developed and published by nurse researchers since 1982. Both models and instruments were examined in terms of their components, theoretical backgrounds, empirical validation, and psychometric evaluation. Most models were not empirically tested; only a few models developed model-based instruments. About half of the instruments were tested with varying levels of psychometric properties. Other related issues were discussed, including the definition of cultural competence and its significance in model and instrument development, limitations of existing models and instruments, impact of cultural competence on health disparities, and further work in cultural competence research and practice. © The Author(s) 2014.

  10. 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

  11. 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.

  12. 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.

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

    Science.gov (United States)

    Barzegari, Abolfazl; Saei, Amir Ata

    2012-01-01

    Introduction The severe need for constructing replacement tissues in organ transplanta-tion 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 chon-drocytes, 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 vari-ous 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. PMID:23678438

  14. 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.

  15. Comparisons of optically monitored small-scale stirred tank vessels to optically controlled disposable bag bioreactors

    Directory of Open Access Journals (Sweden)

    Brorson Kurt A

    2009-08-01

    Full Text Available Abstract Background Upstream bioprocesses are extremely complex since living organisms are used to generate active pharmaceutical ingredients (APIs. Cells in culture behave uniquely in response to their environment, thus culture conditions must be precisely defined and controlled in order for productivity and product quality to be reproducible. Thus, development culturing platforms are needed where many experiments can be carried out at once and pertinent scale-up information can be obtained. Results Here we have tested a High Throughput Bioreactor (HTBR as a scale-down model for a lab-scale wave-type bioreactor (CultiBag. Mass transfer was characterized in both systems and scaling based on volumetric oxygen mass transfer coefficient (kLa was sufficient to give similar DO trends. HTBR and CultiBag cell growth and mAb production were highly comparable in the first experiment where DO and pH were allowed to vary freely. In the second experiment, growth and mAb production rates were lower in the HTBR as compared to the CultiBag, where pH was controlled. The differences in magnitude were not considered significant for biological systems. Conclusion Similar oxygen delivery rates were achieved in both systems, leading to comparable culture performance (growth and mAb production across scales and mode of mixing. HTBR model was most fitting when neither system was pH-controlled, providing an information-rich alternative to typically non-monitored mL-scale platforms.

  16. Microbial abatement of toluene using Aspergillus niger in upflow bioreactor.

    Science.gov (United States)

    Gopinath, M; Mohanapriya, C; Sivakumar, K; Baskar, G; Muthukumaran, C; Dhanasekar, R

    2016-12-01

    Microbial abatement of toluene using Aspergillus niger in coir packed upflow bioreactor was investigated in this study. Toluene degrading microbes were isolated from municipal sewage effluent and identified by 16s rRNA sequencing method. The microbes were cultured in 2% (v/v) toluene input per day, which exhibited 95% removal efficiency with the kinetic correction value (R(2)) of 0.9024 at the optimum flow rate of about 0.4m(3)h(-1). Various parameters such as effect of flow rate, column height, elimination capacity and EBRT with removal efficiency for 50 day cycle were also optimized. The plug flow model for toluene degradation was properly expressed and the Monod kinetics constant Km and rmax values were determined as 2.25gm(-3) and 67.773gm(-3)h(-1) respectively for microbial growth rate. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. 光照对光生物反应器中微藻高密度光自养培养的影响%Effect of Illumination on Microalgae Cultured at High Cell Density in Photo-bioreactor

    Institute of Scientific and Technical Information of China (English)

    李永富; 孟范平; 李祥蕾; 马冬冬

    2013-01-01

    光生物反应器是实现微藻高密度培养的重要装置,其设计的关键技术之一是选择合适的光照方式.根据国内外近十年来的相关研究成果,重点介绍了入射光性质(光源、光强、光质和光暗循环)和光能分布对微藻生长的影响,评述了用于微藻高密度培养的光照技术,展望了进一步的研究方向,为高效光生物反应器的设计和优化提供参考.%Photo-bioreactors (PBR) are important device for realizing high-density culture of microalgae. One of the key technologies for designing PBR is to choose the appropriate illumination mode. According to related research achievements domestic and international in recent years, the present research conditions of illumination technique in PBR were reviewed. An introduction is emphatically given of the effect of two factors as incident light properties (including light source, light intensity, light quality and light dark cycle etc. ) and light energy distribution, on the microalgae growth. The future research directions are also put forward. It could provide reference for the efficient PBR system design and optimize.

  18. Bioprocess kinetics in a horizontal rotating tubular bioreactor.

    Science.gov (United States)

    Ivancić, M; Santek, B; Novak, S; Horvat, P; Marić, V

    2004-04-01

    A horizontal rotating tubular bioreactor (HRTB) is a plug flow bioreactor whose interior is provided with O-ring-shaped partition walls that serve as carriers for microbial biomass. During this investigation, microbial biomass was grown in suspension and on the bioreactor inner surface as a microbial biofilm with average mass that was considerably higher than suspended biomass. The dynamics of bioprocess in HRTB was studied by different combinations of process parameters (bioreactor rotation speed and mean residence time) and it was monitored by withdrawing the samples from five positions along the bioreactor. During this investigation it was also observed that mean residence time had a more pronounced effect on the bioprocess dynamics than bioreactor rotation speed. For the description of bioprocess kinetics in HRTB an unstructured kinetic model was established that defines biomass growth, product formations and substrate consumption rate by using a modified Monod (Levenspiel) model. This kinetic model defines changes in suspension and in microbial biofilm, and it shows relatively good agreement with experimental data.

  19. Imaging glucose metabolism in perfluorocarbon-perfused hepatocyte bioreactors using positron emission tomography.

    Science.gov (United States)

    Nieuwoudt, Martin; Wiggett, Scholtz; Malfeld, Susan; van der Merwe, Schalk W

    2009-01-01

    In vitro hepatocyte bioreactor functionality depends particularly on maintaining appropriate oxygen levels and exposure to nonparenchymal cells. An attractive solution without immunological consequences to the patient is incorporating a perfluorocarbon oxygen carrier in the circulating medium and co-culturing hepatocytes with stellate cells. Since bioreactors are normally sealed sterile units, demonstrating metabolic functionality is hindered by limited access to the cells after their aggregation in the matrix. A novel possibility is to use positron emission tomography (PET) to image cellular radioactive glucose uptake under O(2)-limited conditions. In this study, primary cell isolation procedures were carried out on eight pigs. Pairs of cell-seeded and cell-free (control) bioreactors with and without perfluorocarbon were cultured under identical conditions and were oxygenated using hypoxic (5% O(2)) and ambient (20% O(2)) gas mixes. Sixteen PET scans were conducted 24 h after cell isolation, the same timescale as that involved in treating a liver failure patient with a primary-cell bioreactor. In all cases, cell-seeded bioreactors without perfluorocarbon were more radioactive, i.e., were more glycolytic, than those with perfluorocarbon. This difference was significant in the hypoxic pair of bioreactors but not in the ambient pair of bioreactors. Additionally, in the same hypoxic bioreactors, circulating extracellular steady-state glucose levels were significantly lower and lactate levels were higher than those in the ambient bioreactors. Similar findings have been made in other in vitro hepatocyte studies investigating the effects of perfluorocarbons. PET is attractive for studying in situ O(2)-dependent bioreactor metabolism because of its visual and numerically quantifiable outputs. Longer-term metabolic studies (e.g., 5-10 days) investigating the effect of perfluorocarbon on bioreactor longevity will complement these findings in the future.

  20. A comparative study of the chondrogenic potential between synthetic and natural scaffolds in an in vivo bioreactor

    Science.gov (United States)

    Huang, Jung-Ju; Yang, Shu-Rui; Chu, I.-Ming; Brey, Eric M.; Hsiao, Hui-Yi; Cheng, Ming-Huei

    2013-10-01

    The clinical demand for cartilage tissue engineering is potentially large for reconstruction defects resulting from congenital deformities or degenerative disease due to limited donor sites for autologous tissue and donor site morbidities. Cartilage tissue engineering has been successfully applied to the medical field: a scaffold pre-cultured with chondrocytes was used prior to implantation in an animal model. We have developed a surgical approach in which tissues are engineered by implantation with a vascular pedicle as an in vivo bioreactor in bone and adipose tissue engineering. Collagen type II, chitosan, poly(lactic-co-glycolic acid) (PLGA) and polycaprolactone (PCL) were four commonly applied scaffolds in cartilage tissue engineering. To expand the application of the same animal model in cartilage tissue engineering, these four scaffolds were selected and compared for their ability to generate cartilage with chondrocytes in the same model with an in vivo bioreactor. Gene expression and immunohistochemistry staining methods were used to evaluate the chondrogenesis and osteogenesis of specimens. The result showed that the PLGA and PCL scaffolds exhibited better chondrogenesis than chitosan and type II collagen in the in vivo bioreactor. Among these four scaffolds, the PCL scaffold presented the most significant result of chondrogenesis embedded around the vascular pedicle in the long-term culture incubation phase.

  1. Aujeszky's disease virus production in disposable bioreactor

    Indian Academy of Sciences (India)

    I Slivac; V Gaurina Srček; K Radošević; I Kmetič; Z Kniewald

    2006-09-01

    A novel, disposable-bag bioreactor system that uses wave action for mixing and transferring oxygen was evaluated for BHK 21 C13 cell line growth and Aujeszky’s disease virus (ADV) production. Growth kinetics of BHK 21 C13 cells in the wave bioreactor during 3-day period were determined. At the end of the 3-day culture period and cell density of 1.82 × 106 cells ml–1, the reactor was inoculated with 9 ml of gE- Bartha K-61 strain ADV suspension (105.9 TCID50) with multiplicity of infection (MOI) of 0.01. After a 144 h incubation period, 400 ml of ADV harvest was obtained with titre of 107.0 TCID50 ml–1, which corresponds to 40,000 doses of vaccine against AD. In conclusion, the results obtained with the wave bioreactor using BHK 21 C13 cells showed that this system can be considered as suitable for ADV or BHK 21 C13 cell biomass production.

  2. Oxygen transfer in a pressurized airlift bioreactor.

    Science.gov (United States)

    Campani, Gilson; Ribeiro, Marcelo Perencin Arruda; Horta, Antônio Carlos Luperni; Giordano, Roberto Campos; Badino, Alberto Colli; Zangirolami, Teresa Cristina

    2015-08-01

    Airlift bioreactors (ALBs) offer advantages over conventional systems, such as simplicity of construction, reduced risk of contamination, and efficient gas-liquid dispersion with low power consumption. ALBs are usually operated under atmospheric pressure. However, in bioprocesses with high oxygen demand, such as high cell density cultures, oxygen limitation may occur even when operating with high superficial gas velocity and air enriched with oxygen. One way of overcoming this drawback is to pressurize the reactor. In this configuration, it is important to assess the influence of bioreactor internal pressure on the gas hold-up, volumetric oxygen transfer coefficient (k(L)a), and volumetric oxygen transfer rate (OTR). Experiments were carried out in a concentric-tube airlift bioreactor with a 5 dm(3) working volume, equipped with a system for automatic monitoring and control of the pressure, temperature, and inlet gas flow rate. The results showed that, in disagreement with previous published results for bubble column and external loop airlift reactors, overpressure did not significantly affect k(L)a within the studied ranges of pressure (0.1-0.4 MPa) and superficial gas velocity in the riser (0.032-0.065 m s(-1)). Nevertheless, a positive effect on OTR was observed: it increased up to 5.4 times, surpassing by 2.3 times the oxygen transfer in a 4 dm(3) stirred tank reactor operated under standard cultivation conditions. These results contribute to the development of non-conventional reactors, especially pneumatic bioreactors operated using novel strategies for oxygen control.

  3. Tissue engineering bioreactor systems for applying physical and electrical stimulations to cells.

    Science.gov (United States)

    Jin, GyuHyun; Yang, Gi-Hoon; Kim, GeunHyung

    2015-05-01

    Bioreactor systems in tissue engineering applications provide various types of stimulation to mimic the tissues in vitro and in vivo. Various bioreactors have been designed to induce high cellular activities, including initial cell attachment, cell growth, and differentiation. Although cell-stimulation processes exert mostly positive effects on cellular responses, in some cases such stimulation can also have a negative effect on cultured cells. In this review, we discuss various types of bioreactor and the positive and negative effects of stimulation (physical, chemical, and electrical) on various cultured cell types.

  4. Progress toward forecasting product quality and quantity of mammalian cell culture processes by performance-based modeling.

    Science.gov (United States)

    Schmidberger, Timo; Posch, Christoph; Sasse, Alexandra; Gülch, Carina; Huber, Robert

    2015-01-01

    The production of biopharmaceuticals requires highly sophisticated, complex cell based processes. Once a process has been developed, acceptable ranges for various control parameters are typically defined based on process characterization studies often comprising several dozens of small scale bioreactor cultivations. A lot of data is generated during these studies and usually only the information needed to define acceptable ranges is processed in more detail. Making use of the wealth of information contained in such data sets, we present here a methodology that uses performance data (such as metabolite profiles) to forecast the product quality and quantity of mammalian cell culture processes based on a toolbox of advanced statistical methods. With this performance based modeling (PBM) the final product concentration and 12 quality attributes (QAs) for two different biopharmaceutical products were predicted in daily intervals throughout the main stage process. The best forecast was achieved for product concentration in a very early phase of the process. Furthermore, some glycan isoforms were predicted with good accuracy several days before the bioreactor was harvested. Overall, PBM clearly demonstrated its capability of early process endpoint prediction by only using commonly available data, even though it was not possible to predict all QAs with the desired accuracy. Knowing the product quality prior to the harvest allows the manufacturer to take counter measures in case the forecasted quality or quantity deviates from what is expected. This would be a big step towards real-time release, an important element of the FDA's PAT initiative. © 2015 American Institute of Chemical Engineers.

  5. Mathematical model analysis on the enhancement of aeration efficiency using ladder-type flat membrane module forms in the Submerged Membrane Bio-reactor (SMBR)

    Institute of Scientific and Technical Information of China (English)

    LI Bo; YE MaoSheng; YANG FengLin; MA Hui

    2009-01-01

    The cross-flow shearing action produced from the inferior aeration in the Submerged Membrane Bio-reactor (SMBR) Is an effective way to further improve anti-fouling effects of membrane modules.Based on the widely-applied vertical structure of flat membrane modules, improvements are made that ladder-type flat membrane structure is designed with a certain inclined angle θ so that the cross-flow velocity of bubble near the membrane surface can be held, and the intensity and times of elastic colli-sion between bubbles and membrane surface can be increased. This can improve scouring action ofmembrane surface on aeration and reduce energy consumption of strong aeration in SMBR. By de-ducing and improving the mathematics model of collision between bubble and vertical flat put forward by Vries, the relatively suitable Incline angle θ under certain aeration place and in certain size rang ofbubble can be obtained with the computer iterative calculation technology. Finally, for many groups of ladder-type flat membrane in parallel placement in the practical application of SMBR, some sugges-tions are offered: the interval distance of membrane modules is 8--15 mm, and aeration should be op-erated at 5--7 mm among membrane modules, and the optimal design angle of trapeziform membrane is 1.7°--2.5°.

  6. RWPV bioreactor mass transport: earth-based and in microgravity

    Science.gov (United States)

    Begley, Cynthia M.; Kleis, Stanley J.

    2002-01-01

    Mass transport and mixing of perfused scalar quantities in the NASA Rotating Wall Perfused Vessel bioreactor are studied using numerical models of the flow field and scalar concentration field. Operating conditions typical of both microgravity and ground-based cell cultures are studied to determine the expected vessel performance for both flight and ground-based control experiments. Results are presented for the transport of oxygen with cell densities and consumption rates typical of colon cancer cells cultured in the RWPV. The transport and mixing characteristics are first investigated with a step change in the perfusion inlet concentration by computing the time histories of the time to exceed 10% inlet concentration. The effects of a uniform cell utilization rate are then investigated with time histories of the outlet concentration, volume average concentration, and volume fraction starved. It is found that the operating conditions used in microgravity produce results that are quite different then those for ground-based conditions. Mixing times for microgravity conditions are significantly shorter than those for ground-based operation. Increasing the differential rotation rates (microgravity) increases the mixing and transport, while increasing the mean rotation rate (ground-based) suppresses both. Increasing perfusion rates enhances mass transport for both microgravity and ground-based cases, however, for the present range of operating conditions, above 5-10 cc/min there are diminishing returns as much of the inlet fluid is transported directly to the perfusion exit. The results show that exit concentration is not a good indicator of the concentration distributions in the vessel. In microgravity conditions, the NASA RWPV bioreactor with the viscous pump has been shown to provide an environment that is well mixed. Even when operated near the theoretical minimum perfusion rates, only a small fraction of the volume provides less than the required oxygen levels

  7. Effects of a perfusion bioreactor activated novel bone substitute in spine fusion in sheep

    DEFF Research Database (Denmark)

    Sørensen, Jesper Roed; Koroma, Kariatta Ester; Ding, Ming

    2012-01-01

    To evaluate the effect of a large perfusion-bioreactor cell-activated bone substitute, on a two-level large posterolateral spine fusion sheep model.......To evaluate the effect of a large perfusion-bioreactor cell-activated bone substitute, on a two-level large posterolateral spine fusion sheep model....

  8. Analysis of organizational culture with social network models

    OpenAIRE

    Titov, S.

    2015-01-01

    Organizational culture is nowadays an object of numerous scientific papers. However, only marginal part of existing research attempts to use the formal models of organizational cultures. The lack of organizational culture models significantly limits the further research in this area and restricts the application of the theory to practice of organizational culture change projects. The article consists of general views on potential application of network models and social network analysis to th...

  9. Effect of pH Heterogeneity in Large-scale Bioreactor on Fed-batch Culture Process of CHO cells%大型反应器内pH不均一性对CHO细胞流加培养过程的影响

    Institute of Scientific and Technical Information of China (English)

    刘金涛; 王星懿; 范里; 邓献存; 刘旭平; 谭文松

    2015-01-01

    In order to study the effect of pH heterogeneity in large-scale bioreactor on cell culture process of CHO cells, we established a scale down model consistent of stirred tank reactor and plug flow reactor to simulate the pH heterogeneity of large-scale bioreactor based on the mixing characteristic. The results showed that the scale down process with 30 s residence time has no statically difference with the control process. However, significant effect on cell growth, cell metabolism and protein production were found when increased the residence time of PFR. Cell growth rate decreased accompanied by tremendously increase of ammonia and lactate when increased the pH heterogeneity. In addition, the titer, sialic acid content and bioactivity of antibody fusion protein were also decreased when increased the pH heterogeneity.%为了研究大型反应器中pH不均一性对CHO细胞流加培养过程的影响,并将培养过程顺利地放大到生产规模,根据大型反应器的混合特性,构建了搅拌式反应器与平推流反应器串联的规模缩小装置用于模拟大型反应器中的pH不均一性。结果表明停留时间为30 s时,整个培养过程和对照相比并无显著的差异,这表明此时补碱所导致的pH不均一性并未对流加培养工艺造成影响。而随着停留时间的延长,反应器内pH不均一的程度越大,细胞生长和产物表达受到抑制越明显;与此同时,乳酸和氨的累积显著增加,而关键质量属性唾液酸和生物学活性也随之降低。

  10. Regulation of mesenchymal stem cell 3D microenvironment: From macro to microfluidic bioreactors.

    Science.gov (United States)

    Sart, Sébastien; Agathos, Spiros N; Li, Yan; Ma, Teng

    2016-01-01

    Human mesenchymal stem cells (hMSCs) have emerged as an important cell type in cell therapy and tissue engineering. In these applications, maintaining the therapeutic properties of hMSCs requires tight control of the culture environments and the structural cell organizations. Bioreactor systems are essential tools to achieve these goals in the clinical-scale expansion and tissue engineering applications. This review summarizes how different bioreactors provide cues to regulate the structure and the chemico-mechanical microenvironment of hMSCs with a focus on 3D organization. In addition to conventional bioreactors, recent advances in microfluidic bioreactors as a novel approach to better control the hMSC microenvironment are also discussed. These advancements highlight the key role of bioreactor systems in preserving hMSC's functional properties by providing dynamic and temporal regulation of in vitro cellular microenvironment.

  11. Production of Calcaride A by Calcarisporium sp. in Shaken Flasks and Stirred Bioreactors.

    Science.gov (United States)

    Tamminen, Anu; Wang, Yanming; Wiebe, Marilyn G

    2015-06-24

    Increased interest in marine resources has led to increased screening of marine fungi for novel bioactive compounds and considerable effort is being invested in discovering these metabolites. For compound discovery, small-scale cultures are adequate, but agitated bioreactors are desirable for larger-scale production. Calcarisporium sp. KF525 has recently been described to produce calcaride A, a cyclic polyester with antibiotic activity, in agitated flasks. Here, we describe improvements in the production of calcaride A in both flasks (13-fold improvement) and stirred bioreactors (200-fold improvement). Production of calcaride A in bioreactors was initially substantially lower than in shaken flasks. The cultivation pH (reduced from 6.8 to bioreactor, which was only slightly less than in shaken flasks (14 mg·g-1, 100 mg·L-1). The results demonstrate that a scalable process for calcaride A production could be developed using an iterative approach with flasks and bioreactors.

  12. Coupling of mixing and microbial kinetics for evaluating the performance of bioreactors

    Energy Technology Data Exchange (ETDEWEB)

    Bajpai, R.K.; Reuss, M.

    1982-06-01

    The effect of the dynamics of the mixing process in mechanically stirred bioreactors upon their performance has been investigated by using a circulation-model for flow of fluid and a two-environment- model to account for the extent of micromixing in the vessel. With the help of an empirical kinetic model to depict the oxidative and the fermentative growth of bakers' yeast, experimentally observed effects of initial sugar concentrations upon critical dilution rates in continuous cultures of this system could be explained. Effect of different scales of operation upon productivities of fed-batch cultures of bakers' yeast could also be satisfactory predicted. A discrete simulation procedure was found to be a convenient tool in coupling the two-environment circulation-model of mixing with kinetics of microbial reactions. (Refs. 31).

  13. Integrating human stem cell expansion and neuronal differentiation in bioreactors

    Directory of Open Access Journals (Sweden)

    Costa Eunice M

    2009-09-01

    Full Text Available Abstract Background Human stem cells are cellular resources with outstanding potential for cell therapy. However, for the fulfillment of this application, major challenges remain to be met. Of paramount importance is the development of robust systems for in vitro stem cell expansion and differentiation. In this work, we successfully developed an efficient scalable bioprocess for the fast production of human neurons. Results The expansion of undifferentiated human embryonal carcinoma stem cells (NTera2/cl.D1 cell line as 3D-aggregates was firstly optimized in spinner vessel. The media exchange operation mode with an inoculum concentration of 4 × 105 cell/mL was the most efficient strategy tested, with a 4.6-fold increase in cell concentration achieved in 5 days. These results were validated in a bioreactor where similar profile and metabolic performance were obtained. Furthermore, characterization of the expanded population by immunofluorescence microscopy and flow cytometry showed that NT2 cells maintained their stem cell characteristics along the bioreactor culture time. Finally, the neuronal differentiation step was integrated in the bioreactor process, by addition of retinoic acid when cells were in the middle of the exponential phase. Neurosphere composition was monitored and neuronal differentiation efficiency evaluated along the culture time. The results show that, for bioreactor cultures, we were able to increase significantly the neuronal differentiation efficiency by 10-fold while reducing drastically, by 30%, the time required for the differentiation process. Conclusion The culture systems developed herein are robust and represent one-step-forward towards the development of integrated bioprocesses, bridging stem cell expansion and differentiation in fully controlled bioreactors.

  14. 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.

  15. Modelling the evolution and diversity of cumulative culture

    Science.gov (United States)

    Enquist, Magnus; Ghirlanda, Stefano; Eriksson, Kimmo

    2011-01-01

    Previous work on mathematical models of cultural evolution has mainly focused on the diffusion of simple cultural elements. However, a characteristic feature of human cultural evolution is the seemingly limitless appearance of new and increasingly complex cultural elements. Here, we develop a general modelling framework to study such cumulative processes, in which we assume that the appearance and disappearance of cultural elements are stochastic events that depend on the current state of culture. Five scenarios are explored: evolution of independent cultural elements, stepwise modification of elements, differentiation or combination of elements and systems of cultural elements. As one application of our framework, we study the evolution of cultural diversity (in time as well as between groups). PMID:21199845

  16. Modelling the evolution and diversity of cumulative culture.

    Science.gov (United States)

    Enquist, Magnus; Ghirlanda, Stefano; Eriksson, Kimmo

    2011-02-12

    Previous work on mathematical models of cultural evolution has mainly focused on the diffusion of simple cultural elements. However, a characteristic feature of human cultural evolution is the seemingly limitless appearance of new and increasingly complex cultural elements. Here, we develop a general modelling framework to study such cumulative processes, in which we assume that the appearance and disappearance of cultural elements are stochastic events that depend on the current state of culture. Five scenarios are explored: evolution of independent cultural elements, stepwise modification of elements, differentiation or combination of elements and systems of cultural elements. As one application of our framework, we study the evolution of cultural diversity (in time as well as between groups).

  17. A Model of Culture in Trading Agents

    NARCIS (Netherlands)

    Hofstede, G.J.; Jonker, C.M.; Verwaart, T.

    2013-01-01

    Geert Hofstede’s five-dimensional framework is widely used in social sciences and management science to characterize cultures. It has been suggested to build culturally consistent agent characters based on his framework. This chapter stresses the relevance of culture and trust for trade, substantiat

  18. Spiral vane bioreactor

    Science.gov (United States)

    Morrison, Dennis R. (Inventor)

    1991-01-01

    A spiral vane bioreactor of a perfusion type is described in which a vertical chamber, intended for use in a microgravity condition, has a central rotating filter assembly and has flexible membranes disposed to rotate annularly about the filter assembly. The flexible members have end portions disposed angularly with respect to one another. A fluid replenishment medium is input from a closed loop liquid system to a completely liquid filled chamber containing microcarrier beads, cells and a fluid medium. Output of spent medium is to the closed loop. In the closed loop, the output and input parameters are sensed by sensors. A manifold permits recharging of the nutrients and pH adjustment. Oxygen is supplied and carbon dioxide and bubbles are removed and the system is monitored and controlled by a microprocessor.

  19. [Transgenic animals bioreactors].

    Science.gov (United States)

    Gou, Ke-Mian; An, Xiao-Rong; Tian, Jian-Hui; Chen, Yong-Fu

    2002-01-01

    The production of human recombinant proteins in milk of transgenic farm animals offers a safe, very cost-effective source of commercially important proteins that cannot be produced as efficiently in adequate quantities by other methods. This review has summarized the current status of gene selection, vector construct, transgenic methods, economics, and obvious potential in transgenic animals bioreactors. Recently, a more powerful approach was adopted in the transgenic animals founded on the application of nuclear transfer. As we will illustrate, this strategy presents a breakthrough in the overall efficiency of generating transgenic farm animals, product consistency, and time of product development. The successful adaptation of Cre-/lox P-mediated site-specific DNA recombination systems in farm animals will offer unprecedented possibilities for generating transgenic animals.

  20. Fast multipoint immobilized MOF bioreactor.

    Science.gov (United States)

    Liu, Wan-Ling; Wu, Cheng-You; Chen, Chien-Yu; Singco, Brenda; Lin, Chia-Her; Huang, Hsi-Ya

    2014-07-14

    An enzyme-NBD@MOF bioreactor with exemplary proteolytic performance, even after successive reuse and storage, was produced through a novel, rapid and simple multipoint immobilization technique without chemical modification of the solid support. Enzyme loading and distribution could be directly monitored from the fluorescence emission of the bioreactor. The dye molecular dimension plays a role in its overall performance. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Review of recent developments in modeling of packed-bed solid-state fermentation bioreactors%填充床固态发酵生物反应器模拟研究进展

    Institute of Scientific and Technical Information of China (English)

    董吉林; 杜冰; 申瑞玲

    2012-01-01

    Mathematical models are important tools for optimizing the design and operation of solid-state fermentation (SSF) bioreactors. Not only can models guide the design and operation of bioreactors but they can also provide insights into how the various phenomena within the fermentation system combine to control overall process performance. The current article evaluates the proposed models of Packed-Bed Solid-State Fermentation Bioreactors that hypothesis the substrate bed to be treated as a single pseudohomogeneous phase or non-pseudohomogeneous phase,and intermittently-mixed packed-bed bioreactors. The insights that have been achieved through the modeling work and the advances that have been made in heat and mass transportation models are discussed. In addition, the improvements of porous media volume averaging theory to heat and mass transportation models that will be necessary in the future are discussed.%数学模型是固态发酵生物反应嚣设计、优化和操作的重要工具,它不仅可以指导SSF过程生物反应器的设计和操作,而且可以深入分析发酵系统内的各种现象从而控制整个发酵过程.文章分析近几年固态发酵填充床生物反应器的拟均相假设模拟模型、非均相假设模拟模型和间歇混合填充床生物反应器模拟模型,对模拟结果进行讨论,深入分析如何用数学方法描述SSF系统的传热传质模型.在分析填充床固态发酵底物床层的系统组成及其性质的基础上,提出利用多孔介质体积平均理论研究SSF系统传热传质过程的可能性.

  2. Modeling of 1,2-Dichloroethane Biodegradation by Xanthobacter autotrophicus GJ10 under Shock Loading of Other Halogenated Compounds in Continuous Stirred Tank Bioreactor

    NARCIS (Netherlands)

    Beschkov, V.; Sapundzhiev, Ts.; Torz, M.; Wietzes, P.; Janssen, D. B.

    2008-01-01

    A mathematical model describing the behavior of a continuous culture that degrades 1,2-dichloroethane and receives a shock loading of another compound was developed. The model takes into account possible cell death due to toxicity, growth inhibition and additional growth of cells on the second carbo

  3. 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

  4. Verification of energy dissipation rate scalability in pilot and production scale bioreactors using computational fluid dynamics.

    Science.gov (United States)

    Johnson, Chris; Natarajan, Venkatesh; Antoniou, Chris

    2014-01-01

    Suspension mammalian cell cultures in aerated stirred tank bioreactors are widely used in the production of monoclonal antibodies. Given that production scale cell culture operations are typically performed in very large bioreactors (≥ 10,000 L), bioreactor scale-down and scale-up become crucial in the development of robust cell-culture processes. For successful scale-up and scale-down of cell culture operations, it is important to understand the scale-dependence of the distribution of the energy dissipation rates in a bioreactor. Computational fluid dynamics (CFD) simulations can provide an additional layer of depth to bioreactor scalability analysis. In this communication, we use CFD analyses of five bioreactor configurations to evaluate energy dissipation rates and Kolmogorov length scale distributions at various scales. The results show that hydrodynamic scalability is achievable as long as major design features (# of baffles, impellers) remain consistent across the scales. Finally, in all configurations, the mean Kolmogorov length scale is substantially higher than the average cell size, indicating that catastrophic cell damage due to mechanical agitation is highly unlikely at all scales.

  5. Development of a hybrid scaffold and a bioreactor for cartilage regeneration

    Institute of Scientific and Technical Information of China (English)

    LEE Seung-Jae; LEE In Hwan; PARK Jeong Hun; GWAK So-Jung; RHIE Jong-Won; CHO Dong-Woo; KO Tae Jo; KIM Dong Sung

    2009-01-01

    We developed a hybrid scaffold and a bioreactor for cartilage regeneration. The hybrid scaffold was developed as combination of two components: a biodegradable framework and hydrogel-containing chondrocytes. We performed the MTT cell proliferation assay to compare the proliferation and viability of chondrocytes on three types of scaffolds: an alginate gel, the hybrid scaffold, and an alginate sponge. Cells were encapsulated in 2% agarose gel. The bioreactor consisted of a circulation system and a compression system. We performed dynamic cell culture on these agarose gels in the bioreactor for 3 days.

  6. University - industry collaborations: models, drivers and cultures.

    Science.gov (United States)

    Ehrismann, Dominic; Patel, Dhavalkumar

    2015-01-01

    The way academic institutions and pharmaceutical companies have been approaching collaborations has changed significantly in recent years. A multitude of interaction models were tested and critical factors that drive successful collaborations have been proposed. Based on this experience the current consensus in the pharmaceutical industry is to pursue one of two strategies: an open innovation approach to source discoveries wherever they occur, or investing selectively into scientific partnerships that churn out inventions that can be translated from bench to bedside internally. While these strategies may be intuitive, to form and build sustainable relationships between academia and large multinational healthcare enterprises is proving challenging. In this article we explore some of the more testing aspects of these collaborations, approaches that various industrial players have taken and provide our own views on the matter. We found that understanding and respecting each other's organisational culture and combining the intellectual and technological assets to answer big scientific questions accelerates and improves the quality of every collaboration. Upon discussing the prevailing cooperation models in the university - industry domain, we assert that science-driven collaborations where risks and rewards are shared equally without a commercial agenda in mind are the most impactful.

  7. Modeling of filtrate COD in submerged membrane bioreactor for oily wastewater treatment%SMBR处理含油废水出水COD数学模型

    Institute of Scientific and Technical Information of China (English)

    杜晓文; 王占生; 满春志; 李薇; 刘磊; 徐毅

    2012-01-01

    根据微生物生长动力学特征以及膜分离特征,建立恒通量下运行的一体式膜生物反应器系统出水COD数学模型,提出膜生物反应器处理效率的数学模型。以实验及模型为基础,分别对进水COD浓度控制在300、400、500 mg/L附近时经过反应器后COD的去除效率进行了比较。通过公式计算的数据和实验数据分析可得:COD去除率的公式计算值与实验结果比较吻合,相对偏差仅为0.0223,为膜系统有机物的去除效果估算提供了基础,可为该类工艺的参数选择与优化提供参考。%To predict the effluent quality in MBR processes, a mathematic model for removal efficiency in MBR processes was established, deriving from models standing for the chemical oxygen demand (COD) of the ef- fluent with constant membrane flux. The model was also based on the dynamics characteristics of microbial growth and membrane separation. The experiment was carried out in a lab-scale to treat oily wastewater in a submerged membrane bioreactor (SMBR). Thereby comparisons between experimental data and imputed value were investiga- ted at COD values of 300,400 and 500 mg/L. The results showed data good match of the COD removal efficiency between models and experiment, with a relative deviation of O. 0223, providing a foundation for estimating organics removing efficiency in membrane system and a recommendation for preferences and optimization of the processes.

  8. BIOREACTOR ECONOMICS, SIZE AND TIME OF OPERATION (BEST) COMPUTER SIMULATOR FOR DESIGNING SULFATE-REDUCING BACTERIA FIELD BIOREACTORS

    Science.gov (United States)

    BEST (bioreactor economics, size and time of operation) is an Excel™ spreadsheet-based model that is used in conjunction with the public domain geochemical modeling software, PHREEQCI. The BEST model is used in the design process of sulfate-reducing bacteria (SRB) field bioreacto...

  9. The Culture-Work-Health Model and Work Stress.

    Science.gov (United States)

    Peterson, Michael; Wilson, John F.

    2002-01-01

    Examines the role of organizational culture in the etiology of workplace stress through the framework of the Culture-Work- Health model. A review of relevant business and health literature indicates that culture is an important component of work stress and may be a key to creating effective organizational stress interventions. (SM)

  10. Social Importance Dynamics: A Model for Culturally-Adaptive Agents

    NARCIS (Netherlands)

    Mascarenhas, S.; Prada, R.; Paiva, A.; Hofstede, G.J.

    2013-01-01

    The unwritten rules of human cultures greatly affect social behaviour and as such should be considered in the development of socially intelligent agents. So far, there has been a large focus on modeling cultural aspects related to non-verbal behaviour such as gaze or body posture. However, culture a

  11. The Culture-Work-Health Model and Work Stress.

    Science.gov (United States)

    Peterson, Michael; Wilson, John F.

    2002-01-01

    Examines the role of organizational culture in the etiology of workplace stress through the framework of the Culture-Work- Health model. A review of relevant business and health literature indicates that culture is an important component of work stress and may be a key to creating effective organizational stress interventions. (SM)

  12. Hydrodynamic study to the bioreactor at the Granollers wastewater plant; Estudio hidrodinamico del reactor biologia de la EDAR Granollers

    Energy Technology Data Exchange (ETDEWEB)

    Olivet Santana, D.; Valls Puig, J.; Gordillo Bolasell, M. A.; Sanchez Ferrer, A.; Freixo Rey, A.

    2002-07-01

    One of the most important parameters affecting the operation of bioreactors used in the wastewater treatment is their hydrodynamic behaviour. Usually, bioreactors are designed according to ideal flow models, concretely stirred tank and plug flow models. However, actual bioreactors show a different behaviour from that predicted from ideal modelling. This fact is due to the presence of multiphase systems (e. g. biomass, liquid phase, air, etc.) and the yield obtained can significantly differ from theoretical design values. In the present work, residence-time distribution (RTD) is used to study a plug-flow bioreactor, which is currently working at the plant placed in Granollers (Barcelona). (Author) 5 refs.

  13. 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.

  14. Residential segregation and cultural dissemination: An Axelrod-Schelling model

    Science.gov (United States)

    Gracia-Lázaro, C.; Lafuerza, L. F.; Floría, L. M.; Moreno, Y.

    2009-10-01

    In the Axelrod’s model of cultural dissemination, we consider the mobility of cultural agents through the introduction of a density of empty sites and the possibility that agents in a dissimilar neighborhood can move to them if their mean cultural similarity with the neighborhood is below some threshold. While for low values of the density of empty sites, the mobility enhances the convergence to a global culture, for high enough values of it, the dynamics can lead to the coexistence of disconnected domains of different cultures. In this regime, the increase in initial cultural diversity paradoxically increases the convergence to a dominant culture. Further increase in diversity leads to the fragmentation of the dominant culture into domains, forever changing in shape and number, as an effect of the never ending eroding activity of cultural minorities.

  15. Cultural Resources as Sustainability Enablers: Towards a Community-Based Cultural Heritage Resources Management (COBACHREM Model

    Directory of Open Access Journals (Sweden)

    Susan O. Keitumetse

    2013-12-01

    Full Text Available People inhabit and change environments using socio-cultural and psycho-social behaviors and processes. People use their socio-cultural understanding of phenomena to interact with the environment. People are carriers of cultural heritage. These characteristics make cultural values ubiquitous in all people-accessed and people-inhabited geographic spaces of the world, making people readily available assets through which environmental sustainability can be implemented. Yet, people’s conservation development is rarely planned using cultural resources. It is against this background that a Community-Based Cultural Heritage Resources Management (COBACHREM model is initiated as a new approach that outlines the symbiosis between cultural heritage, environment and various stakeholders, with a view to create awareness about neglected conservation indicators inherent in cultural resources and better placed to complement already existing natural resources conservation indicators. The model constitutes a two-phased process with four (04 levels of operation, namely: level I (production; level II (reproduction; level III (consumption that distinguish specific components of cultural heritage resources to be monitored at level IV for sustainability using identified cultural conservation indicators. Monitored indicators, which are limitless, constitute work in progress of the model and will be constantly reviewed, renewed and updated through time. Examples of monitoring provided in this article are the development of cultural competency-based training curriculum that will assist communities to transform cultural information into certifiable intellectual (educational and culture-economic (tourism assets. Another monitoring example is the mainstreaming of community cultural qualities into already existing environmental conservation frameworks such as eco-certification to infuse new layers of conservation indicators that enrich resource sustainability. The technical

  16. Cultural Policy Implementation: Theoretical Conceptualization and Models

    National Research Council Canada - National Science Library

    Audrone Pauliukeviciute; Alvydas Raipa

    2014-01-01

      Rapidly changing global and technological world, continuous progressive change and cultural transformations determine changes in country's management systems, in separate spheres of public policy and management...

  17. Enhancing inhibited fermentations through a dynamic electro-membrane bioreactor

    DEFF Research Database (Denmark)

    Prado Rubio, Oscar Andres; Garde, Arvid; Rype, Jens-Ulrik

    its strong potential for increasing productivity and product yield has been verified. REED uses ion exchange membranes and electrical potential gradients to selectively separate the target ion. The main limitation of using membrane separation combined with bioreactors is membrane fouling. REED...... technology ensures long operation time by reversing periodically the polarity of the imposed electrical field to significantly reduce the influence of membrane fouling. The periodic nature of the electrically driven membrane separation process makes the membrane bioreactor operation non trivial....... This challenging operation is associated with different dynamic behaviors of the individual units plus their interaction. The purpose of this contribution is to show the results of experimental and model based efforts done in order to investigate the operation of a membrane bioreactor. From modeling point of view...

  18. Development of a Laminar Flow Bioreactor by Computational Fluid Dynamics

    Directory of Open Access Journals (Sweden)

    Meir Israelowitz

    2012-01-01

    Full Text Available The purpose of this study is to improve the design of a bioreactor for growing bone and other three-dimensional tissues using a computational fluid dynamics (CFD software to simulate flow through a porous scaffold, and to recommend design changes based on the results. Basic requirements for CFD modeling were that the flow in the reactor should be laminar and any flow stagnation should be avoided in order to support cellular growth within the scaffold. We simulated three different designs with different permeability values of the scaffold and tissue. Model simulation addressed flow patterns in combination with pressure distribution within the bioreactor. Pressure build-up and turbulent flow within the reactor was solved by introduction of an integrated bypass system for pressure release. The use of CFD afforded direct feedback to optimize the bioreactor design.

  19. CULTURE ET MANAGEMENT: LE MODEL D’HOFSTEDE EN QUESTION

    Directory of Open Access Journals (Sweden)

    Soufyane BADRAOUI

    2014-12-01

    Full Text Available Extensive research has been conducted looking at the links between culture and management. Studies have examined, among other components of the managerial activity captured by power distance, individualism, etc. However; these studies assume the existence of a national culture, something largely absent in most countries that are dominated by the cultures of their various ethnic communities. Different cultures perceive the benefits differently,found significant differences between cultures in perceptions and organizational/ Managerial practices. Our modest contribution is structured in two parts: the first part concerns the presentation of the Hofstede model and analysis through this model on some African societies, the second reserved for discussing expose in socio-cultural characters that influences the African Management and limitations of the model of cultural dimensions of Hofstede and conclusion.

  20. A Culture-Behavior-Brain Loop Model of Human Development.

    Science.gov (United States)

    Han, Shihui; Ma, Yina

    2015-11-01

    Increasing evidence suggests that cultural influences on brain activity are associated with multiple cognitive and affective processes. These findings prompt an integrative framework to account for dynamic interactions between culture, behavior, and the brain. We put forward a culture-behavior-brain (CBB) loop model of human development that proposes that culture shapes the brain by contextualizing behavior, and the brain fits and modifies culture via behavioral influences. Genes provide a fundamental basis for, and interact with, the CBB loop at both individual and population levels. The CBB loop model advances our understanding of the dynamic relationships between culture, behavior, and the brain, which are crucial for human phylogeny and ontogeny. Future brain changes due to cultural influences are discussed based on the CBB loop model.

  1. 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 wastewater

  2. Modelling Trade and Trust Across Cultures

    NARCIS (Netherlands)

    Hofstede, G.J.; Jonker, C.M.; Meijer, S.; Verwaart, D.

    2006-01-01

    Misunderstandings arise in international trade due to difference in cultural background of trade partners. Trust and the role it plays in trade are influenced by culture. Considering that trade always involves working on the relationship with the trade partner, understanding the behaviour of the oth

  3. Modeling cultural behavior for military virtual training

    NARCIS (Netherlands)

    Kerbusch, P.; Schram, J.; Bosch, K. van den

    2011-01-01

    Soldiers on mission in areas with unfamiliar cultures must be able to take into account the norms of the local culture when assessing a situation, and must be able to adapt their behavior accordingly. Innovative technologies provide opportunity to train the required skills in an interactive and real

  4. Modeling Cultural Behavior for Military Virtual Training

    NARCIS (Netherlands)

    Bosch, K. van den; Kerbusch, P.J.M.; Schram, J.

    2012-01-01

    Soldiers on mission in areas with unfamiliar cultures must be able to take into account the norms of the local culture when assessing a situation, and must be able to adapt their behavior accordingly. Innovative technologies provide opportunity to train the required skills in an interactive and real

  5. Parents as Models in Children's Cultural Development.

    Science.gov (United States)

    Swick, Kevin J.

    1986-01-01

    Defines cross-cultural parent performance roles and discusses common goals. Outlines criteria among diverse parenting styles and lists areas of parental functioning for self-image determination status. Provides guidelines for assessing parent cultural competence and names three major tasks for educators in regard to educating parents for…

  6. Models of cultural niche construction with selection and assortative mating.

    Science.gov (United States)

    Creanza, Nicole; Fogarty, Laurel; Feldman, Marcus W

    2012-01-01

    Niche construction is a process through which organisms modify their environment and, as a result, alter the selection pressures on themselves and other species. In cultural niche construction, one or more cultural traits can influence the evolution of other cultural or biological traits by affecting the social environment in which the latter traits may evolve. Cultural niche construction may include either gene-culture or culture-culture interactions. Here we develop a model of this process and suggest some applications of this model. We examine the interactions between cultural transmission, selection, and assorting, paying particular attention to the complexities that arise when selection and assorting are both present, in which case stable polymorphisms of all cultural phenotypes are possible. We compare our model to a recent model for the joint evolution of religion and fertility and discuss other potential applications of cultural niche construction theory, including the evolution and maintenance of large-scale human conflict and the relationship between sex ratio bias and marriage customs. The evolutionary framework we introduce begins to address complexities that arise in the quantitative analysis of multiple interacting cultural traits.

  7. Models of cultural niche construction with selection and assortative mating.

    Directory of Open Access Journals (Sweden)

    Nicole Creanza

    Full Text Available Niche construction is a process through which organisms modify their environment and, as a result, alter the selection pressures on themselves and other species. In cultural niche construction, one or more cultural traits can influence the evolution of other cultural or biological traits by affecting the social environment in which the latter traits may evolve. Cultural niche construction may include either gene-culture or culture-culture interactions. Here we develop a model of this process and suggest some applications of this model. We examine the interactions between cultural transmission, selection, and assorting, paying particular attention to the complexities that arise when selection and assorting are both present, in which case stable polymorphisms of all cultural phenotypes are possible. We compare our model to a recent model for the joint evolution of religion and fertility and discuss other potential applications of cultural niche construction theory, including the evolution and maintenance of large-scale human conflict and the relationship between sex ratio bias and marriage customs. The evolutionary framework we introduce begins to address complexities that arise in the quantitative analysis of multiple interacting cultural traits.

  8. Statistical Inference of Biometrical Genetic Model With Cultural Transmission.

    Science.gov (United States)

    Guo, Xiaobo; Ji, Tian; Wang, Xueqin; Zhang, Heping; Zhong, Shouqiang

    2013-01-01

    Twin and family studies establish the foundation for studying the genetic, environmental and cultural transmission effects for phenotypes. In this work, we make use of the well established statistical methods and theory for mixed models to assess cultural transmission in twin and family studies. Specifically, we address two critical yet poorly understood issues: the model identifiability in assessing cultural transmission for twin and family data and the biases in the estimates when sub-models are used. We apply our models and theory to two real data sets. A simulation is conducted to verify the bias in the estimates of genetic effects when the working model is a sub-model.

  9. Validation of a model for process development and scale-up of packed-bed solid-state bioreactors

    NARCIS (Netherlands)

    Weber, F.J.; Oostra, J.; Tramper, J.; Rinzema, A.

    2002-01-01

    We have validated our previously described model for scale-up of packed-bed solid-state fermenters (Weber et al., 1999) with experiments in an adiabatic 15-dm3 packed-bed reactor, using the fungi Coniothyrium minitans and Aspergillus oryzae. Effects of temperature on respiration, growth, and sporula

  10. Cross-scale predictive modeling of CHO cell culture growth and metabolites using Raman spectroscopy and multivariate analysis.

    Science.gov (United States)

    Berry, Brandon; Moretto, Justin; Matthews, Thomas; Smelko, John; Wiltberger, Kelly

    2015-01-01

    Multi-component, multi-scale Raman spectroscopy modeling results from a monoclonal antibody producing CHO cell culture process including data from two development scales (3 L, 200 L) and a clinical manufacturing scale environment (2,000 L) are presented. Multivariate analysis principles are a critical component to partial least squares (PLS) modeling but can quickly turn into an overly iterative process, thus a simplified protocol is proposed for addressing necessary steps including spectral preprocessing, spectral region selection, and outlier removal to create models exclusively from cell culture process data without the inclusion of spectral data from chemically defined nutrient solutions or targeted component spiking studies. An array of single-scale and combination-scale modeling iterations were generated to evaluate technology capabilities and model scalability. Analysis of prediction errors across models suggests that glucose, lactate, and osmolality are well modeled. Model strength was confirmed via predictive validation and by examining performance similarity across single-scale and combination-scale models. Additionally, accurate predictive models were attained in most cases for viable cell density and total cell density; however, these components exhibited some scale-dependencies that hindered model quality in cross-scale predictions where only development data was used in calibration. Glutamate and ammonium models were also able to achieve accurate predictions in most cases. However, there are differences in the absolute concentration ranges of these components across the datasets of individual bioreactor scales. Thus, glutamate and ammonium PLS models were forced to extrapolate in cases where models were derived from small scale data only but used in cross-scale applications predicting against manufacturing scale batches. © 2014 American Institute of Chemical Engineers.

  11. Reconstructing marginality: a new model of cultural diversity in nursing.

    Science.gov (United States)

    Southwick, Margaret; Polaschek, Nick

    2014-05-01

    This article presents a new model of cultural diversity in nursing that critically reconstructs the concept of marginality that underpins other models. Rather than viewing the marginal as "other," marginality is redefined as the space in between the dominant cultural reality and the cultural realities of minority groups located within a society. Members of a minority cultural group who become skilled in the difficult process of negotiating this in-between space open the possibility of transformation within nursing education and practice. This model has been applied in a study of the experience of nursing students of Pacific ethnicity in New Zealand. Subsequently, an undergraduate Pacific nursing program was developed, with greatly increased success rates in registration of Pacific nurses. This model of cultural diversity can also be used to understand nursing practice involving people from minority cultures or other socially excluded categories.

  12. Three-dimensional cell culture models for investigating human viruses.

    Science.gov (United States)

    He, Bing; Chen, Guomin; Zeng, Yi

    2016-10-01

    Three-dimensional (3D) culture models are physiologically relevant, as they provide reproducible results, experimental flexibility and can be adapted for high-throughput experiments. Moreover, these models bridge the gap between traditional two-dimensional (2D) monolayer cultures and animal models. 3D culture systems have significantly advanced basic cell science and tissue engineering, especially in the fields of cell biology and physiology, stem cell research, regenerative medicine, cancer research, drug discovery, and gene and protein expression studies. In addition, 3D models can provide unique insight into bacteriology, virology, parasitology and host-pathogen interactions. This review summarizes and analyzes recent progress in human virological research with 3D cell culture models. We discuss viral growth, replication, proliferation, infection, virus-host interactions and antiviral drugs in 3D culture models.

  13. 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.

    Science.gov (United States)

    Leferink, Anne M; Chng, Yhee-Cheng; van Blitterswijk, Clemens A; Moroni, Lorenzo

    2015-01-01

    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 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.

  14. 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.

  15. Biological removal of nitrate by an oil reservoir culture capable of autotrophic and heterotrophic activities: kinetic evaluation and modeling of heterotrophic process.

    Science.gov (United States)

    An, Shijie; Stone, Heather; Nemati, Mehdi

    2011-06-15

    Kinetics of heterotrophic denitrification was investigated using an oil reservoir culture with the ability to function under both autotrophic and heterotrophic conditions. In the batch system nitrate at concentrations up to 30 mM did not influence the kinetics but with 50mM slower growth and removal rates were observed. A kinetic model, representing the denitrification as reduction of nitrate to nitrite, and subsequent reduction of nitrite to nitrous oxides and nitrogen gas was developed. The value of various kinetic coefficients, including maximum specific growth rate, saturation constant, yield and activation energy for nitrate and nitrite reductions were determined by fitting the experimental data into the developed model. In continuous bioreactors operated with 10 or 30 mM nitrate, complete removal of nitrate (no residual nitrite) and linear dependency between nitrate loading and removal rates were observed for loading rates up to 0.21 and 0.58 mM h(-1), respectively. The highest removal rates of 0.31 and 0.94 mM h(-1) observed at loading rates of 0.42 mM h(-1) and 1.26 mM h(-1), with corresponding removal percentages of nitrate and total nitrogen being 75.4, 54.4%, and 74.4 and 17.9%, respectively. Developed kinetic model predicted the performance of the continuous bioreactors with accuracy.

  16. Membrane bioreactor for waste gas treatment

    NARCIS (Netherlands)

    Reij, M.W.

    1997-01-01

    Summary

    This thesis describes the design and testing of a membrane bioreactor (MBR) for removal of organic pollutants from air. In such a bioreactor for biological gas treatment pollutants are degraded by micro-organisms. The membrane bioreactor is an alternative to other types of

  17. Preliminary Study on Airlift Membran—Bioreactor

    Institute of Scientific and Technical Information of China (English)

    XUNong; XINGWeihong; 等

    2002-01-01

    A new type of membrane bioreactor named “airlift membrane-bioreactor”is discussed.For municipal wastewater reclamation,the preliminary study on airlift membrane-bioreactor shows its good performance such as high flux and lower energy consumption.The airlift membrane-bioreactor is potentially applicable in bioengineering and environmental protection fields.

  18. Two-step nitrification in a pure moving bed biofilm reactor-membrane bioreactor for wastewater treatment: nitrifying and denitrifying microbial populations and kinetic modeling.

    Science.gov (United States)

    Leyva-Díaz, J C; González-Martínez, A; Muñío, M M; Poyatos, J M

    2015-12-01

    The moving bed biofilm reactor-membrane bioreactor (MBBR-MBR) is a novel solution to conventional activated sludge processes and membrane bioreactors. In this study, a pure MBBR-MBR was studied. The pure MBBR-MBR mainly had attached biomass. The bioreactor operated with a hydraulic retention time (HRT) of 9.5 h. The kinetic parameters for heterotrophic and autotrophic biomasses, mainly nitrite-oxidizing bacteria (NOB), were evaluated. The analysis of the bacterial community structure of the ammonium-oxidizing bacteria (AOB), NOB, and denitrifying bacteria (DeNB) from the pure MBBR-MBR was carried out by means of pyrosequencing to detect and quantify the contribution of the nitrifying and denitrifying bacteria in the total bacterial community. The relative abundance of AOB, NOB, and DeNB were 5, 1, and 3%, respectively, in the mixed liquor suspended solids (MLSS), and these percentages were 18, 5, and 2%, respectively, in the biofilm density (BD) attached to carriers. The pure MBBR-MBR had a high efficiency of total nitrogen (TN) removal of 71.81±16.04%, which could reside in the different bacterial assemblages in the fixed biofilm on the carriers. In this regard, the kinetic parameters for autotrophic biomass had values of YA=2.3465 mg O2 mg N(-1), μm, A=0.7169 h(-1), and KNH=2.0748 mg NL(-1).

  19. Propagation and Dissolution of CO2 bubbles in Algae Photo-bioreactors

    Science.gov (United States)

    Kosaraju, Srinivas

    2015-11-01

    Research grade photo-bioreactors are used to study and cultivate different algal species for biofuel production. In an attempt to study the growth properties of a local algal species in rain water, a custom made bioreactor is designed and being tested. Bio-algae consumes dissolved CO2 in water and during its growth cycle, the consumed CO2 must be replenished. Conventional methods use supply of air or CO2 bubbles in the growth medium. The propagation and dissolution of the bubbles, however, are strongly dependent on the design parameters of the photo-bioreactor. In this paper, we discuss the numerical modeling of the air and CO2 bubble propagation and dissolution in the photo-bioreactor. Using the results the bioreactor design will be modified for maximum productivity.

  20. Dynamic Single-Use Bioreactors Used in Modern Liter- and m(3)- Scale Biotechnological Processes: Engineering Characteristics and Scaling Up.

    Science.gov (United States)

    Löffelholz, Christian; Kaiser, Stephan C; Kraume, Matthias; Eibl, Regine; Eibl, Dieter

    2014-01-01

    During the past 10 years, single-use bioreactors have been well accepted in modern biopharmaceutical production processes targeting high-value products. Up to now, such processes have mainly been small- or medium-scale mammalian cell culture-based seed inoculum, vaccine or antibody productions. However, recently first attempts have been made to modify existing single-use bioreactors for the cultivation of plant cells and tissue cultures, and microorganisms. This has even led to the development of new single-use bioreactor types. Moreover, due to safety issues it has become clear that single-use bioreactors are the "must have" for expanding human stem cells delivering cell therapeutics, the biopharmaceuticals of the next generation. So it comes as no surprise that numerous different dynamic single-use bioreactor types, which are suitable for a wide range of applications, already dominate the market today. Bioreactor working principles, main applications, and bioengineering data are presented in this review, based on a current overview of greater than milliliter-scale, commercially available, dynamic single-use bioreactors. The focus is on stirred versions, which are omnipresent in R&D and manufacturing, and in particular Sartorius Stedim's BIOSTAT family. Finally, we examine development trends for single-use bioreactors, after discussing proven approaches for fast scaling-up processes.

  1. Modelling and dynamic simulation of a pilot-scale moving bed bioreactor for the treatment of municipal wastewater: model concepts and the use of respirometry for the estimation of kinetic parameters.

    Science.gov (United States)

    Plattes, M; Fiorelli, D; Gillé, S; Girard, C; Henry, E; Minette, F; O'Nagy, O; Schosseler, P M

    2007-01-01

    A model for the simulation of a moving bed bioreactor (MBBR) used for the treatment of municipal wastewater is proposed. The model includes attachment of particulates to the biofilm and detachment of biofilm into the bulk liquid. The growth kinetics are modelled with the activated sludge model no. 1 (ASM1). Respirometry was used for the estimation of kinetic parameters. The resulting respirograms featured the typical endogenous and exogenous respiration phases and the respirogram shapes were as expected from analogous respirometry with activated sludge. The estimated parameter set was used for modelling and simulation of the pilot-scale MBBR. The main proportion of biomass in the MBBR was found to be attached as biofilm on the carrier elements (4.1 -4.6 g dm-3) and only a small amount was suspended in the bulk liquid (0.15gdm(-3)). Attachment and detachment rates were estimated to be 4.8-7.5g m(-2) d(-1) 1for attachment and 6.5-7.5g m(-2) d(-1) for detachment. The biofilm age was estimated to be 1.8-2.7d. The model was used to predict effluent quality parameters and a good fit of the simulated data to the measured data originating from a four-days-long measurement campaign was obtained.

  2. Cell culture process development: advances in process engineering.

    Science.gov (United States)

    Heath, Carole; Kiss, Robert

    2007-01-01

    Representatives from the cell culture process development community met on September 11 and 12, 2006 at the ACS National Meeting in San Francisco to discuss "Cell Culture Process Development: Advances in Process Engineering". This oral session was held as part of the Division of Biochemical Technology (BIOT) program. The presentations addressed the very small scale (less than 1 mL) to the very large scale (20,000 L). The topics covered included development of high throughput cell culture screening systems, modeling and characterization of bioreactor environments from mixing and shear perspectives at both small and large scales, systematic approaches for improving scale-up and scale-down activities, development of disposable bioreactor technologies, and novel perfusion culture approaches. All told, this well-attended session resulted in a valuable exchange of technical information and demonstrated a high level of interest within the process development community.

  3. A calibration protocol of a one-dimensional moving bed bioreactor (MBBR) dynamic model for nitrogen removal.

    Science.gov (United States)

    Barry, U; Choubert, J-M; Canler, J-P; Héduit, A; Robin, L; Lessard, P

    2012-01-01

    This work suggests a procedure to correctly calibrate the parameters of a one-dimensional MBBR dynamic model in nitrification treatment. The study deals with the MBBR configuration with two reactors in series, one for carbon treatment and the other for nitrogen treatment. Because of the influence of the first reactor on the second one, the approach needs a specific calibration strategy. Firstly, a comparison between measured values and simulated ones obtained with default parameters has been carried out. Simulated values of filtered COD, NH(4)-N and dissolved oxygen are underestimated and nitrates are overestimated compared with observed data. Thus, nitrifying rate and oxygen transfer into the biofilm are overvalued. Secondly, a sensitivity analysis was carried out for parameters and for COD fractionation. It revealed three classes of sensitive parameters: physical, diffusional and kinetic. Then a calibration protocol of the MBBR dynamic model was proposed. It was successfully tested on data recorded at a pilot-scale plant and a calibrated set of values was obtained for four parameters: the maximum biofilm thickness, the detachment rate, the maximum autotrophic growth rate and the oxygen transfer rate.

  4. The Fluid Mechanics of a Wavy-Wall Bioreactor

    Science.gov (United States)

    Sucosky, Philippe; Bilgen, Bahar; Aleem, Alexander; Neitzel, Paul; Barabino, Gilda

    2004-11-01

    Bioreactors are devices used for the production of mammalian tissue in vitro. Although mixing has been shown to stimulate the growth of cartilage constructs, high shear-stress levels can damage the cells. In order to enhance mixing while minimizing shear, a wavy-wall bioreactor (WWB) featuring a sinusoidal internal profile has been designed. The turbulent hydrodynamic environment produced in this device is investigated experimentally using particle-image velocimetry. A model bioreactor made of acrylic and filled with an index-matching solution of zinc iodide is used to compensate for the refraction of light at the walls. The flow observed in different planes is shown to be periodic, spatially dependent, and dominated by mean-shear rather than Reynolds stresses in the vicinity of constructs. Finally, a comparison between the mean-shear stresses obtained in the WWB and in a standard spinner flask reveals similar stress levels near the construct walls.

  5. A Culture-Sensitive Agent in Kirman's Ant Model

    Science.gov (United States)

    Chen, Shu-Heng; Liou, Wen-Ching; Chen, Ting-Yu

    The global financial crisis brought a serious collapse involving a "systemic" meltdown. Internet technology and globalization have increased the chances for interaction between countries and people. The global economy has become more complex than ever before. Mark Buchanan [12] indicated that agent-based computer models will prevent another financial crisis and has been particularly influential in contributing insights. There are two reasons why culture-sensitive agent on the financial market has become so important. Therefore, the aim of this article is to establish a culture-sensitive agent and forecast the process of change regarding herding behavior in the financial market. We based our study on the Kirman's Ant Model[4,5] and Hofstede's Natational Culture[11] to establish our culture-sensitive agent based model. Kirman's Ant Model is quite famous and describes financial market herding behavior from the expectations of the future of financial investors. Hofstede's cultural consequence used the staff of IBM in 72 different countries to understand the cultural difference. As a result, this paper focuses on one of the five dimensions of culture from Hofstede: individualism versus collectivism and creates a culture-sensitive agent and predicts the process of change regarding herding behavior in the financial market. To conclude, this study will be of importance in explaining the herding behavior with cultural factors, as well as in providing researchers with a clearer understanding of how herding beliefs of people about different cultures relate to their finance market strategies.

  6. Sensor equipment for quantification of spatial heterogeneity in large bioreactor

    DEFF Research Database (Denmark)

    Nørregaard, Anders; Formenti, Luca Riccardo; Stocks, Stuart M.

    of sensors and in order to apply more sensor equipment the bioreactor has to be modified which is both costly and results in production downtime. The presence of three phases (gas, liquid, and solid), and the opaque nature of the fermentation broth together with the necessity of heat sterilization further...... of the bioreactor. The method leaves a minimal footprint and can be applied to running production to gather large scale fermentation data, without the need of dedicated experimental cultivations. Ultimately, data describing the spatial heterogeneity can be used to enhance existing process models and to create...

  7. Bioreactors for removing methyl bromide following contained fumigations

    Science.gov (United States)

    Miller, L.G.; Baesman, S.M.; Oremland, R.S.

    2003-01-01

    Use of methyl bromide (MeBr) as a quarantine, commodity, or structural fumigant is under scrutiny because its release to the atmosphere contributes to the depletion of stratospheric ozone. A closed-system bioreactor consisting of 0.5 L of a growing culture of a previously described bacterium, strain IMB-1, removed MeBr (> 110 ??mol L-1) from recirculating air. Strain IMB-1 grew slowly to high cell densities in the bioreactor using MeBr as its sole carbon and energy source. Bacterial oxidation of MeBr produced CO2 and hydrobromic acid (HBr), which required continuous neutralization with NaOH for the system to operate effectively. Strain IMB-1 was capable of sustained oxidation of large amounts of MeBr (170 mmol in 46 d). In an open-system bioreactor (10-L fermenter), strain IMB-1 oxidized a continuous supply of MeBr (220 ??mol L-1 in air). Growth was continuous, and 0.5 mol of MeBr was removed from the air supply in 14 d. The specific rate of MeBr oxidation was 7 ?? 10-16 mol cell-1 h-1. Bioreactors such as these can therefore be used to remove large quantities of contaminant MeBr, which opens the possibility of biodegradation as a practical means for its disposal.

  8. Validation of a Business Model for Cultural Heritage Institutions

    Directory of Open Access Journals (Sweden)

    Cristian CIUREA

    2015-01-01

    Full Text Available The paper proposes a business model for the efficiency optimization of the interaction between all actors involved in cultural heritage sector, such as galleries, libraries, archives and museums (GLAM. The validation of the business model is subject of analyses and implementations in a real environment made by different cultural institutions. The implementation of virtual exhibitions on mobile devices is described and analyzed as a key factor for increasing the cultural heritage visibility. New perspectives on the development of virtual exhibitions for mobile devices are considered. A study on the number of visitors of cultural institutions is carried out and ways to increase the number of visitors are described.

  9. Fluid Dynamic Modeling to Support the Development of Flow-based Hepatocyte Culture Systems for Metabolism Studies

    Directory of Open Access Journals (Sweden)

    Jenny M Pedersen

    2016-09-01

    Full Text Available Accurate prediction of metabolism is a significant outstanding challenge in toxicology. The best predictions are based on experimental data from in vitro systems using primary hepatocytes. The predictivity of the primary hepatocyte-based culture systems, however, is still limited due to well-known phenotypic instability and rapid decline of metabolic competence within a few hours. Dynamic flow bioreactors for three-dimensional cell cultures are thought to be better at recapitulating tissue microenvironments and show potential to improve in vivo extrapolations of chemical or drug toxicity based on in vitro test results. These more physiologically relevant culture systems hold potential for extending metabolic competence of primary hepatocyte cultures as well. In this investigation, we used computational fluid dynamics (CFD to determine the optimal design of a flow-based hepatocyte culture system for evaluating chemical metabolism in vitro. The main design goals were 1 minimization of shear stress experienced by the cells to maximize viability, 2 rapid establishment of a uniform distribution of test compound in the chamber, and 3 delivery of sufficient oxygen to cells to support aerobic respiration. Two commercially available flow devices — RealBio® (RB and QuasiVivo® (QV — and a custom developed fluidized bed (FB bioreactor were simulated, and turbulence, flow characteristics, test compound distribution, oxygen distribution, and cellular oxygen consumption were analyzed. Experimental results from the bioreactors were used to validate the simulation results.Our results indicate that maintaining adequate oxygen supply is the most important factor to the long-term viability of liver bioreactor cultures. Cell density and system flow patterns were the major determinants of local oxygen concentrations. The experimental results closely corresponded to the in silico predictions. Of the three bioreactors examined in this study, we were able

  10. Fluid Dynamic Modeling to Support the Development of Flow-Based Hepatocyte Culture Systems for Metabolism Studies

    Science.gov (United States)

    Pedersen, Jenny M.; Shim, Yoo-Sik; Hans, Vaibhav; Phillips, Martin B.; Macdonald, Jeffrey M.; Walker, Glenn; Andersen, Melvin E.; Clewell, Harvey J.; Yoon, Miyoung

    2016-01-01

    Accurate prediction of metabolism is a significant outstanding challenge in toxicology. The best predictions are based on experimental data from in vitro systems using primary hepatocytes. The predictivity of the primary hepatocyte-based culture systems, however, is still limited due to well-known phenotypic instability and rapid decline of metabolic competence within a few hours. Dynamic flow bioreactors for three-dimensional cell cultures are thought to be better at recapitulating tissue microenvironments and show potential to improve in vivo extrapolations of chemical or drug toxicity based on in vitro test results. These more physiologically relevant culture systems hold potential for extending metabolic competence of primary hepatocyte cultures as well. In this investigation, we used computational fluid dynamics to determine the optimal design of a flow-based hepatocyte culture system for evaluating chemical metabolism in vitro. The main design goals were (1) minimization of shear stress experienced by the cells to maximize viability, (2) rapid establishment of a uniform distribution of test compound in the chamber, and (3) delivery of sufficient oxygen to cells to support aerobic respiration. Two commercially available flow devices – RealBio® and QuasiVivo® (QV) – and a custom developed fluidized bed bioreactor were simulated, and turbulence, flow characteristics, test compound distribution, oxygen distribution, and cellular oxygen consumption were analyzed. Experimental results from the bioreactors were used to validate the simulation results. Our results indicate that maintaining adequate oxygen supply is the most important factor to the long-term viability of liver bioreactor cultures. Cell density and system flow patterns were the major determinants of local oxygen concentrations. The experimental results closely corresponded to the in silico predictions. Of the three bioreactors examined in this study, we were able to optimize the experimental

  11. The effectiveness of a novel cartridge-based bioreactor design in supporting liver cells.

    Science.gov (United States)

    Niu, Mei; Hammond, Paul; Coger, Robin N

    2009-10-01

    Th