- Email: [email protected]
Stress during cell culture processes
Abstracts / Journal of Biotechnology 136S (2008) S114–S117
IL-010
IL-034
Development of bioartificial liver system and potential of stem cells as hepatocyte source
Stress during cell culture processes
Jung-Keug Park 1,2,∗ , Hee-Hoon Yoon 1 , Bo-Young Jung 1 , Ji-Hyun Lee 3 , Suk-Koo Lee 4 , Young-Jin Kim 5 1
Tissue Engineering and Regenerative Medicine Laboratory, Department of Chemical and Biochemical Engineering, Republic of Korea 2 Wave Energy Bioactivation Technology Center, Department of Medical Biotechnology, Dongguk University, Seoul 100-715, Republic of Korea 3 Tissue Engineering Laboratory, Samsung Biomedical Research Institute, Republic of Korea 4 Department of Surgery, Samsung Medical Center, Seoul 135-710, Republic of Korea 5 Cell Therapy Center, Lifecord Inc., Suwon 442-749, Republic of Korea
S115
Joseph Boudrant ∗ , Naziha Barbouche, Nasir Mehmood, Kenza Boulaya, Marie-Franc¸oise Clincke, Eric Olmos, Stéphane Delaunay, Jean-Louis Goergen, Emmanuel Guédon, Annie Marc Laboratory of Chemical Engineering Sciences, UPR CNRS 6811, NancyUniversité, INPL, 2, avenue de la Forêt de Haye, BP 172, 54505 Vandoeuvre-les-Nancy, France E-mail address: [email protected] (J. Boudrant).
Tissue engineering (TE) is an interdisciplinary field that applies medicine, life science, and engineering to develop biological tissues in vitro (Langer and Vacanti, 1993). In Korea, basic researches on TE have grown rapidly last decade. Now, some products are being launched in market and some are under investigational new drug (IND) application. Apart from other TE products, bioartificial liver (BAL) system is a hybrid of biological drug and medical device. To treat fulminant hepatic failure (FHF) patients, various extracorporeal BAL systems have been developed (van de Kerkhove et al., 2004; Park and Lee, 2005). The performance of the BAL depends on the functional activities of the hepatocytes immobilized in the bioreactor system. A packed-bed bioreactor with gel beads has higher surface area to volume ratio and cell capacity than those of a hollow fiber bioreactor. Therefore, the type of bioreactor may be a better alternative for hollow fiber-based BAL systems. In this study, we developed a stable packed-bed bioreactor system that does not induce any damage to the gel beads in the bioreactor. The bioreactor packed with gel bead containing hepatocyte spheroids perfused with down-ward flow of media in which the flow was generated by difference of a media level between the reservoir and outlet chamber placed before and after the bioreactor, respectively. The reactor showed stable liver functions for 12 h, and any damage was not detected at the beads taken from the bioreactor after the long-term operation. And also to acquire human hepatocyte source in the near future, we have checked which stem cells have the hepatic potential and which culture condition can induce them into functional hepatocytes. Now we think that umbilical cord-derived mesenchymal stem cell (UC-MSC) can be a good candidate as human hepatocyte source according to preliminary results.
During cell cultivation, mainly when in suspension, cells generally encounter or have to support various stresses, either physical or chemical. When in liquid medium physical stresses origin can be for example temperature, tensio-active agents, shear stress, light. For cell storage or solid-state fermentation, physical stress can be lack of water. Chemical stresses can be due to variations of pH, of metabolite concentrations, to added chemicals, oxygen, to formed carbon dioxide. The consequences of these stresses can be analyzed in term of viability, of physiological behaviour, for example regarding the produced metabolite, and of genomic (genome-wide expression analyses). This presentation proposes an overview on this large problematic applied to industrial bioprocesses and then focuses on different industrial bacterial fermentations and one animal cell cultivation. The first is producing the amino-acid glutamic acid using a thermoinducible strain of Corynebacterium glutamicum with the effects on the enveloppe and metabolism. The second concerns one filamentous bacteria (Streptomyces prystinaespiralis) producing antibiotics. The third is producing lactic acid with Lactobacillus casei and the study concerns the influence of agitation on the global productivity. The concerned animal cell will be one CHO strain and the study is about the effect of hydrodynamic and medium composition stresses on its global physiological response. This could be also applied to yeast (Saccharomyces cerevisiae) and microalgae cultivations. Moreover, because of large size bioreactors, heterogeneity problems may occur, particularly for temperature or hydrodynamic stress. So, for a serious optimization of the corresponding bioprocesses, the stress distributions in time and space should be seriously quantified. To do this, the local characterization of stresses is expected to bring a more detailed information (quantification, stress duration) than a global one. This implies the use of sophisticated tools (CFP, PIV or LDV). This illustrates that this way of analyzing bioprocesses and their performances is not an easy task, but it could gain more and more importance in the future.
References
doi:10.1016/j.jbiotec.2008.07.371
E-mail address: [email protected] (J.-K. Park).
Langer, R., Vacanti, J.P., 1993. Tissue engineering. Science 260, 920–926. Park, J.K., Lee, D.H., 2005. Bioartificial liver systems; current status and future perspective. J. Biosci. Bioeng. 99, 311–319. van de Kerkhove, M.P., Hoekstra, R., Chamuleau, R.A., van Gulik, T.M., 2004. Clinical application of bioartificial liver support systems. Ann. Surg. 240, 216–230.
doi:10.1016/j.jbiotec.2008.07.370
IL-036 Design, prototyping and operation of disposable bioreactor systems for stem cell cultivation and tissue formation Uwe Marx ∗ , Christian Demmler, Annika Lubitz, Christoph Giese ProBioGen Ag, Goethestrasse, 54 13086 Berlin, Germany E-mail address: [email protected] (U. Marx). Significant progress has been achieved last 20 years in the area of in vitro tissue culture technologies (Marx, 2007). A number of human organ properties could be successfully emulated in vitro (Griffith and Swartz, 2006). Established human skin equivalents, for example, are featuring the evaluation of dermal transfer,
IL-010
IL-034
Development of bioartificial liver system and potential of stem cells as hepatocyte source
Stress during cell culture processes
Jung-Keug Park 1,2,∗ , Hee-Hoon Yoon 1 , Bo-Young Jung 1 , Ji-Hyun Lee 3 , Suk-Koo Lee 4 , Young-Jin Kim 5 1
Tissue Engineering and Regenerative Medicine Laboratory, Department of Chemical and Biochemical Engineering, Republic of Korea 2 Wave Energy Bioactivation Technology Center, Department of Medical Biotechnology, Dongguk University, Seoul 100-715, Republic of Korea 3 Tissue Engineering Laboratory, Samsung Biomedical Research Institute, Republic of Korea 4 Department of Surgery, Samsung Medical Center, Seoul 135-710, Republic of Korea 5 Cell Therapy Center, Lifecord Inc., Suwon 442-749, Republic of Korea
S115
Joseph Boudrant ∗ , Naziha Barbouche, Nasir Mehmood, Kenza Boulaya, Marie-Franc¸oise Clincke, Eric Olmos, Stéphane Delaunay, Jean-Louis Goergen, Emmanuel Guédon, Annie Marc Laboratory of Chemical Engineering Sciences, UPR CNRS 6811, NancyUniversité, INPL, 2, avenue de la Forêt de Haye, BP 172, 54505 Vandoeuvre-les-Nancy, France E-mail address: [email protected] (J. Boudrant).
Tissue engineering (TE) is an interdisciplinary field that applies medicine, life science, and engineering to develop biological tissues in vitro (Langer and Vacanti, 1993). In Korea, basic researches on TE have grown rapidly last decade. Now, some products are being launched in market and some are under investigational new drug (IND) application. Apart from other TE products, bioartificial liver (BAL) system is a hybrid of biological drug and medical device. To treat fulminant hepatic failure (FHF) patients, various extracorporeal BAL systems have been developed (van de Kerkhove et al., 2004; Park and Lee, 2005). The performance of the BAL depends on the functional activities of the hepatocytes immobilized in the bioreactor system. A packed-bed bioreactor with gel beads has higher surface area to volume ratio and cell capacity than those of a hollow fiber bioreactor. Therefore, the type of bioreactor may be a better alternative for hollow fiber-based BAL systems. In this study, we developed a stable packed-bed bioreactor system that does not induce any damage to the gel beads in the bioreactor. The bioreactor packed with gel bead containing hepatocyte spheroids perfused with down-ward flow of media in which the flow was generated by difference of a media level between the reservoir and outlet chamber placed before and after the bioreactor, respectively. The reactor showed stable liver functions for 12 h, and any damage was not detected at the beads taken from the bioreactor after the long-term operation. And also to acquire human hepatocyte source in the near future, we have checked which stem cells have the hepatic potential and which culture condition can induce them into functional hepatocytes. Now we think that umbilical cord-derived mesenchymal stem cell (UC-MSC) can be a good candidate as human hepatocyte source according to preliminary results.
During cell cultivation, mainly when in suspension, cells generally encounter or have to support various stresses, either physical or chemical. When in liquid medium physical stresses origin can be for example temperature, tensio-active agents, shear stress, light. For cell storage or solid-state fermentation, physical stress can be lack of water. Chemical stresses can be due to variations of pH, of metabolite concentrations, to added chemicals, oxygen, to formed carbon dioxide. The consequences of these stresses can be analyzed in term of viability, of physiological behaviour, for example regarding the produced metabolite, and of genomic (genome-wide expression analyses). This presentation proposes an overview on this large problematic applied to industrial bioprocesses and then focuses on different industrial bacterial fermentations and one animal cell cultivation. The first is producing the amino-acid glutamic acid using a thermoinducible strain of Corynebacterium glutamicum with the effects on the enveloppe and metabolism. The second concerns one filamentous bacteria (Streptomyces prystinaespiralis) producing antibiotics. The third is producing lactic acid with Lactobacillus casei and the study concerns the influence of agitation on the global productivity. The concerned animal cell will be one CHO strain and the study is about the effect of hydrodynamic and medium composition stresses on its global physiological response. This could be also applied to yeast (Saccharomyces cerevisiae) and microalgae cultivations. Moreover, because of large size bioreactors, heterogeneity problems may occur, particularly for temperature or hydrodynamic stress. So, for a serious optimization of the corresponding bioprocesses, the stress distributions in time and space should be seriously quantified. To do this, the local characterization of stresses is expected to bring a more detailed information (quantification, stress duration) than a global one. This implies the use of sophisticated tools (CFP, PIV or LDV). This illustrates that this way of analyzing bioprocesses and their performances is not an easy task, but it could gain more and more importance in the future.
References
doi:10.1016/j.jbiotec.2008.07.371
E-mail address: [email protected] (J.-K. Park).
Langer, R., Vacanti, J.P., 1993. Tissue engineering. Science 260, 920–926. Park, J.K., Lee, D.H., 2005. Bioartificial liver systems; current status and future perspective. J. Biosci. Bioeng. 99, 311–319. van de Kerkhove, M.P., Hoekstra, R., Chamuleau, R.A., van Gulik, T.M., 2004. Clinical application of bioartificial liver support systems. Ann. Surg. 240, 216–230.
doi:10.1016/j.jbiotec.2008.07.370
IL-036 Design, prototyping and operation of disposable bioreactor systems for stem cell cultivation and tissue formation Uwe Marx ∗ , Christian Demmler, Annika Lubitz, Christoph Giese ProBioGen Ag, Goethestrasse, 54 13086 Berlin, Germany E-mail address: [email protected] (U. Marx). Significant progress has been achieved last 20 years in the area of in vitro tissue culture technologies (Marx, 2007). A number of human organ properties could be successfully emulated in vitro (Griffith and Swartz, 2006). Established human skin equivalents, for example, are featuring the evaluation of dermal transfer,