Abstract
Equipment design is frequently recognized as a key component in the success of GMP biologics manufacturing, but is not always implemented with full appreciation of the processing implications. In the case of mammalian cell culture, there are some recognized issues and risks that develop when transitioning to a large scale of operation. The developing demand for cell culture production capacity in the biopharmaceutical industry has led to a progressive increase in the scale of operation in the last decade. This review will provide a high level summary of the documented process difficulties unique to serum-free large scale (LS) cell culture, analyze the engineering constraints typical of these processes, and suggest some practical equipment design considerations to enhance the productivity, reliability and operability of such systems under GMP manufacturing conditions. A systems approach will be used to establish a good LS bioreactor design practice, providing a discussion on gas distribution, agitation, vessel design, SIP/CIP and control issues.
Similar content being viewed by others
References
American Society of Mechanical Engineers 2002. ASME BPE-2002. http: / /www.asmeny.org/ catalog /html/ catgcs.htm
Aunins J.G. and Henzler H.-J. 1993. Aeration in cell culture bioreactors. In: Stephanopoulos G., Rehm H.-J., Reed G., Puhler A. and Stadler P. (eds), Biotechnology Vol. 3. 2nd edn. VCH Verlag GmbH, Weinheim/D, pp. 219–281.
Chalmers J.L. 1994. Cells and bubbles in sparged bioreactors. Cytotechnology 15: 311–320.
Chisti Y. 1993. Animal cell culture in stirred bioreactors: Observations on scale-up. Bioproc Eng 9: 191–196.
Croughan M.S., Hammel J.F. and Wang D.I.C. 1987. Hydrodynamic effects on animal cells grown in microcarrier cultures. Biotechnol Bioeng 29: 130–141.
Croughan M.S., Sayre E.S. and Wang D.I.C. 1989. Viscous reduction of turbulent damage in animal cell culture. Biotechnol Bioeng 33: 862–872.
DeZengotita V., Kimura R. and Miller W. 1998. Effects of CO2 and osmolality on hybridoma cells: growth, metabolism and monoclonal antibody production. Cytotechnology 28: 213–227.
Enfors S.O., Jahic M., Rozkov A., Xu B., Hecker M., Jurgen B. et al. 2001. Physiological responses to mixing in large scale bioreactors. J Biotechnol 85: 175–185.
Garnier A., Voyer R., Tom R., Perret S., Jardin B. and Kamen A. 1996. Dissolved carbon dioxide accumulation in a large scale and high density production of TGFβ receptor with baculovirus infected Sf-9 cells. Cytotechnology 22: 53–63.
Glaser V. 2000. Fermenters and bioreactors for bioprocessing. Genetic Engineering News. Vol. 20, No. 14, pp. 13, 15, 81.
Gray D.R., Chen S., Howarth W., Inlow D. and Majorella B.L. 1996. CO2 in large-scale and high-density CHO cell perfusion culture. Cytotechnology 22: 65–78.
Handa A., Emery A.N. and Spier R.E. 1987. On the evaluation of gas-liquid interfacial effects on hybridoma viability in bubble column bioreactors. Dev Biol Standard 66: 241–253.
Jöbses I., Martens D. and Tramper J. 1990. Lethal events during gas sparging in animal cell culture. Biotechnol Bioeng 37: 484–490.
Kimura R. and Miller W. 1996. Effects of elevated pCO2 and/ or osmolality on the growth and recombinant tPA production of CHO cells. Biotechnol Bioeng 52: 152–160.
Kioukia N., Nienow A.W., Al-Rubeai M. and Emery A.N. 1996. Influence of agitation and sparging on the growth rate and infection of insect cells in bioreactors and a comparison with hybridoma culture. Biotechnol Prog 12: 779–785.
Kunas K.T. and Papoutsakis E.T. 1990. Damage mechanisms of suspended animal cells in agitated bioreactors with and without bubble entrainment. Biotechnol Bioeng 36: 476–483.
Lehmann J., Vorlop J. and Büntemayer H. 1988. Bubble-free reactors and their development for continuous culture with cell recycle. In: Spier R.E. and Griffiths J.B. (eds), Animal Cell Biotechnology Vol. 3. Academic Press Ltd., London, UK, pp. 221–237.
Leist C.H., Meyer H.P. and Fiechter A. 1990. Potential and problems of animal cells in suspension culture. J Biotechnol 15: 1–46.
McQueen A., Meilhoc E. and Bailey J.E. 1987. Flow effectson the viability and lysis of suspended mammalian cells. Biotechnol Bioeng 32: 1001–1014.
Mitchell-Logean C. and Murhammer D.W. 1997. Bioreactor headspace purging reduces dissolved carbon dioxide accumulation in insect cell cultures and enhances cell growth. Biotechnol Prog 13: 875–877.
Morrow J.K. 2002. Economics of antibody production. Genetic Engineering News. Vol. 22, No. 7.
Murhammer D.W. and Goochee C.F. 1990. Sparged animal cell bioreactors: mechanism of cell damage and Pluronic F-68 protection. Biotechnol Prog 6: 391–397.
Nelson K.L. 1988. Industrial scale mammalian cell culture, part II: design and scale-up, Biopharm. Vol. 1, No. 3, pp. 34–41, 50.
Nienow A.W. 1997. On impeller circulation and mixing effectiveness in the turbulent flow regime. Chem Eng Sci 52: 2557–2565.
Nienow A.W., Langheinrich C., Stevenson N.C., Emery A.N., Clayton T.M. and Slater N.K.H. 1996. Homogenization and oxygen transfer rates in large agitated and sparged animal cell bioreactors: Some implications for growth and production. Cytotechnology 22: 87–94.
Ozturk S.S. 1996. Engineering challenges in high density cell culture systems. Cytotechnology 22: 9–13.
Pattison R.N., Swamy J., Mendenhall B., Hwang C. and Frohlich B.T. 2000. Measurement and control of dissolved carbon dioxide in mammalian cell culture processes using an in situ fiber optic chemical sensor. Biotechnol Prog 16: 769–774.
Perry R.H. and Green D.W. 1997. Perry's Chemical Engineers' Handbook. 7th edn. McGraw-Hill, Sec. 18, pp 10.
Qi H., Jovanoic G., Michaels J. and Konstantinov K. 2001. The art and science of micro-sparging in high-density perfusion cultures of animal cells. In: Lindner-Olssen E., Chatzissavidou N. and Lullau E. (eds), Animal Cell Technology: From Target to Market. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp. 412–415.
Schoengerr I., Stapp T. and Ryll T. 2000. A comparison of different methods to determine the end of exponential growth in CHO cell cultures for optimization of scale-up. Biotechnol Prog 16: 815–821.
Sen A., Kallos M.S. and Behie L.A. 2001. Effects of hydrodynamics on cultures of mammalian neural stem cell aggregates in suspension bioreactors. Ind Eng Chem Res 40: 5350–5357.
Spokane R.B., Pitts G.E., Wu P., Cordonnier M.J. and Ordaz D. 1999. An optical sensor for the in situ monitoring of dissolved carbon dioxide in Microbial Fermentations (Abstract). American Chemical Society National Meeting, March 21–25. Biochemical Technology.
Taticek R., Petersen S., Konstantinov K. and Naveh D. 1998. Effect of dissolved carbon dioxide and bicarbonate on mammalian cell metabolism and recombinant protein productivity in high-density perfusion culture. Presented at Cell Culture Engineering Conference VI, San Diego, CA, USA.
Varley J. and Birch J. 1999. Reactor design for large scale suspension animal cell culture. Cytotechnology 29: 177–205.
Vorlop J. and Lehmann J. 1989. Oxygen transfer and carrier mixing in large scale membrane stirred cell culture reactors. In: Spier R.E., Griffiths J.B., Stephenne J. and Crooy P.J., Advances in Animal Cell Biology and Technology for Bioprocesses. Butterworths, Svenoaks, UK, pp. 366–369.
Woodside S.M., Bowen B.D. and Piret J.M. 1998. Mammalian cell retention devices for stirred perfusion bioreactors. Cytotechnology 28: 163–175.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Marks, D.M. Equipment design considerations for large scale cell culture. Cytotechnology 42, 21–33 (2003). https://doi.org/10.1023/A:1026103405618
Issue Date:
DOI: https://doi.org/10.1023/A:1026103405618