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. 2001 Oct;37(2):93–105. doi: 10.1023/A:1019908310300

Amino acids metabolism by VO 208 hybridoma cells: some aspects of the culture process and medium composition influence

A Martial-Gros 1, JL Goergen 1, JM Engasser 1, A Marc 1
PMCID: PMC3449694  PMID: 19002906

Abstract

In the present study an approach has been developed in order to examine the consequence of essential and non essential amino acid supplementation on VO208 hybridoma cells behaviour. The effect of amino acid enrichment has been studied taking into account the culture process, i.e., batch or continuous culture mode and the medium composition, i.e., a home made serum-free medium or a serum containing one. A group of 4 amino acids, i.e., Ser, Pro, Gly and Arg presented atypical evolution pattern of their extracellular concentration depending on the type of the medium and on the culture mode. Some amino acids were probably involved in the limitation of the cellular proliferation. Met was one of the amino acids that appears to may have been at limiting concentration in all cases. In continuous culture mode, an enrichment of amino acids resulted in a rapid improvement of the viable cell density in both media, with or without the presence of serum. For most amino acids, supplementation during continuous culture induced an increase of the amino acid uptake rate. A comparative analysis of amino acids utilisation, depending on the culture conditions studied in the present study, has been performed in order to propose an overall picture of amino acids metabolism by VO 208 Hybridoma cell line.

Keywords: Amino acids, Batch culture, Continuous culture, Hybridoma, Kinetics, Medium composition, Metabolism, Serum

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References

  1. Baydoun A.R., Emery P.W., Pearson J.D., Mann G.E. Substrate-dependent regulation of intracellular amino acid concentrations in cultured bovine aortic endothelial cells. Biochem. Biophys. Res. Commun. 1990;173:940–948. doi: 10.1016/S0006-291X(05)80876-9. [DOI] [PubMed] [Google Scholar]
  2. Bibila A.T., Ranucci S.C., Glazomitsky K., Buckland C.B., Aunins J.G. Monoclonal antibody process development using medium concentrates. Biotechnol. Prog. 1994;10:87–96. doi: 10.1021/bp00025a011. [DOI] [PubMed] [Google Scholar]
  3. Borth N., Heider R., Assadian A., Katinger H. Growth and production kinetics of human-mouse and mouse hybridoma cells at reduced temperature and serum content. J. Biotechnol. 1992;25:319–331. doi: 10.1016/0168-1656(92)90164-5. [DOI] [PubMed] [Google Scholar]
  4. Blasey H.D., Winzer V. Low protein serum-free medium for antibody production in stirred reactors. Biotechnol. Lett. 1989;11:455–460. doi: 10.1007/BF01026641. [DOI] [Google Scholar]
  5. Butler M., Imamura T., Thomas J., Thilly W.G. High yields from microcarrier cultures by medium perfusion. J. Cell Sci. 1983;61:351–363. doi: 10.1242/jcs.61.1.351. [DOI] [PubMed] [Google Scholar]
  6. Duval D., Geahel I., Dufau A.F. and Hache J. 1989. Effect of amino acids on the growth and productivity of hybridoma cell cultures. In: Spier R.E., Griffiths J.B., Stephenne J. and Crooy P.J. (eds), Advances in animal cell biology and technology for bioprocesses. Butterworths, pp. 257–259.
  7. Franek F., Sramkova C. Apoptosis and nutrition: in-volvement of amino acid transport system in repression of hybridoma cell death. Cytotechnology. 1995;18:113–117. doi: 10.1007/BF00744326. [DOI] [PubMed] [Google Scholar]
  8. Geaugey V., Duval D., Geahel I., Marc A., Engasser J.M. Influence of amino acids on hybridoma cell viability and antibody secretion. Cytotechnology. 1989;2:119–129. doi: 10.1007/BF00386144. [DOI] [PubMed] [Google Scholar]
  9. Gelot M.A., Jendel C., Belleville F., Nabet P., Penin P., Cuny G. Acides aminés du liquide céphalorachidien dans la maladie d'Alzheimer. Ann. Biol. Clin. 1990;48:182–184. [PubMed] [Google Scholar]
  10. Gottesman M.M. Chinese Hamster Ovary Cells. Methods Enzymol. 1982;151:3–8. doi: 10.1016/S0076-6879(87)51004-7. [DOI] [PubMed] [Google Scholar]
  11. Haüssinger D., Lang F., Kilberg M.S. Amino acid transport, cell volume and regulation of cell growth. In: Kilberg M.S., Haüssinger D., editors. Mammalian amino acid transport. New York: Plenum Press; 1992. pp. 113–130. [Google Scholar]
  12. Martial A., Dousset B., Dardenne M., Engasser J.M., Nabet P., Marc A. Insulin utilization and kinetic effect on hybridoma metabolism in batch and continuous cultures. J. Biotechnol. 1994;34:195–203. doi: 10.1016/0168-1656(94)90089-2. [DOI] [PubMed] [Google Scholar]
  13. Miller W.M., Wilke C., Blanch H.W. A kinetic analysis of hybridoma growth and metabolism in batch and continuous suspension culture: effect of nutrient concentration, dilution rate and pH. Biotechnol. Bioeng. 1988;32:947–965. doi: 10.1002/bit.260320803. [DOI] [PubMed] [Google Scholar]
  14. Miller W.M., Wilke C.R., Blanch H.W. Transient responses of hybridoma cells to nutrient addition in continuous culture. I: Glucose pulse and step changes. Biotechnol. Bioeng. 1989;33:477–486. doi: 10.1002/bit.260330413. [DOI] [PubMed] [Google Scholar]
  15. Miller W.M., Wilke C.R., Blanch H.W. Transient responses of hybridoma cells to nutrient addition in continuous culture. II: Glutamine pulse and step changes. Biotechnol. Bioeng. 1989;33:487–499. doi: 10.1002/bit.260330414. [DOI] [PubMed] [Google Scholar]
  16. Morgan J.F., Morton H.J. The nutrition of animal tissus cultivated in vitro. IV: Amino acid requirement of chick em-bryonic heart fibroblasts. J. Biophys. Biochem. Cytol. 1957;3:141. doi: 10.1083/jcb.3.2.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Phang J.M. The regulatory functions of proline and pyrroline-5-carboxilic acid. In: Horecker B.L., Stadtman E.R., editors. Current topics in cellular regulation. New York: Academic Press; 1985. pp. 91–132. [DOI] [PubMed] [Google Scholar]
  18. Pialkowski S.E. Real time digital filters. Anal. Chem. 1988;60:355A–361A. [Google Scholar]
  19. Reddy S., Kenneth D.B., Miller W.M. Determination of antibody content in live versus dead hybridoma cells: analysis of antibody production in osmotically stressed cultures. Biotechnol. Bioeng. 1992;40:947–964. doi: 10.1002/bit.260400811. [DOI] [PubMed] [Google Scholar]
  20. Seewöster T., Lehmann J. Influence of targeted asparagine starvation on extra-and intracellular amino acid pools of cultivated Chinese hamster ovary cells. Appl. Microbiol. Biotechnol. 1995;44:344–350. doi: 10.1007/BF00169927. [DOI] [PubMed] [Google Scholar]
  21. Sukemori S., Sugimura K. Studies on the influence of amino acid imbalance on mammalian cells. J. Anim. Physiol. Anim. Nutr. 1991;66:241–247. doi: 10.1111/j.1439-0396.1991.tb00293.x. [DOI] [Google Scholar]
  22. Wagner A., Villermaux S., Marc A., Engasser J.M., Einsele A. Continuous production of Prourokinase in feed-harvest and perfusion cultures. Biotechnol. Bioeng. 1990;36:623–629. doi: 10.1002/bit.260360610. [DOI] [PubMed] [Google Scholar]
  23. Xie L., Wang D.I.C. High cell density and high monoclonal antibody production through medium design and rational control in bioreactor. Biotechnol. Bioeng. 1996;51:725–729. doi: 10.1002/(SICI)1097-0290(19960920)51:6<725::AID-BIT12>3.0.CO;2-C. [DOI] [PubMed] [Google Scholar]
  24. Zhou W., Chen C.C., Buckland B., Aunins J. Fed-batch culture of recombinant NSO myeloma cells with high mono-clonal antibody production. Biotechnol. Bioeng. 1997;5:783–792. doi: 10.1002/(SICI)1097-0290(19970905)55:5<783::AID-BIT8>3.0.CO;2-7. [DOI] [PubMed] [Google Scholar]

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