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Bcl-2 over-expression reduced the serum dependency and improved the nutrient metabolism in a NS0 cells culture

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Abstract

The over-expression of Bcl-2 has greatly improved the culture period, specific growth rate, and maximum viable cell density of NS0 cells culture under low serum condition. Further analysis of these data suggests that a saturation model of the Monod type can be used to represent the relationships of specific growth rate and initial serum concentration. The μmax andK s for the Bcl-2 cell line is 0.927 day−1 and 0.947% (v/v) respectively, which are 21% greater and 7% lower respectively than its control counterpart. Study on the amino acid supplementation revealed that Bcl-2 cell lines possess greater improvement in the specific growth rate and maximum viable cell density compared to the control cell lines. A further increase in the amino acid supplementation has resulted a 17% decrease in specific growth rate and no improvement in maximum viable cell density in the control culture. However, the Bcl-2 cell line exhibited a better growth characteristic in this culture condition compared to that of control cell lines. The higher specific growth rate and maximum viable cell density of the Bcl-2 cell line in medium fortified with serum and MEM EAA suggested a more efficient nutrient metabolism compared to that in the control cell line. The low serum and amino acid utilisation rate and the higher cell yield may prove to be important in the development of serum/protein free culture.

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References

  1. Renner, W. A., K. H. Lee, V. Hatzimanikatis, J. E. Bailey, and H. M. Eppenberger (1995) Recombinant cyclin-E expression activates proliferation and obviates surface attachment of chinese hamster ovary (CHO) cells in protein-free medium.Biotechnol. Bioeng. 47: 476–482.

    Article  CAS  Google Scholar 

  2. Zanghi, I. A., M. Fussenegger, and J. E. Bailey (1999) Serum protects protein-free competent Chinese hamster ovary cells against apoptosis induced by nutrient deprivation in batch culture.Biotechnol. Bioeng. 64: 108–119.

    Article  CAS  Google Scholar 

  3. Tey, B. T., R. P. Singh, and M. Al-Rubeai (2001) Programmed cell death: An overview of apoptosis in cell culture.Asia Pac. J. Mol. Biol. Biotechnol. 9: 1–28.

    Google Scholar 

  4. Itoh, Y., H. Ueda, and E. Suzuki (1995) Over-expression of Bcl-2. apoptosis suppressing gene-prolonged viable culture period of hybridoma and enhanced antibody-production.Biotechnol. Bioeng. 48: 118–122.

    Article  CAS  Google Scholar 

  5. Simpson, N. H., A. E. Milner, and M. Al-Rubeai (1997) Prevention of hybridoma cell death by Bcl-2 during suboptimal culture conditions.Biotechnol. Bioeng. 54: 1–16.

    Article  CAS  Google Scholar 

  6. Tey, B. T., R. P. Singh, L. Piredda, M. Piacentini, and M. Al-Rubeai (2000) Bcl-2 mediated suppression of apoptosis in myeloma NS0 cultures.J. Biotechnol. 79: 147–159.

    Article  CAS  Google Scholar 

  7. Goswami, J., A. J. Sinskey, H. Steller, G. N. Stephanopoulos, and D. I. C. Wang (1999) Apoptosis in batch cultures of Chinese hamster ovary cells.Biotechnol. Bioeng. 62: 632–640.

    Article  CAS  Google Scholar 

  8. Mastrangelo, A. J., J. M. Hardwick, F. Bex, and M. J. Betenbaugh (2000) Part I. Bcl-2 and Bcl-x(L) limit apoptosis upon infection with alphavirus vectors.Biotechnol. Bioeng. 67: 544–554.

    Article  CAS  Google Scholar 

  9. Tey, B. T., R. P. Singh, L. Piredda, M. Piacentini, and M. Al-Rubeai (2000) Influence of Bcl-2 on cell death during cultivation of a Chinese hamster ovary cell line expressing a chimeric antibody.Biotechnol. Bioeng. 68: 31–43.

    Article  CAS  Google Scholar 

  10. Singh, R. P., M. Al-Rubeai, C. D. Gregory, and A. N. Emery (1994) Cell-death in bioreactors: A role for apoptosis.Biotechnol. Bioeng. 44: 720–726.

    Article  CAS  Google Scholar 

  11. Mercille, S. and B. Massie (1994) Induction of apoptosis in nutrient-deprived cultures of hybridoma and myeloma cells.Biotechnol. Bioeng. 44: 1140–1154.

    Article  CAS  Google Scholar 

  12. Simpson, N. H., R. P. Singh, A. Perani, C. Goldenzon, and M. Al-Rubeai (1998) In hybridoma cultures, deprivation of any single amino acid leads to apoptotic death, which is suppressed by the expression of thebcl-2 gene.Biotechnol. Bioeng. 59: 90–98.

    Article  CAS  Google Scholar 

  13. Bebbington, C. R. and C. G. Hentschel (1987) The use of vectors based on gene amplication for the expression of cloned genes in mammalian cells. pp. 163–187. In: D. M. Glover (ed.).DNA Cloning: A Practical Approach Vol. 3. IRL, Oxford, UK.

    Google Scholar 

  14. Leelavatcharamas, V. (1997)Growth, Gamma Interferon Production and Cell Cycle in Batch, Continuous and Perfusion Cultures. Ph.D. Thesis. University of Birmingham, UK.

    Google Scholar 

  15. Dalili, M. and D. F. Ollis (1989) Transient kinetics of hybridoma growth and monoclonal-antibody production in serum-limited cultures.Biotechnol. Bioeng. 33: 984–990.

    Article  CAS  Google Scholar 

  16. Leelavatcharamas, V., A. N. Emery, and M. Al-Rubeai (1994) Growth and interferon-gamma production in batch culture of CHO cells.Cytotechnology 15: 65–71.

    Article  CAS  Google Scholar 

  17. Singh, R. P., A. N. Emery, and M. Al-Rubeai (1996) Enhancement of survivability of mammalian cells by over-expression of the apoptosis-suppressor genebcl-2.Biotechnol. Bioeng. 52: 166–175.

    Article  CAS  Google Scholar 

  18. Fassnacht, D., S. Rossing, F. Franek, M. Al-Rubeai, and R. Portner (1998) Effect of Bcl-2 expression on hybridoma cell growth in serum- supplemented, protein-free and diluted media.Cytotechnology 26: 219–225.

    Article  CAS  Google Scholar 

  19. Franek, F. and J. Dolnikova (1991) Nucleosomes occurring in protein-free hybridoma cell-culture: Evidence for programmed cell-death.FEBS Lett. 284: 285–287.

    Article  CAS  Google Scholar 

  20. Franek, F. (1995) Starvation-induced programmed death of hybridoma cells, prevention by amino-acid mixtures.Biotechnol. Bioeng. 45: 86–90.

    Article  CAS  Google Scholar 

  21. Ishaque, A. and M. Al-Rubeai (1999) Role of Ca, Mg and K ions in determining apoptosis and extent of suppression afforded by bcl-2 during hybridoma cell culture.Apoptosis 4: 335–355.

    Article  CAS  Google Scholar 

  22. Ishaqe, A. and M. Al-Rubeai (2002) Role of vitamins in determining apoptosis and extent of suppression by bcl-2 during hybridoma cell culture.Apoptosis 7: 231–239.

    Article  Google Scholar 

  23. Tey, B. T. and M. Al-Rubeai (2004) Suppression of apoptosis in perfusion culture of Myeloma NS0 cells enhances cell growth but reduces antibody productivity.Apoptosis 9: 843–852.

    Article  CAS  Google Scholar 

  24. Plas, D. R. and C. B. Thompson (2002) Cell metabolism in the regulation of programmed cell death.Trends Endocrin. Met. 13: 74–78.

    Article  CAS  Google Scholar 

  25. Susin, S. A., H. K. Lorenzo, N. Marzo, I. Zamzami, B. E. Snow, G. M. Brothers, J. Mangion, E. Jacotot, P. Costantini, M. Loeffler, N. Larochette, D. R. Goodlett, R. Aebersold, D. P. Siderovski, J. M. Penninger, and G. Kroemer (1999) Molecular characterization of mitochondrial apoptosis-inducing factor.Nature 397: 441–446.

    Article  CAS  Google Scholar 

  26. Al-Rubeai, M. (1998) Apoptosis and cell culture technology.Adv. Biochem. Eng. Biotechnol. 59: 226–249.

    Google Scholar 

  27. Xie, L. Z. and D. I. C. Wang (1996) Energy metabolism and ATP balance in animal cell cultivation using a stolchiometrically based reaction network.Biotechnol. Bioeng. 52: 591–601.

    Article  CAS  Google Scholar 

  28. Rabinovitz, M. (1992) The pleiotypic response to amino-acid deprivation is the result of interactions between components of the glycolysis and protein-synthesis pathways.FEBS Lett. 302: 113–116.

    Article  CAS  Google Scholar 

  29. Tey, B. T. and M. Al-Rubeai (2005) Effect of Bcl-2 over-expression on cell cycle and antibody productivity in chemostat cultures of myeloma NS0 cells.J. Biosci. Bioeng. (In Press).

  30. Castagna, M., C. Shayakul, D. Trotti, V. F. Sacchi, W. R. Harvey, and M. A. Hediger (1997) Molecular characteristics of mammalian and insect amino acid transporters: Implications for amino acid homeostasis.J. Exp. Biol. 200: 269–286.

    CAS  Google Scholar 

  31. Franek, F. and K. ChladkovaSramkova (1995) Apoptosis and nutrition: Involvement of amino acid transport system in repression of hybridoma cell death.Cytotechnology 18: 113–117.

    Article  CAS  Google Scholar 

  32. Franek, F. and K. Sramkova (1996) Cell suicide in starving hybridoma culture: Survival-signal effect of some amino acids.Cytotechnology 21: 81–89.

    Article  CAS  Google Scholar 

  33. Franek, F. and K. Sramkova (1996) Protection of B lymphocyte hybridoma against starvation-induced apoptosis: Survival-signal role of some amino acids.Immunol. Lett. 52: 139–144.

    Article  CAS  Google Scholar 

  34. Kurita, T. and H. Namiki (1994) Apoptotic cell-death induced by serum and its prevention by thiols.J. Cell. Physiol. 161: 63–70.

    Article  CAS  Google Scholar 

  35. Ratan, R. R. and J. M. Baraban (1995) Apoptotic death in anin vitro model of neuronal oxidative stress.Clin. Exp. Pharmacol. Physiol. 22: 309–310.

    Article  CAS  Google Scholar 

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Tey, B.T., Al-Rubeai, M. Bcl-2 over-expression reduced the serum dependency and improved the nutrient metabolism in a NS0 cells culture. Biotechnol. Bioprocess Eng. 10, 254–261 (2005). https://doi.org/10.1007/BF02932022

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