Abstract
We investigated the relationship between the specific production rate and the concentration of the toxic drug methotrexate (MTX), inhibitory to nucleic acid synthesis in the medium, during the construction of recombinant CHO cells. The results of selection of MTX tolerant cells when the MTX concentration was increased under 10 different stepwise conditions showed that the number of days required to obtain cells resistant to 1000 nM MTX depended on the selection conditions. Tolerant cells were obtained most rapidly at an MTX concentration of 10nM. With increasing MTX concentration, the specific production rate increased, while the specific growth rate gradually decreased. In the case of a gradual increase in the MTX concentration, the specific production rate reached a higher level. We also investigated the stability of the specific production rate during cultivation over a long period with or without MTX. In long-term cultivation, the specific production rate decreased, while the specific growth rate remained at the same level. The specific production rate in a culture without MTX decreased more rapidly than that in a culture with MTX.
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Omasa T., Ishimoto M., Higashiyama K., Shioya S., and Suga K. (1992) The enhancement of specific antibody production rate in glucose — and glutamine — controlled fed-batch culture’ Cytotechnology, 8, 75–84
Kuriki K., Sonta S., and Murata K. (1993) ‘Flow kaiyotype analysis and sorting of the Chinese hamster chromosomes: Comparing the effects of the isolation buffers’ J. Clin. Lab. Anal, 7, 119–122.
Schimke R. T., Brown P. C, and Kaufman R. J. (1982) ‘Gene amplification and drug resistance in mammalian cells’ Natl. Cancer Inst. Monogr. 60, 79–86.
Ruiz J. C. and Wahl G. M. (1990) ‘Chromosomal destabilization during gene amplification’ Mol. Cell. Biol. 10, 3056–3066
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© 1998 Springer Science+Business Media Dordrecht
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Omasa, T., Itami, S., Kameoka, D., Katakura, Y., Suga, Ki. (1998). Selection and Stability for a Recombinant CHO Cell Line Expressing Human GM-CSF in Gene Amplification. In: Nagai, K., Wachi, M. (eds) Animal Cell Technology: Basic & Applied Aspects. Animal Cell Technology: Basic & Applied Aspects, vol 9. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-5161-0_6
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DOI: https://doi.org/10.1007/978-94-011-5161-0_6
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-6170-4
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