Abstract
Chinese Hamster Ovary (CHO) cells are used for the production of therapeutic proteins. This work examines improving passaging growth rate of two CHO clones. Growth rates were significantly improved for both clones with supplementation of the nucleosides cytidine, hypoxanthine, uridine, and thymidine to the culturing media at the optimal concentration of 100 µM of each nucleoside. We investigated supplementing the same combination of nucleosides to seed bioreactors and production fed batch bioreactors. In the seed bioreactors, growth rate and harvest density were improved. However, in the production fed batch bioreactors, no improvements in growth rate or peak viable cell density were observed. Cell cycle analysis of the passaging cells provides evidence that nucleosides can affect the cell cycle. It is not clear from our work how the nucleosides impact the cell cycle regulatory pathways. Overall, nucleoside supplementation in cell culture media is an effective approach for improving growth rate in passaging and seed bioreactors of certain CHO cells.
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Acknowledgements
The authors would like to acknowledge Paulena Lieske for her help setting up and running the Fortessa® for the cell cycle analysis. The authors would like to acknowledge Dawn Erikson-Stapleton, Matt Gagnon, and Greg Hiller for their comments and suggestions during manuscript preparation.
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Morrison, C., Bandara, K., Wang, W. et al. Improvement of growth rates through nucleoside media supplementation of CHO clones. Cytotechnology 71, 733–742 (2019). https://doi.org/10.1007/s10616-019-00319-0
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DOI: https://doi.org/10.1007/s10616-019-00319-0