Alina Rekena, Dora Livkisa, Dagnija Loca


Advantageous cultivation procedures for the Chinese hamster ovary (CHO) cells are necessary for the productive commercial production of biopharmaceuticals. A main challenge that needs to be addressed during the process development is the differences in each cell line requirements concerning the nutrients and feed strategies in order to achieve the desired growth characteristics. Therefore, within the current research, a naïve high cell density serum free suspension adapted CHO cell line was tested with glucose and glutamine rich feeds in fed-batch Erlenmeyer shake flask cultures. Glucose consumption rate was adjusted to develop the optimal feed strategies. Obtained results indicated that high glucose and l-glutamine feeding did not improve maximum viable cell density compared to the control samples. During the exponential phase, cell proliferation and viability of all feeds showed no statistically significant difference. Instead, the fed-batch processes tested led to statistically significant differences in viable cell density and cell viability during the decline phase, compared to control (batch) culture. The difference between glucose and glutamine feeding was indistinguishable, most probably due to the concentration imbalance with the rest of the nutrients in feed. The overall study presented a method to slow down the decrease in CHO cell proliferation and viability during the decline phase, instead of increasing the maximum cell density at the plateau phase.



biotechnology; cell proliferation; Chinese hamster ovary cells; feeding strategy

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J. Zhu, “Mammalian cell protein expression for biopharmaceutical production,” Biotechnol. Adv., vol. 30, no. 5, pp. 1158–1170, 2012.

F. V. Ritacco, Y. Wu, and A. Khetan, “Cell Culture Media for Recombinant Protein Expression in Chinese Hamster Ovary (CHO) Cells: History, Key Components, and Optimization Strategies,” Biotechnol Prog, vol. 34, no. 6, pp. 1407–1426, 2018.

P. Xu, X. P. Dai, E. Graf, R. Martel, and R. Russell, “Effects of glutamine and asparagine on recombinant antibody production using CHO-GS cell lines,” Biotechnol. Prog., vol. 30, no. 6, pp. 1457–1468, 2014.

J. A. H. Bort, B. Stern, and N. Borth, “CHO-K1 host cells adapted to growth in glutamine-free medium by FACS-assisted evolution,” Biotechnol. J., vol. 5, no. 10, pp. 1090–1097, 2010.

L. E. Quek, S. Dietmair, J. O. Krömer, and L. K. Nielsen, “Metabolic flux analysis in mammalian cell culture,” Metab. Eng., vol. 12, no. 2, pp. 161–171, 2010.

C. A. Wilkens, C. Altamirano, and Z. P. Gerdtzen, “Comparative metabolic analysis of lactate for CHO cells in glucose and galactose,” Biotechnol. Bioprocess Eng., vol. 16, no. 4, pp. 714–724, 2011.

S. Lu, X. Sun, and Y. Zhang, “Insight into metabolism of CHO cells at low glucose concentration on the basis of the determination of intracellular metabolites,” Process Biochem., vol. 40, no. 5, pp. 1917–1921, 2005.

P. Chen and S. W. Harcum, “Effects of amino acid additions on ammonium stressed CHO cells,” J. Biotechnol., vol. 117, no. 3, pp. 277–286, 2005.

A. ‐P Zeng and W. ‐D Deckwer, “Mathematical modeling and analysis of glucose and glutamine utilization and regulation in cultures of continuous mammalian cells,” Biotechnol. Bioeng., vol. 47, no. 3, pp. 334–346, 1995.

H. Wlaschin, Katie F., Wei-Shou, “Fedbatch Culture and Dynamic Nutrient Feeding,” Adv. Biochem. Eng. Biotechnol., vol. 101, pp. 43–74, 2006.

J. Robitaille, J. Chen, and M. Jolicoeur, “A single dynamic metabolic model can describe mAb producing CHO cell batch and fed-batch cultures on different culture media,” PLoS One, vol. 10, no. 9, 2015.

A. R. Costa, M. E. Rodrigues, M. Henriques, R. Oliveira, and J. Azeredo, “Feed optimization in fed-batch culture,” Methods Mol. Biol., vol. 1104, pp. 105–116, 2014.

X. Sun and Y. Zhang, “Glutamine cannot support recombinant CHO cell growth and maintenance in the absence of glucose,” Process Biochem., vol. 39, no. 6, pp. 717–720, 2004.

X. Pan, M. Streefland, C. Dalm, R. H. Wijffels, and D. E. Martens, “Selection of chemically defined media for CHO cell fed-batch culture processes,” Cytotechnology, vol. 69, no. 1, pp. 39–56, 2017.

E. Pacis, N. Vijayasankaran, J. Li, M. Gawlitzek, A. Amanullah, and F. Li, “Systematic approaches to develop chemically defined cell culture feed media,” BioPharm Int., vol. 23, no. 11, pp. 22–32, 2010.

DOI: http://dx.doi.org/10.17770/etr2019vol1.4106


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