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Regulation of cell growth and apoptosis through lactate dehydrogenase C over-expression in Chinese hamster ovary cells

  • Applied genetics and molecular biotechnology
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Abstract

Lactate has long been credited as a by-product, which jeopardizes cell growth and productivity when accumulated over a certain concentration during the manufacturing process of therapeutic recombinant proteins by Chinese hamster ovary (CHO) cells. A number of efforts to decrease the lactate concentration have been developed; however, the accumulation of lactate is still a critical issue by the late stage of fed-batch culture. Therefore, a lactate-tolerant cell line was developed through over-expression of lactate dehydrogenase C (LDH-C). In fed-batch culture, sodium lactate or sodium pyruvate was supplemented into the culture medium to simulate the environment of lactate accumulation, and LDH-C over-expression increased the highest viable cell density by over 30 and 50 %, respectively, on day 5, meanwhile the viability was also improved significantly since day 5 compared with that of the control. The percentages of cells suffering early and late apoptosis decreased by 3.2 to 12.5 and 2.0 to 4.3 %, respectively, from day 6 onwards in the fed-batch culture when 40 mM sodium pyruvate was added compared to the control. The results were confirmed by mitochondrial membrane potential assay. In addition, the expression of cleaved caspases 3 and 7 decreased in cells over-expressing LDH-C, suggesting the mitochondrial pathway was involved in the LDH-C regulated anti-apoptosis. In conclusion, a novel cell line with higher lactate tolerance, lowered lactate production, and alleviated apoptosis response was developed by over-expression of LDH-C, which may potentially represent an efficient and labor-saving approach in generating recombinant proteins.

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Acknowledgments

This work was supported in part by grants from the National Natural Science Foundation of China (81330061), Special Project for Infection Disease and New Drug Development, Shanghai Commission of Science and Technology, and Shanghai Municipal Commission for Shanghai Leading Academic Project (B905).

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Authors and Affiliations

  1. International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China

    Tuo Fu, Cheng Jiang, Dapeng Zhang, Sheng Hou, Jianxin Dai, Geng Kou & Hao Wang

  2. State Key Laboratory of Antibody Medicine and Targeted Therapy, Shanghai, 201203, China

    Cunchao Zhang, Zhenhua Li, Shengyu Wang, Kai Ma, Dapeng Zhang, Sheng Hou, Jianxin Dai, Geng Kou & Hao Wang

  3. Institutes of Biology and Medical Sciences, Soochow University, Suzhou, 215123, China

    Yu Jing

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  1. Tuo Fu
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  2. Cunchao Zhang
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  3. Yu Jing
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  4. Cheng Jiang
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Correspondence to Hao Wang.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Tuo Fu and Cunchao Zhang contributed equally to this study.

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Fu, T., Zhang, C., Jing, Y. et al. Regulation of cell growth and apoptosis through lactate dehydrogenase C over-expression in Chinese hamster ovary cells. Appl Microbiol Biotechnol 100, 5007–5016 (2016). https://doi.org/10.1007/s00253-016-7348-4

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  • DOI: https://doi.org/10.1007/s00253-016-7348-4

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