title: Regulation of cell growth and apoptosis through lactate dehydrogenase C over-expression in Chinese hamster ovary cells.
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.
authors: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.
authors: Tuo
authors: T
authors: Fu
authors: State Key Laboratory of Antibody Medicine and Targeted Therapy, Shanghai, 201203, China.
authors: Cunchao
authors: C
authors: Zhang
authors: Institutes of Biology and Medical Sciences, Soochow University, Suzhou, 215123, China.
authors: Yu
authors: Y
authors: Jing
authors: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.
authors: Cheng
authors: C
authors: Jiang
authors: State Key Laboratory of Antibody Medicine and Targeted Therapy, Shanghai, 201203, China.
authors: Zhenhua
authors: Z
authors: Li
authors: State Key Laboratory of Antibody Medicine and Targeted Therapy, Shanghai, 201203, China.
authors: Shengyu
authors: S
authors: Wang
authors: State Key Laboratory of Antibody Medicine and Targeted Therapy, Shanghai, 201203, China.
authors: Kai
authors: K
authors: Ma
authors: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Targeted Therapy, Shanghai, 201203, China.
authors: Dapeng
authors: D
authors: Zhang
authors: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Targeted Therapy, Shanghai, 201203, China.
authors: Sheng
authors: S
authors: Hou
authors: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Targeted Therapy, Shanghai, 201203, China.
authors: Jianxin
authors: J
authors: Dai
authors: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China.|State Key Laboratory of Antibody Medicine and Targeted Therapy, Shanghai, 201203, China.
authors: Geng
authors: G
authors: Kou
authors: International Joint Cancer Institute, The Second Military Medical University, Shanghai, 200433, China. hwang_smmu@163.com.|State Key Laboratory of Antibody Medicine and Targeted Therapy, Shanghai, 201203, China. hwang_smmu@163.com.
authors: Hao
authors: H
authors: Wang
mesh_terms: D000818:Animals; D017209:Apoptosis; D061249:Batch Cell Culture Techniques; D016466:CHO Cells; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D053179:Caspase 7 / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation; D002470:Cell Survival; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry; D005786:Gene Expression Regulation; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial; D019289:Pyruvic Acid / Q000737:chemistry; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D019354:Sodium Lactate / Q000737:chemistry
chemical_list: D003470:Culture Media; D007527:Isoenzymes; D011994:Recombinant Proteins; D019289:Pyruvic Acid; D007770:L-Lactate Dehydrogenase; C409673:lactate dehydrogenase C4; D053148:Caspase 3; D053179:Caspase 7; D019354:Sodium Lactate
keywords: Apoptosis; Chinese hamster ovary cells; Lactate; Lactate dehydrogenase C; Sodium pyruvate