Lactate metabolism shift in CHO cell culture: the role of mitochondrial oxidative activity

被引:146
|
作者
Zagari, Francesca [1 ,2 ]
Jordan, Martin [1 ]
Stettler, Matthieu [1 ]
Broly, Herve [1 ]
Wurm, Florian M. [2 ]
机构
[1] Merck Serono SA, Biotech Proc Sci Grp, CH-1804 Corsier Sur Vevey, Switzerland
[2] Ecole Polytech Fed Lausanne, Lab Cellular Biotechnol, CH-1015 Lausanne, Switzerland
关键词
LACTIC-ACID FORMATION; PYRUVATE-CARBOXYLASE; AEROBIC GLYCOLYSIS; IMPROVEMENT; EXPRESSION; GLUCOSE; REDUCTION; PROTEIN;
D O I
10.1016/j.nbt.2012.05.021
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Lactate production is monitored in industrial processes as a crucial metabolite for cultured mammalian cells. Typically lactate is strongly produced during the exponential growth phase, while its net consumption is frequently observed when cells enter into the stationary phase. Such a metabolic shift is desirable because it seems to favor optimal process performance. However, this shift is neither generic nor can it be easily controlled, as the mechanisms modulating lactate production/consumption in cell culture are still under investigation. In this study different lactate profiles were observed in a chemically defined medium for the parental CHO-S cells and a non-recombinant subclone. The initial lactate production phase, which is typical for fast growing cells, was similar for both cell lines. After glutamine depletion the situation changed: the parental cell line promptly switched to net lactate consumption, whereas the subclone continued to produce lactate until glucose was depleted as well. We speculated that the extra lactate production would be ascribed to a different mitochondrial oxidative capacity in the subclone. Therefore, the mitochondrial membrane potential and oxygen consumption were measured for both cell lines. Indeed, a correlation between high lactate production and a reduced oxidative metabolism was found. Interestingly, this particular metabolic phenotype was also strongly influenced by the medium composition: both cell lines underwent a switch to lactate consumption when cultivated in a second medium, while a third one promoted continuous lactate production even for the parental CHO cells. Again, the correlation between lactate profile and oxidative metabolism was confirmed, pointing to a central role of mitochondria on lactate metabolism.
引用
收藏
页码:238 / 245
页数:8
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