Bcl-x(L) overexpression delays the onset of autophagy and apoptosis in hyperosmotic recombinant Chinese hamster ovary cell cultures |
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Authors: | Han Young Kue Ha Tae Kwang Kim Yeon-Gu Lee Gyun Min |
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Affiliation: | Department of Biological Sciences, Graduate School of Nanoscience & Technology (WCU), KAIST, 373-1 Kusong-Dong, Yusong-gu, Daejon 305-701, Republic of Korea |
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Abstract: | Hyperosmolality in recombinant Chinese hamster ovary (rCHO) cell cultures induces autophagy and apoptosis. To investigate the effect of Bcl-xL overexpression on autophagy and apoptosis in hyperosmotic rCHO cell cultures, an erythropoietin (EPO)-producing rCHO cell line with regulated Bcl-xL overexpression was subjected to hyperosmolality resulting from NaCl addition in a batch culture and nutrient supplementation in a fed-batch culture. In the batch culture, Bcl-xL overexpression suppressed apoptosis, as evidenced by a decreased amount of cleaved caspase-7 and PARP. Concurrently, Bcl-xL overexpression also delayed autophagy, as indicated by reduced LC3 conversion, from LC3-I to LC3-II. As a result, the cell viability and EPO production were improved by Bcl-xL overexpression. In the fed-batch culture, the simultaneous application of Bcl-xL overexpression and nutrient feeding increased the culture longevity and maximum EPO concentration. Taken together, Bcl-xL overexpression delayed autophagy and apoptosis in hyperosmotic rCHO cell cultures, resulting in increased EPO production. |
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Keywords: | Bcl-xL Autophagy Apoptosis rCHO cells Hyperosmolality Fed-batch culture |
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