Prolongation of murine hybridoma cell survival in stationary batch culture by Bcl-xL expression

被引：0
作者：
Joel R. Charbonneau
Eric R. Gauthier
机构：
[1] Laurentian University,Cellular Biochemistry Research Laboratory, Department of Chemistry and Biochemistry
[2] Laurentian University,Cellular Biochemistry Research Laboratory, Department of Chemistry and Biochemistry
来源：
Cytotechnology | 2000年 / 34卷
关键词：
apoptosis; bcl-xL; cell growth; cell viability; hybridoma; myeloma;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
While the ectopic expression of the anti-apoptoticprotein Bcl-2 has been shown to significantly increaseboth cell viability and antibody production in batchculture, some cell lines are refractory to thesemanipulations. For example, the NS/O and theP3x63Ag8.653 murine myelomas, which express highendogenous levels of the Bcl-2 homologue Bcl-xL, areboth resistant to the anti-apoptotic effect of Bcl-2.This indicates that, in these cells, Bcl-2 and Bcl-xLmay be functionally redundant. In order to define therole which Bcl-xL plays in hybridoma cultures, we usedthe Sp2/0-Ag14 cell line. This murine hybridomaexpresses low levels of Bcl-xL and is highly sensitiveto apoptosis induction by cycloheximide (CHX) and byamino acid depletion. Bcl-xL-transfected Sp2/0-Ag14cells were more resistant than the wild type and theplasmid-containing cells to apoptosis induced by CHXand by glutamine depletion. Moreover, when compared tothe vector-transfected control, Bcl-xL-Sp2/0 cellsexhibited a substantial increase in viability instationary batch culture. Interestingly, Sp2/0-Ag14cells overexpressing Bcl-xL showed a growth behaviourthat was similar to the parent myeloma cell lineP3x63Ag8.653. Our results suggest that Bcl-xLexpression levels are sufficient to account for therelative robustness of some hybridoma cell lines instationary batch cultures.
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页码：131 / 139
页数：8
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共 111 条
[11]  
Shore GC(1992)Fragmented DNA and apoptotic bodies document the programmed way of cell death in hybridoma cultures Cytotechnology 9 117-123
[12]  
Chiou SK(1997)Reinforcing apoptosis-resistance of COS and myeloma cells by transfecting with bcl-2 gene Cytotechnology 25 25-33
[13]  
Tseng CC(1996)Role of bcl-xL in the control of apoptosis in murine myeloma cells Cancer Res 56 1451-1456
[14]  
Rao L(1998)Mitochondria and apoptosis Science 281 1309-1312
[15]  
White E(1989)Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill J Immunol Methods 119 203-210
[16]  
Cotter TG(1995)Overexpression of bcl-2, apoptosis suppressing gene: prolonged viable culture period of hybridoma and enhanced antibody production Biotechnol Bioeng 48 118-122
[17]  
Al-Rubeai M(1998)Overcoming apoptosis: new methods for improving protein-expression systems Trends Biotechnol 16 88-95
[18]  
Dickson AJ(2000)Overexpression of Bcl-2 family members enhances survival of mammalian cells in response to various culture insults Biotechnol Bioeng 67 555-564
[19]  
Fadeel B(1999)Dose-dependent reduction of apoptosis in nutrient-limited cultures of NS/0 myeloma cells transfected with the E1B-19K adenoviral gene Biotechnol Bioeng 63 516-528
[20]  
Zhivotovsky B(1994)Induction of apoptosis in nutrientdeprived cultures of hybridoma and myeloma cells Biotechnol Bioeng 44 1140-1154
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