Effect of glutamine limitation on the death of attached Chinese hamster ovary cells.
The effect of glutamine depletion on the death of attached Chinese hamster ovary (CHO) cells was investigated. Experiments were performed using an anchorage dependent CHO cell line expressing gamma-IFN and a second cell line obtained by transfection of that cell line with the human bcl-2 (hbcl-2). Either cell line could grow in media devoid of glutamine with minimal cell death due to endogenous glutamine synthetase activity that allowed cells to synthesize glutamine from glutamic acid in the medium. However, compared to control cultures in glutamine-containing media, the cell growth rate in glutamine-free media was slower with an increased fraction of cells distributed in the G0/G1 phase. The slower rate of cell cycling apparently protected the cells from entering apoptosis when they were stimulated to proliferate in an environment devoid of other protective factors, such as serum or over-expressed hbcl-2. The depletion of both glutamine and glutamic acid did cause cell death, which could be mitigated by hbcl-2 over-expression.
Sanfeliu
A
A
Department of Chemical Engineering and Biotechnology Process Engineering Center, Massachusetts Institute of Technology, Room 56-469, Cambridge, Massachusetts 02139, USA.
Stephanopoulos
G
G
D000818:Animals; D016466:CHO Cells; D002453:Cell Cycle; D016923:Cell Death / Q000235:genetics; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media; D019254:Genes, bcl-2; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism*; D006801:Humans; D014162:Transfection
D003470:Culture Media; D005973:Glutamine; D005974:Glutamate-Ammonia Ligase