Glutamine or glucose starvation in hybridoma cultures induces death receptor and mitochondrial apoptotic pathways.
Glutamine and glucose are often controlled at low levels in fed-batch strategies to limit ammonia and lactate accumulation and improve productivity of mammalian cell cultures. However, this risks triggering apoptosis if cells are depleted of glutamine or glucose. To examine the apoptosis cascade during glutamine or glucose limitation, the transcriptional profile of FAS, FASL, FADD, FLIP, BAX, p53 and PEG3 in CRL 1606 hybridoma culture was investigated using quantitative real-time PCR. Activities of caspases 2, 3, 8 and 9 were also analyzed. Increase in the activities of the caspases was observed with up-regulation in the expression of FAS (6-8-fold) and PEG3 (2.5-fold), suggesting that the cells experienced apoptotic cell death via both the death receptor and mitochondrial pathways.
Yeo
Jessna H M
JH
Bioprocessing Technology Institute, Agency for Science Technology and Research, A*STAR, 20 Biopolis Way #06-01, Centros, Singapore 138668.
Lo
Jennifer C Y
JC
Nissom
Peter M
PM
Wong
Victor V T
VV
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D019149:Bioreactors; D020169:Caspases / Q000378:metabolism; D018929:Cell Culture Techniques; D002460:Cell Line; D005947:Glucose / Q000172:deficiency*; D005973:Glutamine / Q000172:deficiency*; D006825:Hybridomas / Q000187:drug effects* / Q000378:metabolism; D051741:Kruppel-Like Transcription Factors; D051379:Mice; D008928:Mitochondria / Q000378:metabolism; D011494:Protein Kinases / Q000096:biosynthesis; D053218:Receptors, Death Domain / Q000096:biosynthesis*; D014157:Transcription Factors / Q000096:biosynthesis; D019014:fas Receptor / Q000096:biosynthesis
D051741:Kruppel-Like Transcription Factors; C097901:Peg3 protein, mouse; D053218:Receptors, Death Domain; D014157:Transcription Factors; D019014:fas Receptor; D005973:Glutamine; D011494:Protein Kinases; D020169:Caspases; D005947:Glucose