Bcl-xL expression interferes with the effects of L-glutamine supplementation on hybridoma cultures.
While feeding protocols and ectopic expression of anti-apoptotic genes have been used to improve the viability of hybridoma cell lines, the effect of the expression levels of survival genes on the behavior of hybridomas following nutrient supplementation is unknown. In this study, we compared the behavior of the Sp2/0-Ag14 hybridoma (Bcl-xL(low)) and the P3x63-Ag8.653 myeloma (Bcl-xL(high)) following culture supplementation with the amino acid L-glutamine (L-Gln). Our data revealed that L-Gln addition substantially increased Sp2/0-Ag14 cell viability and total cell density, concomitant with a decrease in the rate of cell death. This effect was not seen when other amino acids or D-glucose (D-Glc) replaced L-Gln. The improvement in the culture behavior of Sp2/0-Ag14 cells was attributed to a reduction in the rate of accumulation of apoptotic cells. On the other hand, L-Gln supplementation had only a limited effect on the growth of the P3x63-Ag8.653 cells. Interestingly, Sp2/0-Ag14 cells over-expressing Bcl-xL showed a culture behavior upon L-Gln complementation that was similar to the P3x63-Ag8.653 myeloma. These results suggest that the anti-apoptotic gene expression profile of hybridoma cells can markedly impact on the beneficial effects afforded by nutrient supplementation.
Charbonneau
Joel R
JR
Cellular Biochemistry Research Laboratory, Department of Chemistry and Biochemistry, Laurentian University, 935 Ramsey Lake Road, Sudbury, Ontario, Canada P3E 2C6.
Furtak
Teresa
T
Lefebvre
Jennie
J
Gauthier
Eric R
ER
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D005786:Gene Expression Regulation; D005973:Glutamine / Q000494:pharmacology*; D006825:Hybridomas / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D009101:Multiple Myeloma / Q000378:metabolism / Q000503:physiopathology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured / Q000378:metabolism; D051020:bcl-X Protein
C494812:Bcl2l1 protein, mouse; D003470:Culture Media; D019253:Proto-Oncogene Proteins c-bcl-2; D051020:bcl-X Protein; D005973:Glutamine