Adaptation of mammalian cells to non-ammoniagenic media.
Although glutamine is used as a major substrate for the growth of mammalian cells in culture, it suffers from some disadvantages. Glutamine is deaminated through storage or by cellular metabolism, leading to the formation of ammonia which can result in growth inhibition. Non-ammoniagenic alternatives to glutamine have been investigated in an attempt to develop strategies for obtaining improved cell yields for ammonia sensitive cell lines. Glutamate is a suitable substitute for glutamine in some culture systems. A period of adaptation to glutamate is required during which the activity of glutamine synthetase and the rate of transport of glutamate both increase. The cell yield increases when the ammonia accumulation is decreased following culture supplementation with glutamate rather than glutamine. However some cell lines fail to adapt to growth in glutamate and this may be due to a low efficiency transport system. The glutamine-based dipeptides, ala-gln and gly-gln can substitute for glutamine in cultures of antibody-secreting hybridomas. The accumulation of ammonia in these cultures is less and cell yields in dipeptide-based media may be improved compared to glutamine-based controls. In murine hybridomas, a higher concentration of gly-gln is required to obtain comparable cell growth to ala-gln or gln-based cultures. This is attributed to a requirement for dipeptide hydrolysis catalyzed by an enzyme with higher affinity for ala-gln than gly-gln.
Butler
M
M
Department of Microbiology, University of Manitoba, Winnipeg, Canada.
Christie
A
A
D000222:Adaptation, Physiological; D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002522:Chlorocebus aethiops; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods*; D004151:Dipeptides / Q000378:metabolism*; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism*; D005973:Glutamine / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000276:immunology; D007700:Kinetics; D008322:Mammals; D013997:Time Factors; D014709:Vero Cells
D000911:Antibodies, Monoclonal; D003470:Culture Media; D004151:Dipeptides; D000643:Ammonium Chloride; D005973:Glutamine; D005974:Glutamate-Ammonia Ligase