Metabolic responses to different glucose and glutamine levels in baby hamster kidney cell culture.
In this work, a BHK21 clone producing a recombinant antibody/cytokine fusion protein was used to study the dependence of cell metabolism on the glucose and glutamine levels in the culture medium. Results obtained indicate that both glucose and glutamine consumptions show a Michaelis-Menten dependence on glucose and glutamine concentrations respectively. A similar dependence is also observed for lactate and ammonia productions. The estimated value of the Michaelis constant for the dependence of lactate production on glucose (KLacGlc) was 1.4 +/- 0.1 mM and for the dependence of ammonia production on glutamine (KAmmGln) was 0.25 +/- 0.11 mM and 0.10 +/- 0.03 mM, at glucose concentrations of 0.28 mM and 5.6 mM respectively. At very low glucose concentrations, the glucose to lactate yield decreased markedly, showing a metabolic shift towards lower lactate production. This metabolic shift was also confirmed by the significant increase in the specific oxygen consumption rate also observed at low glucose concentrations. Although it was highly dependent on glucose concentration, the oxygen consumption also increased with the increase in glutamine concentration. At very low glutamine concentrations, the glutamine to ammonia yield increased, showing a more efficient glutamine metabolism.
Cruz
H J
HJ
IBET/ITQB-Instituto de Biologia Experimental e Tecnológica/Instituto de Tecnologia Química e Biológica, Oeiras, Portugal.
Ferreira
A S
AS
Freitas
C M
CM
Moreira
J L
JL
Carrondo
M J
MJ
D000641:Ammonia / Q000378:metabolism; D000818:Animals; D002460:Cell Line / Q000378:metabolism; D006224:Cricetinae; D003470:Culture Media / Q000737:chemistry; D005947:Glucose / Q000378:metabolism* / Q000494:pharmacology; D005973:Glutamine / Q000378:metabolism* / Q000494:pharmacology; D010101:Oxygen Consumption / Q000187:drug effects
D003470:Culture Media; D005973:Glutamine; D000641:Ammonia; D005947:Glucose