Metabolic responses to different glucose and glutamine levels in baby hamster kidney cell culture

Abstract

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.

Erythropoietin production from CHO cells grown by continuous culture in a fluidized‐bed bioreactor

“…The accumulation of ammonia was minimized because of a low ammonia/glutamine coecient. Although the lactate/glucose coecient was relatively high compared to other mammalian cultures (Cruz et al, 1999;Yang and Butler, 2000;Zeng et al, 1998;Zhou et al, 1997), the accumulation of lactate is not so critical in a pH-controlIed environment (Hassell et al, 1991).…”

Growth characteristics of channel catfish ovary cells—influence of glucose and glutamine

“…It was reported that addition of glutamine inhibits glycolysis in the myeloma cell culture (Martinelle et al 1998) while in the cultures of BHK cells glutamine cannot be substituted by glucose but glucose can be substituted by glutamine (Cruz et al 1999). The consumption of glutamine and other amino acids brought about an accumulation of ammonium ions.…”

Metabolism of PER.C6TM cells cultivated under fed‐batch conditions at low glucose and glutamine levels

“…In BHK cells, the specific glutamine consumption rate seems to have a Michaelis-Menten dependence upon the residual glutamine concentration (Cruz et al, 1999a). A 50% decrease in glutamine consumption rate was observed in fed-batch cultures of HEK293 cells cultivated at millimolar levels of glutamine (Lee et al, 2003).…”