Monitoring hybridoma metabolism in continuous suspension culture at the intracellular level. I. Steady-state responses to different glutamine feed concentrations.
A model mouse hybridoma cell line was grown in continuous culture experiments in a serum-free low-protein lipid-free medium. The steady-state responses of cell numbers, extra- and intracellular metabolite concentrations, substrate and (by) product consumption/production rates, and yield coefficients were investigated as a function of step changes in the glutamine concentration of the feed medium. In addition to the commonly performed analysis of metabolites in culture supernatants, we prepared perchloric acid extracts of cells and determined the amount and the composition of intracellular amino acids and organic acids. Significant differences were found with respect to intracellular metabolite pools for cells growing at nearly identical specific growth rates. To our knowledge this is the first time that data on the intracellular concentrations (pools) of amino acids and Krebs cycle intermediates are reported in the literature that were obtained under carefully defined culture conditions such as those attained in continuous culture experiments.
F. Hoffmann-La Roche Ltd., Pharmaceutical Research--New Technologies, Department PRTM, Basel, Switzerland.
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Keller
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002264:Carboxylic Acids / Q000378:metabolism; D002457:Cell Extracts / Q000737:chemistry; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free; D005973:Glutamine / Q000008:administration & dosage*; D006825:Hybridomas / Q000378:metabolism*; D051379:Mice; D008991:Monitoring, Physiologic; D013997:Time Factors
D000596:Amino Acids; D002264:Carboxylic Acids; D002457:Cell Extracts; D016895:Culture Media, Serum-Free; D005973:Glutamine