Analysis of Cellular Metabolism of Hybridoma Cells at Distinct Physiological States

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Hybridoma cells were cultivated in a chemically defined medium in continuous cultures. These cultures reached different steady states marked by distinctive cell metabolism depending on the culture conditions leading to the steady state. Those steady states with different metabolism are characterized by different stoichiometric ratios of lactate production to glucose consumption (sΔL/ΔG). The specific consumption rates of glucose, glutamine and other amino acids are reduced when ΔL/ΔG reduces. Those steady states do not have a few discrete values of ΔL/ΔGs, rather they span from a high ΔL/ΔG state (>1.0) to an intermediate state (O.1 ≤ ΔL/ΔG ≤ 1.O), and reduces even further at a low ΔL/ΔG state (<0.1). Metabolic flux analysis was performed to compare energy metabolism of cells in cultures representing these three distinct metabolic states. The material balance on carbon and nitrogen was facilitated by the use of chemically defined medium. The formation of biomass was systematically estimated. It was revealed that all glycolysis and TCA cycle fluxes are reduced as ΔL/ΔG decreases. At the low ΔL/ΔG state, a reduction in amino acid specific consumption rate is accompanied by a reduction in all the fluxes around pyruvate. The analysis also shows that the out flux from the TCA cycle to form pyruvate, which contributes to lactate formation, is possibly linked to the higher consumption rate of amino acids at the high ΔL/ΔG state. Taken together the results suggest the amino acid metabolism plays an important role in reducing lactate production in mammalian cell culture.
社団法人日本生物工学会の論文
2003-04-25

Analysis of Cellular Metabolism of Hybridoma Cells at Distinct Physiological States

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