Analysis of cellular metabolism of hybridoma cells at distinct physiological states
Analysis of cellular metabolism of hybridoma cells at distinct physiological states
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DOI：
10.1263/jbb.95.317
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发表时间：
2003-04-01
影响因子：
2.8
通讯作者：
Hu, WS
中科院分区：
文献类型：
--
作者：
Gambhir, A;Korke, R;Hu, WS
杂交瘤细胞在连续培养中培养在化学定义的培养基中。这些培养达到了不同的稳定状态，其特点是不同的细胞代谢，这取决于导致稳定状态的培养条件。这些具有不同代谢的稳定状态的特征是乳酸产生与葡萄糖消耗的化学计量比(DeltaL/DeltaG)不同。当DeltaL/DeltaG降低时，葡萄糖、谷氨酰胺等氨基酸的比消耗率降低。这些稳定状态不具有几个离散值的DeltaL/DeltaG，相反，它们从高DeltaL/DeltaG状态(1.0)到中间状态(0.1小于或等于1.0)，并且在低DeltaL/DeltaG状态(0.1)时甚至进一步减小。进行代谢通量分析，以比较代表这三种不同代谢状态的细胞的能量代谢。通过使用化学定义的介质，促进了碳和氮的物质平衡。对生物量的形成进行了系统的估算。结果表明，所有糖酵解和TCA循环通量都随着DeltaL/DeltaG的减小而减小。在低DeltaL/DeltaG状态下，氨基酸比消耗率的降低伴随着丙酮酸周围所有通量的减少。分析还表明，TCA循环的外流形成丙酮酸，这有助于乳酸的形成，可能与高DeltaL/DeltaG状态下较高的氨基酸消耗率有关。综上所述，这些结果表明氨基酸代谢在减少哺乳动物细胞培养中乳酸的产生中起着重要作用。
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 (DeltaL/DeltaG). The specific consumption rates of glucose, glutamine and other amino acids are reduced when DeltaL/DeltaG reduces. Those steady states do not have a few discrete values of DeltaL/DeltaGs, rather they span from a high DeltaL/DeltaG state (> 1.0) to an intermediate state (0.1 less than or equal to DeltaL/DeltaG less than or equal to 1.0), and reduces even further at a low DeltaL/DeltaG 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 DeltaL/DeltaG decreases. At the low DeltaL/DeltaG 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 outflux 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 DeltaL/DeltaG state. Taken together the results suggest the amino acid metabolism plays an important role in reducing lactate production in mammalian cell culture.