Metabolism of PER.C6™ cells cultivated under fed-batch conditions at low glucose and glutamine levels
Metabolism of PER.C6™ cells cultivated under fed-batch conditions at low glucose and glutamine levels
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DOI：
10.1002/bit.20890
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发表时间：
2006-05-05
影响因子：
3.8
通讯作者：
Goochee, CF
中科院分区：
文献类型：
--
作者：
Maranga, L;Goochee, CF
这是第一个研究检查PER. C6（TM）细胞葡萄糖/能量和谷氨酰胺代谢的补料分批培养物在控制的低谷氨酰胺，低葡萄糖，和同时低葡萄糖和低谷氨酰胺水平。在两升规模的无血清悬浮生物反应器中研究PER. C6（TM）细胞代谢。葡萄糖和/或谷氨酰胺浓度的控制对细胞代谢具有显著影响，导致营养物利用效率提高，副产物合成改变，同时对细胞生长速率没有影响。在0.25 mM的受控谷氨酰胺浓度下培养细胞使q（Gln）和q（NH 4+）降低约30%，q（Ala）降低85%，q（NEAA）降低50%。通过最小化谷氨酰胺的自发化学降解，谷氨酰胺的红色批次控制也将铵离子的总体积累降低了约50%。未观察到对葡萄糖/能量代谢的重大影响。在0.5mM的葡萄糖浓度下培养细胞使q（Glc）降低约50%并且消除乳酸积累。细胞表现出完全氧化代谢，Y-O2/Glc约为6 mol/mol。然而，尽管qGln没有增加，但也观察到铵离子积累和YNH 4 +/（Gln）增加。使用同时控制葡萄糖和谷氨酰胺的分批补料实现了PER. C6（TM）细胞对乳酸和4-铵离子积累的有效控制。获得了完全氧化的葡萄糖代谢和完全消除的乳酸产生。qGln值再次降低，尽管与分批培养相比q（NH 4+）增加，但铵离子水平通常低于分批培养中的相应水平，并且非必需氨基酸（NEAA）的积累减少约50%。总之，该研究表明，通过在葡萄糖和谷氨酰胺的毫摩尔受控水平下分批补料培养细胞，PER. C6（TM）细胞代谢可以被限制在具有提高的营养物利用效率的状态。此外，PER. C6（TM）细胞属于少数种类的哺乳动物细胞系，其中谷氨酰胺在能量代谢中起次要作用。(c)2006 Wiley Periodicals，Inc.
This is the first study to examine PER.C6((TM)) cell glucose/energy and glutamine metabolism with fed-batch cultures at controlled low glutamine, low glucose, and simultaneous low glucose and low glutamine levels. PER.C6 (TM) cell metabolism was investigated in serum-free suspension bioreactors at two-liter scale. Control of glucose and/or glutamine concentrations had a significant effect on cellular metabolism leading to an increased efficiency of nutrient utilization, altered byproduct synthesis, while having no effect on cell growth rate. Cultivating cells at a controlled glutamine concentration of 0.25 mM reduced q(Gln) and q(NH4+) by approximately 30%, q(Ala), 85%, and q(NEAA) 50%. The red-batch control of glutamine also reduced the overall accumulation of ammonium ion by approximately 50% by minimizing the spontaneous chemical degradation of glutamine. No major impact upon glucose/energy metabolism was observed. Cultivating cells at a glucose concentration of 0.5 mM reduced q(Glc) about 50% and eliminated lactate accumulation. Cells exhibited a fully oxidative metabolism with Y-O2/Glc of approximately 6 mol/mol. However, despite no increase in qGln, an increased ammonium ion accumulation and YNH4+/(Gln) were also observed. Effective control of lactate and 4 ammonium ion accumulation by PER.C6 (TM) cells was achieved using fed-batch with simultaneously controlled glucose and glutamine. A fully oxidative glucose metabolism and a complete elimination of lactate production were obtained. The qGln value was again reduced and, despite an increased q(NH4+) compared with batch culture, ammonium ion levels were typically lower than corresponding ones in batch cultures, and the accumulation of non-essential amino acids (NEAA) was reduced about 50%. In conclusion, this study shows that PER.C6 (TM) cell metabolism can be confined to a state with improved efficiencies of nutrient utilization by cultivating cells in fed-batch at millimolar controlled levels of glucose and glutamine. In addition, PER.C6 (TM) cells fall into a minority category of mammalian cell lines for which glutamine plays a minor role in energy metabolism. (c) 2006 Wiley Periodicals, Inc.