Analysis of CHO cells metabolic redistribution in a glutamate-based defined medium in continuous culture
Analysis of CHO cells metabolic redistribution in a glutamate-based defined medium in continuous culture
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DOI：
10.1021/bp0100981
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发表时间：
2001-11-01
影响因子：
2.9
通讯作者：
Gòdia, C
中科院分区：
文献类型：
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作者：
Altamirano, C;Illanes, A;Gòdia, C
在连续培养条件下，通过一系列葡萄糖下移和上移实验，研究了谷氨酸替代谷氨酰胺以及谷氨酸和葡萄糖代谢平衡对t-PA重组CHO细胞代谢再分配的影响。这些实验揭示了多个稳态的存在，实验数据用于进行代谢通量分析，以更好地了解细胞代谢及其再分布。葡萄糖进料速率的调节促进了葡萄糖和氮源利用的更高效率，同时降低了代谢副产物的产生，但这降低了t-PA比生产速率。在葡萄糖限制下的这种减少可以归因于细胞被迫有效地利用碳和能量源进行生长，从而削弱了代谢产物的产生。因此，在连续培养中，生长速率与碳源和能源可用性的结合决定了t-PA的生产水平。
The effect of glutamine replacement by glutamate and the balance between glutamate and glucose metabolism on the redistribution of t-PA-producing recombinant CHO cells metabolism is studied in a series of glucose shift down and shift up experiments in continuous culture. These experiments reveal the existence of multiple steady states, and experimental data are used to perform metabolic flux analysis to gain a better insight into cellular metabolism and its redistribution. Regulation of glucose feed rate promotes a higher efficiency of glucose and nitrogen source utilization, with lower production of metabolic byproducts, but this reduces t-PA specific production rate. This reduction under glucose limitation can be attributed to the fact that the cells are forced to efficiently utilize the carbon and energy source for growth, impairing the production of dispensable metabolites. It is, therefore, the combination of growth rate and carbon and energy source availability that determines the level of t-PA production in continuous culture.