Decoupling cell growth and product formation in Chinese hamster ovary cells through metabolic control
Decoupling cell growth and product formation in Chinese hamster ovary cells through metabolic control
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DOI：
10.1002/bit.10096
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发表时间：
2001-11-01
影响因子：
3.8
通讯作者：
Gòdia, F
中科院分区：
文献类型：
--
作者：
Altamirano, C;Cairó, JJ;Gòdia, F
研究了中国仓鼠卵巢（CHO）细胞产生组织型纤溶酶原激活剂（t-PA）的培养策略。这种策略是基于用另一种代谢缓慢的化合物（特别是半乳糖）替代主要碳源葡萄糖。这种变化的引入允许细胞行为在各个水平上发生急剧变化。细胞生长在这种营养转移后停止，并且与仅基于葡萄糖利用的培养策略相比，细胞可以以低生长速率和高活力保持在长时间培养中。此外，细胞产生重组蛋白（本研究中的t-PA）的能力可以在整个半乳糖喂养期间保持。从代谢的角度来看，使用缓慢代谢的碳源（半乳糖）会导致乳酸盐、氨和一些氨基酸的产生发生重要变化。这种代谢转变的使用使得能够产生双相过程，其中第一阶段是葡萄糖上的细胞生长，第二阶段是半乳糖上的稳定期，这特别适合于灌注系统。(C)John Wiley & Sons，Inc.
The development of a strategy for the culture of Chinese hamster ovary (CHO) cells producing tissue plasminogen activator (t-PA) is investigated. This strategy is based on the replacement of the main carbon source, glucose, by another compound that is slowly metabolizable, particularly galactose. The introduction of this change allows for acute change in cell behavior at various levels. Cell growth is stopped after this nutrient shift, and the cells can be kept in long-duration culture at a low growth rate and high viability as compared with a culture strategy based solely on glucose utilization. Moreover, the capability of cells to produce recombinant proteins (t-PA in this work) can be maintained over the entire period of galactose feeding. From the metabolic point of view, use of a slowly metabolizable carbon source (galactose) introduces important changes in the production of lactate, ammonia, and some amino acids. The use of this metabolic shift enables the generation of biphasic processes, with a first phase with cell growth on glucose and a second stationary phase on galactose, which is particularly suited to perfusion systems. (C) 2001 John Wiley & Sons, Inc.