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首页> 外文期刊>Applied Microbiology and Biotechnology >Lactate and glucose concomitant consumption as a self-regulated pH detoxification mechanism in HEK293 cell cultures
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Lactate and glucose concomitant consumption as a self-regulated pH detoxification mechanism in HEK293 cell cultures

机译：乳酸和葡萄糖的伴随消耗作为HEK293细胞培养物中pH排毒的自我调节机制

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摘要

One of the most important limitations of mammalian cell-based processes is the secretion and accumulation of lactate as a by-product of their metabolism. Among the cell lines commonly used in industrial bioprocesses, HEK293 has been gaining importance over the last years. Up recently, HEK293 cells were known to consume lactate in late stages of cell culture usually when glucose and/or glutamine were depleted from media. Remarkably, in both scenarios, no significant cell growth was reported. However, we have observed a different metabolic behavior regarding lactate production and consumption in HEK293 cultures. HEK293 cells were able to co-metabolize glucose and lactate simultaneously, even in exponentially growing cell cultures. Our deep study of the effects of environmental conditions on lactate metabolism revealed that pH was the key to trigger the metabolic shift from lactate production to lactate and glucose concomitant consumption. Remarkably, this shift could be triggered at will when pH was set at 6.8. Even more interesting was the fact that lowering pH to 6.6 and supplementing media with exogenous lactate resulted in co-consumption of glucose and lactate from the beginning of cell culture, without affecting cell growth or protein productivity. On the contrary, cell growth was clearly hampered at this low pH if extracellular lactate was lacking. From our results, we hypothesize that HEK293 cells metabolize extracellular lactate as a strategy for pH detoxification, by means of co-transporting extracellular protons together with lactate into the cytosol. This novel hypothesis for unraveling lactate metabolism in HEK293 cells could open a door to re-direct genetic engineering strategies in order to obtain more efficient cell lines and also to further develop animal cell technology applications.
机译：基于哺乳动物细胞的过程的最重要限制之一是乳酸的分泌和积累，这是它们代谢的副产物。在工业生物过程中常用的细胞系中，HEK293在最近几年中变得越来越重要。最近，已知当培养基中的葡萄糖和/或谷氨酰胺耗尽时，HEK293细胞会在细胞培养的后期消耗乳酸。值得注意的是，在两种情况下，均未报告明显的细胞生长。但是，我们观察到关于HEK293培养物中乳酸的产生和消耗的不同代谢行为。即使在成倍增长的细胞培养物中，HEK293细胞也能够同时共代谢葡萄糖和乳酸。我们对环境条件对乳酸代谢的影响的深入研究表明，pH是触发代谢从乳酸生产向乳酸和葡萄糖伴随消耗的转变的关键。值得注意的是，当pH设置为6.8时，可以随意触发这种转变。更有趣的是，从细胞培养开始就将pH降低至6.6，并向培养基中添加外源乳酸会导致葡萄糖和乳酸的共同消耗，而不会影响细胞生长或蛋白质生产率。相反，如果缺乏细胞外乳酸，那么在这种低pH值下细胞生长明显受到阻碍。根据我们的结果，我们假设HEK293细胞通过将细胞外质子与乳酸一起共同转运到细胞质中来代谢细胞外乳酸作为pH排毒的策略。揭开HEK293细胞中乳酸代谢的这一新假说可为重新定向基因工程策略打开一扇门，以便获得更有效的细胞系并进一步发展动物细胞技术的应用。

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