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首页> 外文期刊>The international journal of biochemistry and cell biology >Circumventing the Crabtree Effect: A method to induce lactate consumption and increase oxidative phosphorylation in cell culture
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Circumventing the Crabtree Effect: A method to induce lactate consumption and increase oxidative phosphorylation in cell culture

机译：规避Crabtree效应：一种诱导乳酸消耗并增加细胞培养物中氧化磷酸化的方法

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摘要

Most cells grown in glucose-containing medium generate almost all their ATP via glycolysis despite abundant oxygen supply and functional mitochondria, a phenomenon known as the Crabtree effect. By contrast, most cells within the body rely on mitochondrial oxidative phosphorylation (OXPHOS) to generate the bulk of their energy supply. Thus, when utilising the accessibility of cell culture to elucidate fundamental elements of mitochondria in health and disease, it is advantageous to adopt culture conditions under which the cells have greater reliance upon OXPHOS for the supply of their energy needs. Substituting galactose for glucose in the culture medium can provide these conditions, but additional benefit can be gained from alternate in vitro models. Herein we describe culture conditions in which complete autonomous depletion of medium glucose induces a lactate-consuming phase marked by increased MitoTracker Deep Red staining intensity, increased expression of Kreb's cycle proteins, increased expression of electron transport chain subunits, and increased sensitivity to the OXPHOS inhibitor rotenone. We propose these culture conditions represent an alternate accessible model for the in vitro study of cellular processes and diseases involving the mitochondrion without limitations incurred via the Crabtree effect. (C) 2016 The Author(s). Published by Elsevier Ltd.
机译：尽管有充足的氧气供应和功能性线粒体，在含葡萄糖的培养基中生长的大多数细胞几乎都通过糖酵解产生了所有的ATP，这种现象被称为Crabtree效应。相比之下，体内大多数细胞依靠线粒体的氧化磷酸化（OXPHOS）来产生大量的能量供应。因此，当利用细胞培养的可及性来阐明健康和疾病中线粒体的基本成分时，采用培养条件是有利的，在该条件下细胞更依赖于OXPHOS来满足其能量需求。用半乳糖替代培养基中的葡萄糖可以提供这些条件，但是可以从替代的体外模型中获得额外的好处。在本文中，我们描述了其中完全自主消耗中等葡萄糖会诱导乳酸消耗阶段的培养条件，该阶段的特征是MitoTracker深红色染色强度增加，Kreb循环蛋白表达增加，电子转运链亚基表达增加以及对OXPHOS抑制剂的敏感性增加鱼藤酮。我们建议这些培养条件代表一种替代的可访问模型，用于体外研究涉及线粒体的细胞过程和疾病，而不受Crabtree效应的限制。 （C）2016作者。由Elsevier Ltd.发布

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