监测小鼠胚胎干细胞（mESCs）中的线粒体功能。

Monitoring Mitochondrial Function in Mouse Embryonic Stem Cells (mESCs).

机构信息

Department of Life Sciences, University of Coimbra, Coimbra, Portugal.

CNC-Center for Neuroscience and Cell Biology, CIBB, Azinhaga de Santa Comba, Polo 3, University of Coimbra, Coimbra, Portugal.

出版信息

Methods Mol Biol. 2021;2310:47-56. doi: 10.1007/978-1-0716-1433-4_4.

DOI:10.1007/978-1-0716-1433-4_4

PMID:34095997

Abstract

Mouse embryonic stem cells (mESCs) can be grown in culture, recapitulating the different states of pluripotency of their in vivo counterparts, with notably different metabolic profiles. mESCs in a naïve pluripotent state present an ambivalent metabolism, using both glycolysis and oxidative phosphorylation as energy sources. Here, we describe a method to evaluate the oxidative function of naïve mESCs using the Seahorse Extracellular Flux Analyzer coupled to flow cytometry analysis of mitochondrial transmembrane potential using the TMRM fluorescence probe, thus assessing both oxygen consumption and mitochondrial membrane potential. This may be a useful protocol for understanding how mitochondrial oxidative function and potential of mESCs change in certain circumstances, and how is it related with various pluripotency/differentiation phenotypes.

摘要

小鼠胚胎干细胞（mESCs）可以在培养中生长，重现其体内对应物的不同多能状态，具有明显不同的代谢特征。处于原始多能状态的 mESCs 表现出矛盾的代谢方式，同时使用糖酵解和氧化磷酸化作为能量来源。在这里，我们描述了一种使用 Seahorse 细胞外通量分析仪结合 TMRM 荧光探针流式细胞术分析线粒体膜电位来评估原始 mESCs 的氧化功能的方法，从而评估氧消耗和线粒体膜电位。这可能是一种有用的方法，可以帮助理解 mESCs 的线粒体氧化功能和潜力在某些情况下如何变化，以及它与各种多能性/分化表型有何关系。