Monitoring Mitochondrial Function in Mouse Embryonic Stem Cells (mESCs).
Metabolism
Mitochondrial Transmembrane potential
Pluripotency
Seahorse
Stem cells
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
7
6
2021
pubmed:
8
6
2021
medline:
11
8
2021
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 34095997
doi: 10.1007/978-1-0716-1433-4_4
doi:
Substances chimiques
Fluorescent Dyes
0
Rhodamines
0
tetramethylrhodamine methyl ester
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
47-56Références
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