Extracellular Flux Analysis of Mitochondrial Function in Pluripotent Stem Cells

Enkhtuul Tsogtbaatar, Katherine Minter-Dykhouse, Alicia Saarinen, Clifford D.L. Folmes

Research output: Chapter in Book/Report/Conference proceedingChapter


Mitochondrial function and energy metabolism are increasingly recognized not only as regulators of pluripotent stem cell function and fate, but also as critical targets in disease pathogenesis and aging. Therefore across the downstream applications of pluripotent stem cells, including development and disease modeling, drug screening, and cell-based therapies, it is crucial to be able to measure mitochondrial function and metabolism in a high-throughput, real-time and label-free manner. Here we describe the application of Seahorse extracellular flux analysis to measure mitochondrial function in pluripotent stem cells and their derivatives. Specifically, we highlight two assays, the Mitochondrial Stress Test, which quantifies overall mitochondrial function including basal, maximal and ATP-couple oxygen consumption rates, and the Electron Transport Chain Complex Specific assay, that quantifies function of individual complexes within the electron transport chain.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages18
StatePublished - 2022

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029


  • Differentiation
  • Embryonic stem cells
  • Induced pluripotent stem cells
  • Mitochondrial respiration
  • Oxidative metabolism
  • Oxidative phosphorylation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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