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Analysis of Mitochondrial Function and Localisation during Human Embryonic Stem Cell Differentiation In Vitro

Authors
Journal
PLoS ONE
1932-6203
Publisher
Public Library of Science
Publication Date
Volume
7
Issue
12
Identifiers
DOI: 10.1371/journal.pone.0052214
Keywords
  • Research Article
  • Biology
  • Biotechnology
  • Small Molecules
  • Developmental Biology
  • Stem Cells
  • Cell Potency
  • Stem Cell Lines
  • Cell Differentiation
  • Molecular Cell Biology
  • Cellular Structures
  • Subcellular Organelles
  • Cellular Types
  • Embryonic Stem Cells
Disciplines
  • Biology
  • Medicine

Abstract

Human embryonic stem cell (hESC) derivatives show promise as viable cell therapy options for multiple disorders in different tissues. Recent advances in stem cell biology have lead to the reliable production and detailed molecular characterisation of a range of cell-types. However, the role of mitochondria during differentiation has yet to be fully elucidated. Mitochondria mediate a cells response to altered energy requirements (e.g. cardiomyocyte contraction) and, as such, the mitochondrial phenotype is likely to change during the dynamic process of hESC differentiation. We demonstrate that manipulating mitochondrial biogenesis alters mesendoderm commitment. To investigate mitochondrial localisation during early lineage specification of hESCs we developed a mitochondrial reporter line, KMEL2, in which sequences encoding the green fluorescent protein (GFP) are targeted to the mitochondria. Differentiation of KMEL2 lines into the three germ layers showed that the mitochondria in these differentiated progeny are GFP positive. Therefore, KMEL2 hESCs facilitate the study of mitochondria in a range of cell types and, importantly, permit real-time analysis of mitochondria via the GFP tag.

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