Analysis of mitochondrial function and localisation during human embryonic stem cell differentiation in vitro

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 mitoc...

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Bibliographic Details
Published in:PloS one 2012-12, Vol.7 (12), p.e52214-e52214
Main Authors: Prowse, Andrew B J, Chong, Fenny, Elliott, David A, Elefanty, Andrew G, Stanley, Edouard G, Gray, Peter P, Munro, Trent P, Osborne, Geoffrey W
Format: Article
Language:English
Subjects:
Ataxia
Bioengineering
Biology
Biosynthesis
Cardiomyocytes
Cell differentiation
Cell Differentiation - physiology
Cell Line
Contraction
Deoxyribonucleic acid
Differentiation (biology)
DNA
Embryonic stem cells
Embryonic Stem Cells - cytology
Embryonic Stem Cells - metabolism
Energy requirements
Flow Cytometry
Fluorescence
Fluorescent Antibody Technique
Gene expression
Genomes
Genotype & phenotype
Green fluorescent protein
Humans
Immunology
Karyotype
Kinases
Laboratories
Localization
Mesendoderm
Metabolism
Mitochondria
Mitochondria - metabolism
Mitochondrial DNA
Nanotechnology
Oxidative Phosphorylation
Phosphorylation
Progeny
Proteins
Stem cell transplantation
Stem cells
Tissues
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0052214