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

• Published in: PloS one Vol. 7; no. 12; p. 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: Journal Article
• Language: English
• Published: United States Public Library of Science 19.12.2012; Public Library of Science (PLoS)
• 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; Australia
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0052214

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.