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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Published in | PloS one Vol. 7; no. 12; p. e52214 |
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Main Authors | , , , , , , , |
Format | Journal Article |
Language | English |
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Public Library of Science
19.12.2012
Public Library of Science (PLoS) |
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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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AbstractList | 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. |
Audience | Academic |
Author | Gray, Peter P Prowse, Andrew B J Elliott, David A Chong, Fenny Elefanty, Andrew G Stanley, Edouard G Munro, Trent P Osborne, Geoffrey W |
AuthorAffiliation | 4 Murdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, Australia University of Cincinnati, United States of America 2 Monash Immunology and Stem Cell Laboratories, Monash University, Clayton, Australia 3 Queensland Brain Institute, The University of Queensland, St. Lucia, Australia 1 The Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, Australia |
AuthorAffiliation_xml | – name: 3 Queensland Brain Institute, The University of Queensland, St. Lucia, Australia – name: 1 The Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, Australia – name: 2 Monash Immunology and Stem Cell Laboratories, Monash University, Clayton, Australia – name: 4 Murdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, Australia – name: University of Cincinnati, United States of America |
Author_xml | – sequence: 1 givenname: Andrew B J surname: Prowse fullname: Prowse, Andrew B J email: a.prowse@uq.edu.au organization: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, Australia. a.prowse@uq.edu.au – sequence: 2 givenname: Fenny surname: Chong fullname: Chong, Fenny – sequence: 3 givenname: David A surname: Elliott fullname: Elliott, David A – sequence: 4 givenname: Andrew G surname: Elefanty fullname: Elefanty, Andrew G – sequence: 5 givenname: Edouard G surname: Stanley fullname: Stanley, Edouard G – sequence: 6 givenname: Peter P surname: Gray fullname: Gray, Peter P – sequence: 7 givenname: Trent P surname: Munro fullname: Munro, Trent P – sequence: 8 givenname: Geoffrey W surname: Osborne fullname: Osborne, Geoffrey W |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/23284940$$D View this record in MEDLINE/PubMed |
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Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Conceived and designed the experiments: AP FC TM DE AE ES GO. Performed the experiments: AP FC GO. Analyzed the data: AP FC GO DE AE ES. Contributed reagents/materials/analysis tools: AP AE ES PG GO. Wrote the paper: AP DE GO TM. Competing Interests: The authors have declared that no competing interests exist. |
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SubjectTerms | 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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Title | Analysis of mitochondrial function and localisation during human embryonic stem cell differentiation in vitro |
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