2.50
Hdl Handle:
http://hdl.handle.net/10547/228316
Title:
The analysis of mitochondria and mitochondrial DNA in human embryonic stem cells.
Authors:
St John, Justin C.; Amaral, Alexandra; Bowles, Emma J.; Oliveira, João Facucho; Lloyd, Rhiannon E.I.; Freitas, Mariana; Gray, Heather L.; Navara, Christopher S.; Oliveira, Gisela; Schatten, Gerald P.; Spikings, Emma ( 0000-0001-8387-2098 ) ; Ramalho-Santos, João
Abstract:
As human embryonic stem cells (hESCs) undergo differentiation, they express genes characteristic of the lineage for which they are destined. However, fully differentiated individual cell types can be characterized by the number of mitochondria they possess and the copies of the mitochondrial genome per mitochondrion. These characteristics are indicative of a specific cell's requirement for adenosine triphosphate (ATP) and therefore cellular viability and function. Consequently, failure for an ESC to possess the full complement of mitochondria and mitochondrial DNA (mtDNA) could limit its final commitment to a particular fate. We describe a series of protocols that analyze the process of cellular mitochondrial and mtDNA differentiation during hESC differentiation. In addition, mtDNA transcription and replication are key events in cellular differentiation that require interaction between the nucleus and the mitochondrion. To this extent, we describe a series of protocols that analyze the initiation of these key events as hESCs progress from their undifferentiated state to the fully committed cell. Last, we describe real-time polymerase chain reaction protocols that allow both the identification of mtDNA copy number and determine whether mtDNA copy is uniform (homoplasmy) in its transmission or heterogeneous (heteroplasmy).
Affiliation:
Mitochondrial Reproductive Genetics Group, Division of Medical Sciences, The Medical School, University of Birmingham, UK.
Citation:
St John, J.C. et al (2006) 'The analysis of mitochondria and mitochondrial DNA in human embryonic stem cells' Methods in molecular biology 331:347-74
Publisher:
Humana Press
Journal:
Methods in molecular biology (Clifton, N.J.)
Issue Date:
2006
URI:
http://hdl.handle.net/10547/228316
DOI:
10.1385/1-59745-046-4:347
PubMed ID:
16881526
Additional Links:
http://www.ncbi.nlm.nih.gov/pubmed/16881526; http://link.springer.com/protocol/10.1385%2F1-59745-046-4%3A347
Type:
Book chapter
Language:
en
ISSN:
1064-3745
Appears in Collections:
Cell and Cryobiology Research Group

Full metadata record

DC FieldValue Language
dc.contributor.authorSt John, Justin C.en_GB
dc.contributor.authorAmaral, Alexandraen_GB
dc.contributor.authorBowles, Emma J.en_GB
dc.contributor.authorOliveira, João Facuchoen_GB
dc.contributor.authorLloyd, Rhiannon E.I.en_GB
dc.contributor.authorFreitas, Marianaen_GB
dc.contributor.authorGray, Heather L.en_GB
dc.contributor.authorNavara, Christopher S.en_GB
dc.contributor.authorOliveira, Giselaen_GB
dc.contributor.authorSchatten, Gerald P.en_GB
dc.contributor.authorSpikings, Emmaen_GB
dc.contributor.authorRamalho-Santos, Joãoen_GB
dc.date.accessioned2012-06-11T10:24:17Zen
dc.date.available2012-06-11T10:24:17Zen
dc.date.issued2006en
dc.identifier.citationSt John, J.C. et al (2006) 'The analysis of mitochondria and mitochondrial DNA in human embryonic stem cells' Methods in molecular biology 331:347-74en_GB
dc.identifier.issn1064-3745en
dc.identifier.pmid16881526en
dc.identifier.doi10.1385/1-59745-046-4:347en
dc.identifier.urihttp://hdl.handle.net/10547/228316en
dc.description.abstractAs human embryonic stem cells (hESCs) undergo differentiation, they express genes characteristic of the lineage for which they are destined. However, fully differentiated individual cell types can be characterized by the number of mitochondria they possess and the copies of the mitochondrial genome per mitochondrion. These characteristics are indicative of a specific cell's requirement for adenosine triphosphate (ATP) and therefore cellular viability and function. Consequently, failure for an ESC to possess the full complement of mitochondria and mitochondrial DNA (mtDNA) could limit its final commitment to a particular fate. We describe a series of protocols that analyze the process of cellular mitochondrial and mtDNA differentiation during hESC differentiation. In addition, mtDNA transcription and replication are key events in cellular differentiation that require interaction between the nucleus and the mitochondrion. To this extent, we describe a series of protocols that analyze the initiation of these key events as hESCs progress from their undifferentiated state to the fully committed cell. Last, we describe real-time polymerase chain reaction protocols that allow both the identification of mtDNA copy number and determine whether mtDNA copy is uniform (homoplasmy) in its transmission or heterogeneous (heteroplasmy).en_GB
dc.language.isoenen
dc.publisherHumana Pressen_GB
dc.relation.urlhttp://www.ncbi.nlm.nih.gov/pubmed/16881526en_GB
dc.relation.urlhttp://link.springer.com/protocol/10.1385%2F1-59745-046-4%3A347en
dc.subjectMitochondriaen_GB
dc.subjectmitochondrial DNAen_GB
dc.subjecthuman embryonic stem cellsen_GB
dc.subjectdifferentiationen_GB
dc.subjectcardiomyocytesen_GB
dc.subjecthomoplasmyen_GB
dc.subjectheteroplasmyen_GB
dc.subjecttranscription and replicationen_GB
dc.subject.meshBenzimidazolesen
dc.subject.meshCarbocyaninesen
dc.subject.meshCell Culture Techniquesen
dc.subject.meshCell Differentiationen
dc.subject.meshCell Divisionen
dc.subject.meshDNA, Mitochondrialen
dc.subject.meshFluorescent Dyesen
dc.subject.meshGene Dosageen
dc.subject.meshGenome, Humanen
dc.subject.meshHumansen
dc.subject.meshImmunohistochemistryen
dc.subject.meshMembrane Potentialsen
dc.subject.meshMicroscopy, Fluorescenceen
dc.subject.meshMitochondriaen
dc.subject.meshMyocytes, Cardiacen
dc.subject.meshPluripotent Stem Cellsen
dc.subject.meshRNAen
dc.subject.meshRNA, Small Interferingen
dc.subject.meshReverse Transcriptase Polymerase Chain Reactionen
dc.subject.meshSequence Analysis, DNAen
dc.subject.meshTranscription, Geneticen
dc.subject.meshTransfectionen
dc.titleThe analysis of mitochondria and mitochondrial DNA in human embryonic stem cells.en
dc.typeBook chapteren
dc.contributor.departmentMitochondrial Reproductive Genetics Group, Division of Medical Sciences, The Medical School, University of Birmingham, UK.en_GB
dc.identifier.journalMethods in molecular biology (Clifton, N.J.)en_GB

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