Linked Life Data

14654372

Source:http://linkedlifedata.com/resource/pubmed/id/14654372

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
12
pubmed:dateCreated
2003-12-5
pubmed:abstractText
Mouse embryonic stem (ES) cells easily differentiate towards the cardiac lineage making them suitable as an in vitro model to study cardiogenesis and as a potential source of transplantable cells. In this study, we show by in situ hybridisation that about 30% of the volume of cultures of differentiating ES cells consists of cardiomyocytes. RT-PCR analyses showed that the transcription factors Nkx2.5, Gata4, Mef2c and Irx4 were expressed at levels in the same order of magnitude as the levels observed in embryonic, neonatal and adult hearts. Atrial natriuretic factor and Connexin 40, associated with chamber formation in vivo, are expressed at relatively low levels, similar to those observed at early heart development in vivo. To facilitate the isolation of ES cell-derived cardiomyocytes, a cell line was constructed by stable transfection of the aminoglycoside phosphotransferase cDNA driven by the cardiac-specific distant upstream part of the Na(+)/Ca(2+) exchanger promoter. To accomplish single-copy integration, the construct was inserted into the hypoxanthine phosphoribosyltransferase locus of HM1 ES cells by homologous recombination. Cardiac-specific resistance to G418-sulphate (neomycin) allowed isolation of a pure population of cardiomyocytes. Genetically selected and unselected cell populations were characterised electrophysiologically using patch clamp. To explore whether clusters of cells have a similar differentiation profile, action potentials (APs) were measured in aggregates of differentiating ES cells, using a new method based on the voltage-dependent fluorescent dye di-4-ANEPPS. Both whole-cell recordings using patch-clamp and optical measurements with di-4-ANEPPS of the AP showed that upstroke velocity increases and AP duration decreases with differentiation time, accompanied by a decrease in AP interval, suggesting the initiation of the developmental programme underlying the formation of chamber myocardium.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0022-2828
pubmed:author
pubmed:issnType
Print
pubmed:volume
35
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1461-72
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:14654372-Action Potentials, pubmed-meshheading:14654372-Animals, pubmed-meshheading:14654372-Cell Differentiation, pubmed-meshheading:14654372-Cell Lineage, pubmed-meshheading:14654372-Cells, Cultured, pubmed-meshheading:14654372-Fluorescent Dyes, pubmed-meshheading:14654372-Immunohistochemistry, pubmed-meshheading:14654372-In Situ Hybridization, pubmed-meshheading:14654372-Mice, pubmed-meshheading:14654372-Mice, Inbred Strains, pubmed-meshheading:14654372-Myocardium, pubmed-meshheading:14654372-Myocytes, Cardiac, pubmed-meshheading:14654372-Patch-Clamp Techniques, pubmed-meshheading:14654372-Pyridinium Compounds, pubmed-meshheading:14654372-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:14654372-Selection, Genetic, pubmed-meshheading:14654372-Stem Cells, pubmed-meshheading:14654372-Transcription Factors, pubmed-meshheading:14654372-Transfection
pubmed:year
2003
pubmed:articleTitle
Cardiomyocytes purified from differentiated embryonic stem cells exhibit characteristics of early chamber myocardium.
pubmed:affiliation
Experimental and Molecular Cardiology Group, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't