Single-fiber isolation and maintenance of satellite cell quiescence

dc.contributor.authorWozniak, AC
dc.contributor.authorAnderson, JE
dc.date.accessioned2007-10-09T18:47:37Z
dc.date.available2007-10-09T18:47:37Z
dc.date.issued2005-10-31
dc.description.abstractThe activity of satellite cells during myogenesis, development, or skeletal muscle regeneration is strongly modelled using cultures of single muscle fibers. However, there are variations in reported features of gene or protein expression as examined with single-fiber cultures. Here, we examined the potential differences in activation of satellite cells on normal mouse muscle fibers produced during a standard isolation protocol, with or without agitation during collagenase digestion. Activation was detected in satellite cells on fibers after 24 and 48 h of culture in basal growth medium using immunodetection of the incorporation of bromodeoxyuridine (BrdU) into DNA and quantification of the number of BrdU-positive cells per fiber. After 24 and 48 h in culture under nonactivating conditions, the number of activated (BrdU+) satellite cells was greater on fibers that had received gentle agitation during collagenase digestion than on those that were subject to digestion without agitation during isolation. The findings are interpreted to mean that at least some of the variation among published reports may derive from the application of various methods of fiber isolation. The information should be useful for maintaining satellite cell quiescence during studies of the regulatory steps that lead to satellite cell activation.en
dc.format.extent80278 bytes
dc.format.mimetypeapplication/pdf
dc.identifier.citation0829-8211; BIOCHEM CELL BIOL, OCT 2005, vol. 83, no. 5, p.674 to 676.en
dc.identifier.doihttp://dx.doi.org/10.1139/o05-046
dc.identifier.urihttp://hdl.handle.net/1993/2916
dc.language.isoengen_US
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dc.rightsopen accessen_US
dc.statusPeer revieweden
dc.subjectactivationen
dc.subjectskeletal muscleen
dc.subjectproliferationen
dc.subjectsingle-fiber cultureen
dc.subjectmyogenesisen
dc.subjectSKELETAL-MUSCLEen
dc.subjectIN-VIVOen
dc.subjectACTIVATIONen
dc.subjectEXPRESSIONen
dc.subjectCULTUREen
dc.subjectMYF5en
dc.titleSingle-fiber isolation and maintenance of satellite cell quiescenceen
dc.typejournal articleen_US
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