{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,13]],"date-time":"2026-11-13T02:32:36Z","timestamp":1794537156192,"version":"4.9.1"},"reference-count":44,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2013,1,24]],"date-time":"2013-01-24T00:00:00Z","timestamp":1358985600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J cachexia sarcopenia muscle"],"published-print":{"date-parts":[[2013,6]]},"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>During aging, there is a decreased ability to maintain skeletal muscle mass and function (sarcopenia). Such changes in skeletal muscle are also co\u2010morbidities of diseases including cancer, congestive heart failure and chronic obstructive pulmonary disease. The loss of muscle mass results in decreased strength and exercise tolerance and reduced ability to perform daily activities. Pharmacological agents addressing these pathologies could have significant clinical impact, but their identification requires understanding of mechanisms driving myotube formation (myogenesis) and atrophy and provision of relevant assays. The aim of this study was to develop robust in vitro methods to study human myogenesis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Satellite cells were isolated by digestion of post\u2010mortem skeletal muscle and selection using anti\u2010CD56 MicroBeads. CD56<jats:sup>+<\/jats:sup> cell\u2010derived myotubes were quantified by high content imaging of myosin heavy chains. TaqMan\u2010polymerase chain reaction arrays were used to quantify expression of 41 selected genes during differentiation. The effects of activin receptor agonists and tumour necrosis factor alpha (TNF\u03b1) on myogenesis and gene expression were characterised.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Large\u2010scale isolation of CD56<jats:sup>+<\/jats:sup> cells enabled development of a quantitative myogenesis assay with maximal myotube formation 3\u00a0days after initiating differentiation. Gene expression analysis demonstrated expression of 19 genes changed substantially during myogenesis. TNF\u03b1 and activin receptor agonists inhibited myogenesis and downregulated gene expression of muscle transcription factors, structural components and markers of oxidative phenotype, but only TNF\u03b1 increased expression of pro\u2010inflammatory markers.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>We have developed methods for large\u2010scale isolation of satellite cells from muscle and quantitative assays for studying human myogenesis. These systems may prove useful as part of a screening cascade designed to identify therapeutic agents for improving muscle function.<\/jats:p><\/jats:sec><jats:sec><jats:title>Electronic supplementary material<\/jats:title><jats:p>The online version of this article (doi:10.1007\/s13539\u2010012\u20100097\u2010z) contains supplementary material, which is available to authorized users.<\/jats:p><\/jats:sec>","DOI":"10.1007\/s13539-012-0097-z","type":"journal-article","created":{"date-parts":[[2013,1,23]],"date-time":"2013-01-23T16:38:09Z","timestamp":1358959089000},"page":"157-169","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":31,"title":["Large\u2010scale isolation of human skeletal muscle satellite cells from post\u2010mortem tissue and development of quantitative assays to evaluate modulators of myogenesis"],"prefix":"10.1002","volume":"4","author":[{"given":"Ian C.","family":"Scott","sequence":"first","affiliation":[{"name":"Respiratory and Inflammation, Bioscience Department, AstraZeneca R&amp;D Charnwood  Loughborough"},{"name":"Department of Respiratory Inflammation and Autoimmunity MedImmune  Milstein Building, Granta Park CB21 6GH Cambridge"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wendy","family":"Tomlinson","sequence":"additional","affiliation":[{"name":"Respiratory and Inflammation, Bioscience Department, AstraZeneca R&amp;D Charnwood  Loughborough"},{"name":"Mars Petcare  Waltham-on-the-Wolds LE14 4RT Melton Mowbray"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew","family":"Walding","sequence":"additional","affiliation":[{"name":"Respiratory and Inflammation, Bioscience Department, AstraZeneca R&amp;D Charnwood  Loughborough"},{"name":"Global Medicines Development, Global Clinical Development AstraZeneca  Alderley Park SK10 4TG 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