In vitro induction of quiescence in isolated primary human myoblasts

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dc.contributor.authorGudagudi, Kirankumar B.en_ZA
dc.contributor.authord'Entrèves, Niccolò Passerinen_ZA
dc.contributor.authorWoudberg, Nicholas J.en_ZA
dc.contributor.authorSteyn, Paul J.en_ZA
dc.contributor.authorMyburgh, Kathryn H.en_ZA
dc.date.accessioned2022-06-23T09:09:45Z
dc.date.available2022-06-23T09:09:45Z
dc.date.issued2020-01-28
dc.descriptionCITATION: Gudagudi, K. B. et al. 2020. In vitro induction of quiescence in isolated primary human myoblasts. Cytotechnology, 72:189–202. doi:10.1007/s10616-019-00365-8
dc.descriptionThe original publication is available at https://www.springer.com/journal/10616/
dc.description.abstractAdult skeletal muscle stem cells, satellite cells, remain in an inactive or quiescent state in vivo under physiological conditions. Progression through the cell cycle, including activation of quiescent cells, is a tightly regulated process. Studies employing in vitro culture of satellite cells, primary human myoblasts (PHMs), necessitate isolation myoblasts from muscle biopsies. Further studies utilizing these cells should endeavour to represent their native in vivo characteristics as closely as possible, also considering variability between individual donors. This study demonstrates the approach of utilizing KnockOut™ Serum Replacement (KOSR)-supplemented culture media as a quiescence-induction media for 10 days in PHMs isolated and expanded from three different donors. Cell cycle analysis demonstrated that treatment resulted in an increase in G1 phase and decreased S phase proportions in all donors (p < 0.005). The proportions of cells in G1 and G2 phases differed in proliferating myoblasts when comparing donors (p < 0.05 to p < 0.005), but following KOSR treatment, the proportion of cells in G1 (p = 0.558), S (p = 0.606) and G2 phases (p = 0.884) were equivalent between donors. When cultured in the quiescence-induction media, expression of CD34 and Myf5 remained constant above > 98% over time from day 0 to day 10. In contrast activation (CD56), proliferation (Ki67) and myogenic marker MyoD decreased, indicated de-differentiation. Induction of quiescence was accompanied in all three clones by fold change in p21 mRNA greater than 3.5 and up to tenfold. After induction of quiescence, differentiation into myotubes was not affected. In conclusion, we describe a method to induce quiescence in PHMs from different donors.en_ZA
dc.description.urihttps://link.springer.com/article/10.1007/s10616-019-00365-8
dc.description.versionPublishers version
dc.format.extent14 pages : illustrationsen_ZA
dc.identifier.citationGudagudi, K. B. et al. 2020. In vitro induction of quiescence in isolated primary human myoblasts. Cytotechnology, 72:189–202. doi:10.1007/s10616-019-00365-8
dc.identifier.issn1573-0778 (online)
dc.identifier.issn0920-9069 (print)
dc.identifier.otherdoi:10.1007/s10616-019-00365-8
dc.identifier.urihttp://hdl.handle.net/10019.1/125399
dc.language.isoen_ZAen_ZA
dc.publisherSpringer Nature
dc.rights.holderAuthors retain copyright
dc.subjectCell cycle inhibitionen_ZA
dc.subjectIsolationen_ZA
dc.subjectPrimary human myoblastsen_ZA
dc.subjectQuiescenceen_ZA
dc.subjectMuscle cells -- Growth -- Regulationen_ZA
dc.subjectCell cycle -- Inhibitorsen_ZA
dc.titleIn vitro induction of quiescence in isolated primary human myoblastsen_ZA
dc.typeArticleen_ZA
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