title: Mimicking Muscle Stem Cell Quiescence in Culture: Methods for Synchronization in Reversible Arrest.
abstract: Growing evidence supports the view that in adult stem cells, the defining stem cell features of potency and self-renewal are associated with the quiescent state. Thus, uncovering the molecular logic of this reversibly arrested state underlies not only a fundamental understanding of adult tissue dynamics but also hopes for therapeutic regeneration and rejuvenation of damaged or aging tissue. A key question concerns how adult stem cells use quiescence to establish or reinforce the property of self-renewal. Since self-renewal is largely studied by assays that measure proliferation, the concept of self-renewal programs imposed during non-proliferating conditions is counterintuitive. However, there is increasing evidence generated by deconstructing the quiescent state that highlights how programs characteristic of this particular cell cycle exit may enhance stem cell capabilities, through both cell-intrinsic and extrinsic programs.Toward this end, culture models that recapitulate key aspects of stem cell quiescence are useful for molecular analysis to explore attributes and regulation of the quiescent state. In this chapter, we review the different methods used to generate homogeneous populations of quiescent muscle cells, largely by manipulating culture conditions that feed into core signaling programs that regulate the cell cycle. We also provide detailed protocols developed or refined in our lab over the past two decades.
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, Bangalore, India.|National Centre for Biological Sciences, Bangalore, India.
authors.forename: Reety
authors.initials: R
authors.lastname: Arora
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, Bangalore, India.|Manipal University, Manipal, India.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Rumman
authors.affiliation: CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India.
authors.forename: Nisha
authors.initials: N
authors.lastname: Venugopal
authors.affiliation: CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India.
authors.forename: Hardik
authors.initials: H
authors.lastname: Gala
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, Bangalore, India. jdhawan@ncbs.res.in.|CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India. jdhawan@ncbs.res.in.
authors.forename: Jyotsna
authors.initials: J
authors.lastname: Dhawan
mesh_terms: D000199:Actins / Q000378:metabolism; D053687:Adult Stem Cells / Q000166:cytology / Q000378:metabolism; D000818:Animals; D015415:Biomarkers; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D005455:Fluorescent Antibody Technique; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D018482:Muscle, Skeletal / Q000166:cytology*; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism; D016192:Resting Phase, Cell Cycle / Q000235:genetics; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D000199:Actins; D015415:Biomarkers
keywords: G0 phase; In vitro; Quiescence; Satellite cell; Stem cell