TY - JOUR T1 - Mimicking Muscle Stem Cell Quiescence in Culture: Methods for Synchronization in Reversible Arrest. JF - Methods Mol Biol Y1 - 2017 A1 - Arora, Reety A1 - Rumman, Mohammed A1 - Venugopal, Nisha A1 - Gala, Hardik A1 - Dhawan, Jyotsna KW - Actins KW - Adult Stem Cells KW - Animals KW - Biomarkers KW - Cell Culture Techniques KW - Cell Differentiation KW - Cell Line KW - Cell Proliferation KW - Fluorescent Antibody Technique KW - Humans KW - Mice KW - Microscopy, Fluorescence KW - Muscle, Skeletal KW - Myoblasts KW - Resting Phase, Cell Cycle KW - Satellite Cells, Skeletal Muscle KW - Stem Cells AB -

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.

VL - 1556 ER -