Abstract
The maintenance of adult stem cells in their normal quiescent state depends on intrinsic factors and extrinsic signals originating from their microenvironment (also known as the stem cell niche). In skeletal muscle, its stem cells (satellite cells) lose their regenerative potential with aging, and this has been attributed, at least in part, to both age-associated changes in the satellite cells as in the niche cells, which include resident fibro-adipogenic progenitors (FAPs), macrophages, and endothelial cells, among others. To understand the regenerative decline of skeletal muscle with aging, there is a need for methods to specifically isolate stem and niche cells from resting muscle. Here we describe a fluorescence-activated cell sorting (FACS) protocol to simultaneously isolate discrete populations of satellite cells and niche cells from skeletal muscle of aging mice.
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Acknowledgments
Work in the authors’ laboratory has been supported by the Spanish Ministry of Science, Innovation and Universities, Spain (grant SAF2015-67369-R; and SAF 2015-70270-REDT, a María de Maeztu Unit of Excellence award to UPF [MDM-2014-0370], and a Severo Ochoa Center of Excellence award to the CNIC [SEV-2015-0505]), the UPF-CNIC collaboration agreement, ERC-2016-AdG-741966, La Caixa-HEALTH, AFM, MDA, and H2020-UPGRADE. V.M is recipient of a FPI predoctoral fellowship.
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Perdiguero, E., Moiseeva, V., Muñoz-Cánoves, P. (2019). Simultaneous Isolation of Stem and Niche Cells of Skeletal Muscle: Applicability for Aging Studies. In: Turksen, K. (eds) Stem Cells and Aging . Methods in Molecular Biology, vol 2045. Humana, New York, NY. https://doi.org/10.1007/7651_2019_210
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DOI: https://doi.org/10.1007/7651_2019_210
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