Abstract
Skeletal muscle harbors several types of cells, among which a population of progenitors committed to the adipogenic lineage has only recently been identified. Potential sources of white and brown adipocytes, the latter representing a potential target to treat obesity, are of considerable interest to the field. Fluorescence-activated cell sorting (FACS) provides an elegant strategy for prospective isolation of closely defined cell populations. Here we describe a flow cytometric method to isolate muscle-resident adipogenic progenitor cells with a default potential to undergo white adipogenesis. We further describe an approach to induce commitment to a lineage of brown-like adipocytes upon exposure to bone morphogenetic protein 7 (BMP7).
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References
Mauro A (1961) Satellite cell of skeletal muscle fibers. J Biophys Biochem Cytol 9:493–495
Hawke TJ, Garry DJ (2001) Myogenic satellite cells: physiology to molecular biology. J Appl Physiol 91:534–551
Zammit PS, Partridge TA, Yablonka-Reuveni Z (2006) The skeletal muscle satellite cell: the stem cell that came in from the cold. J Histochem Cytochem 54:1177–1191
Uezumi A, Fukada S, Yamamoto N et al (2010) Mesenchymal progenitors distinct from satellite cells contribute to ectopic fat cell formation in skeletal muscle. Nat Cell Biol 12:143–152
Joe AWB, Yi L, Natarajan A et al (2010) Muscle injury activates resident fibro/adipogenic progenitors that facilitate myogenesis. Nat Cell Biol 12:153-U144
Schulz TJ, Huang TL, Tran TT et al (2011) Identification of inducible brown adipocyte progenitors residing in skeletal muscle and white fat. Proc Natl Acad Sci U S A 108:143–148
Cerletti M, Jurga S, Witczak CA et al (2008) Highly efficient, functional engraftment of skeletal muscle stem cells in dystrophic muscles. Cell 134:37–47
Heredia JE, Mukundan L, Chen FM et al (2013) Type 2 innate signals stimulate fibro/adipogenic progenitors to facilitate muscle regeneration. Cell 153:376–388
Mozzetta C, Consalvi S, Saccone V et al (2013) Fibroadipogenic progenitors mediate the ability of HDAC inhibitors to promote regeneration in dystrophic muscles of young, but not old Mdx mice. EMBO Mol Med 5:626–639
Almind K, Manieri M, Sivitz WI et al (2007) Ectopic brown adipose tissue in muscle provides a mechanism for differences in risk of metabolic syndrome in mice. Proc Natl Acad Sci U S A 104:2366–2371
Qu Z, Balkir L, van Deutekom JC et al (1998) Development of approaches to improve cell survival in myoblast transfer therapy. J Cell Biol 142:1257–1267
Rando TA, Blau HM (1994) Primary mouse myoblast purification, characterization, and transplantation for cell-mediated gene therapy. J Cell Biol 125:1275–1287
Richler C, Yaffe D (1970) The in vitro cultivation and differentiation capacities of myogenic cell lines. Dev Biol 23:1–22
Arsic N, Mamaeva D, Lamb NJ et al (2008) Muscle-derived stem cells isolated as non-adherent population give rise to cardiac, skeletal muscle and neural lineages. Exp Cell Res 314:1266–1280
Qu-Petersen ZQ, Deasy B, Jankowski R et al (2002) Identification of a novel population of muscle stem cells in mice: potential for muscle regeneration. J Cell Biol 157:851–864
Gharaibeh B, Lu A, Tebbets J et al (2008) Isolation of a slowly adhering cell fraction containing stem cells from murine skeletal muscle by the preplate technique. Nat Protoc 3:1501–1509
Jankowski RJ, Haluszczak C, Trucco M et al (2001) Flow cytometric characterization of myogenic cell populations obtained via the preplate technique: potential for rapid isolation of muscle-derived stem cells. Hum Gene Ther 12:619–628
Conboy IM, Conboy MJ, Smythe GM et al (2003) Notch-mediated restoration of regenerative potential to aged muscle. Science 302:1575–1577
Sherwood RI, Christensen JL, Conboy IM et al (2004) Isolation of adult mouse myogenic progenitors: functional heterogeneity of cells within and engrafting skeletal muscle. Cell 119:543–554
Steenhuis P, Pettway GJ, Ignelzi MA Jr (2008) Cell surface expression of stem cell antigen-1 (Sca-1) distinguishes osteo-, chondro-, and adipoprogenitors in fetal mouse calvaria. Calcif Tissue Int 82:44–56
Pisani DF, Clement N, Loubat A et al (2010) Hierarchization of myogenic and adipogenic progenitors within human skeletal muscle. Stem Cells 28:2182–2194
Ieronimakis N, Balasundaram G, Reyes M (2008) Direct isolation, culture and transplant of mouse skeletal muscle derived endothelial cells with angiogenic potential. PLoS One 3. doi:10.1371/journal.pone.0001753
Pasut A, Oleynik P, Rudnicki MA (2012) Isolation of muscle stem cells by fluorescence activated cell sorting cytometry. Methods Mol Biol 798:53–64
Ratajczak MZ, Majka M, Kucia M et al (2003) Expression of functional CXCR4 by muscle satellite cells and secretion of SDF-1 by muscle-derived fibroblasts is associated with the presence of both muscle progenitors in bone marrow and hematopoietic stem/progenitor cells in muscles. Stem Cells 21:363–371
Beauchamp JR, Heslop L, Yu DS et al (2000) Expression of CD34 and Myf5 defines the majority of quiescent adult skeletal muscle satellite cells. J Cell Biol 151:1221–1234
Blanco-Bose WE, Yao CC, Kramer RH et al (2001) Purification of mouse primary myoblasts based on alpha 7 integrin expression. Exp Cell Res 265:212–220
Hulspas R (2010) Titration of fluorochrome-conjugated antibodies for labeling cell surface markers on live cells. Curr Protoc Cytom 54:6.29.1–6.29.9
Petrovic N, Walden TB, Shabalina IG et al (2010) Chronic peroxisome proliferator-activated receptor gamma (PPAR gamma) activation of epididymally derived white adipocyte cultures reveals a population of thermogenically competent, UCP1-containing adipocytes molecularly distinct from classic brown adipocytes. J Biol Chem 285:7153–7164
Bostrom P, Wu J, Jedrychowski MP et al (2012) A PGC1-alpha-dependent myokine that drives brown-fat-like development of white fat and thermogenesis. Nature 481:463–468
Fisher FM, Kleiner S, Douris N et al (2012) FGF21 regulates PGC-1alpha and browning of white adipose tissues in adaptive thermogenesis. Genes Dev 26:271–281
Rodeheffer MS, Birsoy K, Friedman JM (2008) Identification of white adipocyte progenitor cells in vivo. Cell 135:240–249
Acknowledgments
This work was supported by grants from the German Research Foundation (DFG; grant # SCHU 2445/2-1) and the European Research Council (grant # ERC-StG-2012-311082) to TJS.
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Lau, AM., Tseng, YH., Schulz, T.J. (2014). Adipogenic Fate Commitment of Muscle-Derived Progenitor Cells: Isolation, Culture, and Differentiation. In: Christ, B., Oerlecke, J., Stock, P. (eds) Animal Models for Stem Cell Therapy. Methods in Molecular Biology, vol 1213. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1453-1_19
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DOI: https://doi.org/10.1007/978-1-4939-1453-1_19
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