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Isolation and Culture of Quiescent Skeletal Muscle Satellite Cells

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Book cover Stem Cells and Tissue Repair

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2155))

Abstract

It has been shown that freshly isolated satellite cells from adult muscle constitute a stem cell-like population that exhibits more efficient engraftment and self-renewal activity in regenerating muscle than myoblast. Thus, purification of pure populations of quiescent satellite cells from adult skeletal muscle is highly necessary, not only for understanding the biology of satellite cells and myoblasts but also for improving cell-based therapies for muscle regeneration. This chapter describes a basic protocol used in our laboratory to isolate quiescent muscle satellite cells from adult skeletal muscle by enzymatic dissociation followed by a sequential magnetic-activated cell sorting (MACS). This method is cheap and fast providing and alternative procedure to other purification methods that require fluorescence-activated cell sorting (FACS) machines. Freshly isolated quiescent satellite cells purified by this method can be used in a broad range of experiments including cell transplantation for satellite cell self-renewal experiments or cell therapies.

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References

  1. Janssen I, Heymsfield SB, Wang ZM et al (2000) Skeletal muscle mass and distribution in 468 men and women aged 18-88 yr. J Appl Physiol 89:81–88

    Article  CAS  Google Scholar 

  2. Almada AE, Wagers AJ (2016) Molecular circuitry of stem cell fate in skeletal muscle regeneration, ageing and disease. Nat Rev Mol Cell Biol 17:267–279

    Article  CAS  Google Scholar 

  3. Vallejo D, Hernández-Torres F, Lozano-Velasco E et al (2018) PITX2 enhances the regenerative potential of dystrophic skeletal muscle stem cells. Stem Cell Reports 10:1398–1411. https://doi.org/10.1016/j.stemcr.2018.03.009

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Mauro A (1961) Satellite cell of skeletal muscle fibers. J Biophys Biochem Cytol 9:493–495

    Article  CAS  Google Scholar 

  5. Seale P, Sabourin LA, Girgis-Gabardo A et al (2000) Pax7 is required for the specification of myogenic satellite cells. Cell 102:777–786

    Article  CAS  Google Scholar 

  6. Seale P, Ishibashi J, Scimè A et al (2004) Pax7 is necessary and sufficient for the myogenic specification of CD45+:Sca1+ stem cells from injured muscle. PLoS Biol 2:E130

    Article  Google Scholar 

  7. Zanou N, Gailly P (2013) Skeletal muscle hypertrophy and regeneration: interplay between the myogenic regulatory factors (MRFs) and insulin-like growth factors (IGFs) pathways. Cell Mol Life Sci 70:4117–4130

    Article  CAS  Google Scholar 

  8. Hernandez-Torres F, Rodríguez-Outeiriño L, Franco D et al (2017) Pitx2 in embryonic and adult myogenesis. Front Cell Dev Biol 5:46

    Article  Google Scholar 

  9. Rando TA, Blau HM (1994) Primary mouse myoblast purification, characterization, and transplantation for cell-mediated gene therapy. J Cell Biol 125:1275–1287

    Article  CAS  Google Scholar 

  10. Hindi L, McMillan JD, Afroze D et al (2017) Isolation, culturing, and differentiation of primary myoblasts from skeletal muscle of adult mice. Bio Protoc 7

    Google Scholar 

  11. Shahini A, Vydiam K, Choudhury D et al (2018) Efficient and high yield isolation of myoblasts from skeletal muscle. Stem Cell Res 30:122–129

    Article  CAS  Google Scholar 

  12. Collins CA, Olsen I, Zammit PS et al (2005) Stem cell function, self-renewal, and behavioral heterogeneity of cells from the adult muscle satellite cell niche. Cell 122:289–301

    Article  CAS  Google Scholar 

  13. Montarras D, Morgan J, Collins C et al (2005) Direct isolation of satellite cells for skeletal muscle regeneration. Science 309:2064–2067

    Article  CAS  Google Scholar 

  14. Sacco A, Doyonnas R, Kraft P et al (2008) Self-renewal and expansion of single transplanted muscle stem cells. Nature 456:502–506

    Article  CAS  Google Scholar 

  15. Conboy MJ, Cerletti M, Wagers AJ et al (2010) Immuno-analysis and FACS sorting of adult muscle fiber-associated stem/precursor cells. Methods Mol Biol 621:165–173

    Article  CAS  Google Scholar 

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Correspondence to Amelia Aránega .

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Hernández-Torres, F., Rodríguez-Outeiriño, L., Aránega, A. (2020). Isolation and Culture of Quiescent Skeletal Muscle Satellite Cells. In: Kioussi, C. (eds) Stem Cells and Tissue Repair . Methods in Molecular Biology, vol 2155. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0655-1_12

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  • DOI: https://doi.org/10.1007/978-1-0716-0655-1_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0654-4

  • Online ISBN: 978-1-0716-0655-1

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