Skip to main content

Isolation of Skeletal Muscle Stem Cells for Phenotypic Screens for Modulators of Proliferation

  • Protocol
  • First Online:
Phenotypic Screening

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1787))

Abstract

Adult skeletal muscle contains a population of resident stem cells known as muscle stem cells (MuSC) or satellite cells. This population of cells is required for regeneration of functional myofibers after damage. Aging reduces the proliferative response of satellite cells post-injury. This deficient response is thought to contribute to slowed recovery of muscle function after damage in the elderly and may also contribute to age-related loss of muscle function (sarcopenia). Numerous techniques are now available for the isolation of highly purified satellite cells from mice and humans (Sherwood, et al. Cell 119:543–554, 2004; Cerletti, et al. Cell 134:37–47; 2008; Conboy, et al. Methods Mol Biol 621:165–173, 2010; Bareja, et al. PLoS One 9:e90398; 2014; Castiglioni et al. Stem Cell Rep 2:92–106, 2014; Charville, et al. Stem Cell Rep 5:621–632, 2015; Liu et al. Nat Protoc 10:1612–1624, 2015; Sincennes et al. Methods Mol Biol 1556:41–50, 2017), thus opening an opportunity to use satellite cells in phenotypic screens for regulators of satellite cell proliferation and differentiation. In this chapter, we describe a technique for the prospective isolation of mouse satellite cells that we have recently used in a phenotypic screen of a focused set of small molecules.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Sherwood RI, Christensen JL, Conboy IM, Conboy MJ, Rando TA, Weissman IL et al (2004) Isolation of adult mouse myogenic progenitors: functional heterogeneity of cells within and engrafting skeletal muscle. Cell 119:543–554

    Article  CAS  Google Scholar 

  2. Cerletti M, Jurga S, Witczak CA, Hirshman MF, Shadrach JL, Goodyear LJ et al (2008) Highly efficient, functional engraftment of skeletal muscle stem cells in dystrophic muscles. Cell 134:37–47

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  4. Bareja A, Holt JA, Luo G, Chang C, Lin J, Hinken AC et al (2014) Human and mouse skeletal muscle stem cells: convergent and divergent mechanisms of myogenesis. PLoS One 9:e90398

    Article  Google Scholar 

  5. Castiglioni A, Hettmer S, Lynes MD, Rao TN, Tchessalova D, Sinha I et al (2014) Isolation of progenitors that exhibit myogenic/osteogenic bipotency in vitro by fluorescence-activated cell sorting from human fetal muscle. Stem Cell Rep 2:92–106

    Article  CAS  Google Scholar 

  6. Charville GW, Cheung TH, Yoo B, Santos PJ, Lee GK, Shrager JB et al (2015) Ex vivo expansion and in vivo self-renewal of human muscle stem cells. Stem Cell Rep 5:621–632

    Article  CAS  Google Scholar 

  7. Liu L, Cheung TH, Charville GW, Rando TA (2015) Isolation of skeletal muscle stem cells by fluorescence-activated cell sorting. Nat Protoc 10:1612–1624

    Article  CAS  Google Scholar 

  8. Sincennes MC, Wang YX, Rudnicki MA (2017) Primary mouse myoblast purification using magnetic cell separation. Methods Mol Biol 1556:41–50

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  10. Billin AN, Bantscheff M, Drewes G, Ghidelli-Disse S, Holt JA, Kramer HF et al (2016) Discovery of novel small molecules that activate satellite cell proliferation and enhance repair of damaged muscle. ACS Chem Biol 11:518–529

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Aaron C. Hinken .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Hinken, A.C., Billin, A.N. (2018). Isolation of Skeletal Muscle Stem Cells for Phenotypic Screens for Modulators of Proliferation. In: Wagner, B. (eds) Phenotypic Screening. Methods in Molecular Biology, vol 1787. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7847-2_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7847-2_6

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7846-5

  • Online ISBN: 978-1-4939-7847-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics