Skip to main content

Template DNA-Strand Co-Segregation and Asymmetric Cell Division in Skeletal Muscle Stem Cells

  • Protocol

Part of the book series: Methods in Molecular Biology ((MIMB,volume 482))

Abstract

Stem cells are present in all tissues and organs, and are crucial for normal regulated growth. How the pool size of stem cells and their progeny is regulated to establish the tissue prenatally, then maintain it throughout life, is a key question in biology and medicine. The ability to precisely locate stem and progenitors requires defining lineage progression from stem to differentiated cells, assessing the mode of cell expansion and self-renewal and identifying markers to assess the different cell states within the lineage. We have shown that during lineage progression from a quiescent adult muscle satellite cell to a differentiated myofibre, both symmetric and asymmetric divisions take place. Furthermore, we provide evidence that a sub-population of label retaining satellite cells co-segregate template DNA strands to one daughter cell. These findings provide a means of identifying presumed stem and progenitor cells within the lineage. In addition, asymmetric segregation of template DNA and the cytoplasmic protein Numb provides a landmark to define cell behaviour as self-renewal and differentiation decisions are being executed.

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

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
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

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Zammit, P.S., Partridge, T.A., Yablonka-Reuveni, Z. (2006) The skeletal muscle satellite cell: the stem cell that came in from the cold. J Histochem Cytochem 54, 1177–1191.

    Article  CAS  PubMed  Google Scholar 

  2. Tajbakhsh, S. (2005) keletal muscle stem and progenitor cells: reconciling genetics and lineage. Exp Cell Res 306, 364–372.

    Article  CAS  PubMed  Google Scholar 

  3. Morrison, S.J., Kimble, J. (2006) Asymmetric and symmetric stem-cell divisions in development and cancer. Nature 441, 1068–1074.

    Article  CAS  PubMed  Google Scholar 

  4. Shinin, V., Gayraud-Morel, B., Gomes, D., and Tajbakhsh, S. (2006) Asymmetric division and cosegregation of template DNA strands in adult muscle satellite cells. Nat Cell Biol 8, 677–682.

    Article  CAS  PubMed  Google Scholar 

  5. Cairns, J. (1975) Mutation selection and the natural history of cancer. Nature 255, 197–200.

    Article  CAS  PubMed  Google Scholar 

  6. Cairns, J. (2002) Somatic stem cells and the kinetics of mutagenesis and carcinogenesis. Proc Natl Acad Sci U S A 99, 10567–10570.

    Article  CAS  PubMed  Google Scholar 

  7. Mack, M., Riethmuller, G., Kufer, P. (1995) A small bispecific antibody construct expressed as a functional single-chain molecule with high tumor cell cytotoxicity. Proc Natl Acad Sci U S A 92, 7021–7025.

    Article  CAS  PubMed  Google Scholar 

  8. Campbell, R.E., Tour, O., Palmer, A.E., Steinbach, P. A., Baird, G. S., Zacharias, D. A., and Tsien, R. Y. (2002) A monomeric red fluorescent protein. Proc Natl Acad Sci U S A 99, 7877–7882.

    Article  CAS  PubMed  Google Scholar 

  9. Beauchamp, J.R., Heslop, L., Yu, D. S., Tajbakhsh, S., Kelly, R. G., Wernig, A., Buckingham, M. E., Partridge, T. A., and Zammit, P. S. (2000) Expression of CD34 and Myf5 defines the majority of quiescent adult skeletal muscle satellite cells. J Cell Biol 151, 1221–1234.

    Article  CAS  PubMed  Google Scholar 

  10. Tajbakhsh, S., Rocancourt, D., Cossu, G., and Buckingham, M. (1997) Redefining the genetic hierarchies controlling skeletal myogenesis: Pax-3 and Myf-5 act upstream of MyoD. Cell 89, 127–138.

    Article  CAS  PubMed  Google Scholar 

  11. Kassar-Duchossoy, L., Gayraud-Morel, B., Gomès, D., Rocancourt, D., Buckingham, M., Shinin, V., and Tajbakhsh, S. (2004) Mrf4 determines skeletal muscle identity in Myf5:Myod double-mutant mice. Nature 431, 466–471.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2009 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Shinin, V., Gayraud-Morel, B., Tajbakhsh, S. (2009). Template DNA-Strand Co-Segregation and Asymmetric Cell Division in Skeletal Muscle Stem Cells. In: Audet, J., Stanford, W.L. (eds) Stem Cells in Regenerative Medicine. Methods in Molecular Biology, vol 482. Humana Press. https://doi.org/10.1007/978-1-59745-060-7_19

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-060-7_19

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-797-6

  • Online ISBN: 978-1-59745-060-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics