Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Isolation of myogenic progenitor populations from Pax7-deficient skeletal muscle based on adhesion characteristics

Gene Therapy volume 15, pages 1116–1125 (2008)Cite this article

Abstract

In an attempt to determine whether muscle-derived stem cells are distinct from satellite cells, we investigated whether muscle-derived stem cells could be isolated from the skeletal muscle of Pax7-deficient mice, which have been shown to be devoid of or to contain only a minimal number of satellite cells. Utilizing a technique that separates cells based on their adhesion characteristics (the preplate technique), several distinct populations of muscle-derived cells were isolated. In these mice, the Pax7 gene was knocked out with the insertion of the LacZ gene. One population was both rapidly adhering, LacZ-positive, and displayed a high myogenic index, but was rapidly lost to terminal differentiation when continuously replated. A second population, which persisted over 50 passages, was LacZ-negative and displayed a low myogenic index. Although Pax3 may have acted as a compensatory mechanism for the myogenic commitment of the LacZ-positive cells, the LacZ-negative cells, despite expressing Pax3, required Pax7 transduction to restore their myogenic capacity. We believe that these two populations of myogenic progenitor cells, each endowed with different adhesion characteristics, may help explain the discrepancy in the literature concerning the presence of myogenic cells found in Pax7-deficient mice.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Charge SB, Rudnicki MA . Cellular and molecular regulation of muscle regeneration. Physiol Rev 2004; 84: 209–238.

    Article  CAS  Google Scholar 

  2. Deasy BM, Gharaibeh BM, Pollett JB, Jones MM, Lucas MA, Kanda Y et al. Long-term self-renewal of postnatal muscle-derived stem cells. Mol Biol Cell 2005; 16: 3323–3333.

    Article  CAS  Google Scholar 

  3. Qu-Petersen Z, Deasy B, Jankowski R, Ikezawa M, Cummins J, Pruchnic R et al. Identification of a novel population of muscle stem cells in mice: potential for muscle regeneration. J Cell Biol 2002; 157: 851–864.

    Article  CAS  Google Scholar 

  4. Seale P, Sabourin LA, Girgis-Gabardo A, Mansouri A, Gruss P, Rudnicki MA . Pax7 is required for the specification of myogenic satellite cells. Cell 2000; 102: 777–786.

    Article  CAS  Google Scholar 

  5. Mansouri A, Stoykova A, Torres M, Gruss P . Dysgenesis of cephalic neural crest derivatives in Pax7-/- mutant mice. Development 1996; 122: 831–838.

    CAS  PubMed  Google Scholar 

  6. Bober E, Franz T, Arnold HH, Gruss P, Tremblay P . Pax-3 is required for the development of limb muscles: a possible role for the migration of dermomyotomal muscle progenitor cells. Development 1994; 120: 603–612.

    CAS  PubMed  Google Scholar 

  7. Goulding M, Lumsden A, Paquette AJ . Regulation of Pax-3 expression in the dermomyotome and its role in muscle development. Development 1994; 120: 957–971.

    CAS  PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  9. Relaix F, Montarras D, Zaffran S, Gayraud-Morel B, Rocancourt D, Tajbakhsh S et al. Pax3 and Pax7 have distinct and overlapping functions in adult muscle progenitor cells. J Cell Biol 2006; 172: 91–102.

    Article  CAS  Google Scholar 

  10. Relaix F, Rocancourt D, Mansouri A, Buckingham M . Divergent functions of murine Pax3 and Pax7 in limb muscle development. Genes Dev 2004; 18: 1088–1105.

    Article  CAS  Google Scholar 

  11. Kuang S, Chargé SB, Seale P, Huh M, Rudnicki MA . Distinct roles for Pax7 and Pax3 in adult regenerative myogenesis. J Cell Biol 2006; 172: 103–113.

    Article  CAS  Google Scholar 

  12. Winitsky SO, Gopal TV, Hassanzadeh S, Takahashi H, Gryder D, Rogawski MA et al. Adult murine skeletal muscle contains cells that can differentiate into beating cardiomyocytes in vitro. PLoS Biol 2005; 3: e87.

    Article  Google Scholar 

  13. Sarig R, Baruchi Z, Fuchs O, Nudel U, Yaffe D . Regeneration and transdifferentiation potential of muscle-derived stem cells propagated as myospheres. Stem Cells 2006; 24: 1769–1778.

    Article  Google Scholar 

  14. Nomura T, Ashihara E, Tateishi K, Asada S, Ueyama T, Takahashi T et al. Skeletal myosphere-derived progenitor cell transplantation promotes neovascularization in delta-sarcoglycan knockdown cardiomyopathy. Biochem Biophys Res Commun 2007; 352: 668–674.

    Article  CAS  Google Scholar 

  15. Rouger K, Fornasari B, Armengol V, Jouvion G, Leroux I, Dubreil L et al. Progenitor cell isolation from muscle-derived cells based on adhesion properties. J Histochem Cytochem 2007; 55: 607–618.

    Article  CAS  Google Scholar 

  16. Spangrude GJ, Heimfeld S, Weissman IL . Purification and characterization of mouse hematopoietic stem cells. Science 1988; 241: 58–62.

    Article  CAS  Google Scholar 

  17. Tumbar T, Guasch G, Greco V, Blanpain C, Lowry WE, Rendl M et al. Defining the epithelial stem cell niche in skin. Science 2004; 303: 359–363.

    Article  CAS  Google Scholar 

  18. Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 2001; 7: 211–228.

    Article  CAS  Google Scholar 

  19. Péault B, Rudnicki M, Torrente Y, Cossu G, Tremblay JP, Partridge T et al. Stem and progenitor cells in skeletal muscle development, maintenance, and therapy. Mol Ther 2007; 15: 867–877.

    Article  Google Scholar 

  20. Richler C, Yaffe D . The in vitro cultivation and differentiation capacities of myogenic cell lines. Dev Biol 1970; 23: 1–22.

    Article  CAS  Google Scholar 

  21. Hwang JH, Yuk SH, Lee JH, Lyoo WS, Ghil SH, Lee SS et al. Isolation of muscle derived stem cells from rat and its smooth muscle differentiation [corrected]. Mol Cells 2004; 17: 57–61.

    CAS  PubMed  Google Scholar 

  22. Beauchamp JR, Heslop L, Yu DS, Tajbakhsh S, Kelly RG, Wernig A et al. Expression of CD34 and Myf5 defines the majority of quiescent adult skeletal muscle satellite cells. J Cell Biol 2000; 151: 1221–1234.

    Article  CAS  Google Scholar 

  23. Cornelison DD, Wold BJ . Single-cell analysis of regulatory gene expression in quiescent and activated mouse skeletal muscle satellite cells. Dev Biol 1997; 191: 270–283.

    Article  CAS  Google Scholar 

  24. Lee JY, Qu-Petersen Z, Cao B, Kimura S, Jankowski R, Cummins J et al. Clonal isolation of muscle-derived cells capable of enhancing muscle regeneration and bone healing. J Cell Biol 2000; 150: 1085–1100.

    Article  CAS  Google Scholar 

  25. Oustanina S, Hause G, Braun T . Pax7 directs postnatal renewal and propagation of myogenic satellite cells but not their specification. EMBO J 2004; 23: 3430–3439.

    Article  CAS  Google Scholar 

  26. Olguin HC, Olwin BB . Pax-7 up-regulation inhibits myogenesis and cell cycle progression in satellite cells: a potential mechanism for self-renewal. Dev Biol 2004; 275: 375–388.

    Article  CAS  Google Scholar 

  27. Zammit PS, Relaix F, Nagata Y, Ruiz AP, Collins CA, Partridge TA et al. Pax7 and myogenic progression in skeletal muscle satellite cells. J Cell Sci 2006; 119: 1824–1832.

    Article  CAS  Google Scholar 

  28. Jostes B, Walther C, Gruss P . The murine paired box gene, Pax7, is expressed specifically during the development of the nervous and muscular system. Mech Dev 1990; 33: 27–37.

    Article  CAS  Google Scholar 

  29. Snider L, Tapscott SJ . Emerging parallels in the generation and regeneration of skeletal muscle. Cell 2003; 113: 811–812.

    Article  CAS  Google Scholar 

  30. Qu Z, Huard J . Matching host muscle and donor myoblasts for myosin heavy chain improves myoblast transfer therapy. Gene Therapy 2000; 7: 428–437.

    Article  CAS  Google Scholar 

  31. Seale P, Ishibashi J, Scime A, Rudnicki MA . Pax7 is necessary and sufficient for the myogenic specification of CD45+:Sca1+ stem cells from injured muscle. PLoS Biol 2004; 2: E130.

    Article  Google Scholar 

  32. Graham FL, van der Eb AJ . A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 1973; 52: 456–467.

    Article  CAS  Google Scholar 

  33. Peng H, Wright V, Usas A, Gearhart B, Shen HC, Cummins J et al. Synergistic enhancement of bone formation and healing by stem cell-expressed VEGF and bone morphogenetic protein-4. J Clin Invest 2002; 110: 751–759.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Orthopaedic Surgery, Stem Cell Research Center, Children's Hospital of Pittsburgh, University of Pittsburgh, Pittsburgh, PA, USA

    A Lu, J H Cummins, J B Pollett, B Cao, B Sun & J Huard

  2. Department of Medicine, Ottawa Health Research Institute, University of Ottawa, Ottawa, Ontario, Canada

    M A Rudnicki

Authors
  1. A Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J H Cummins
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J B Pollett
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. B Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. B Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M A Rudnicki
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J Huard
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to J Huard.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lu, A., Cummins, J., Pollett, J. et al. Isolation of myogenic progenitor populations from Pax7-deficient skeletal muscle based on adhesion characteristics. Gene Ther 15, 1116–1125 (2008). https://doi.org/10.1038/gt.2008.86

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/gt.2008.86

Keywords

This article is cited by

Search

Advanced search

Quick links