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Human Skeletal Muscle-Derived Mesenchymal Stem/Stromal Cell Isolation and Growth Kinetics Analysis

  • Klemen Čamernik
  • Janja Marc
  • Janja ZupanEmail author
Part of the Methods in Molecular Biology book series


The most studied sources of mesenchymal stem/stromal cells (MSCs) are bone marrow and adipose tissue. However skeletal muscle represents an interesting source of diverse subpopulations of MSCs, such as paired box 7 (Pax-7)-positive satellite cells, fibro-/adipogenic progenitors, PW1-positive interstitial cells and others. The specific properties of some of these muscle-derived cells have encouraged the development of cell therapies for muscle regeneration. However, the identity and multilineage potential of the diverse muscle-resident cells should first be evaluated in vitro, followed by in vivo clinical trials to predict their regenerative capacity. Here, we present protocols for the isolation of MSCs from skeletal muscle using enzymatic digestion and mechanical trituration. We also provide a method to determine their specific growth rate, a feature that is of particular interest when designing cell therapies.


Cell growth kinetics Collagenase digestion Isolation Mesenchymal stem/stromal cells Skeletal muscle 



This work was supported by the Slovenian Research Agency, J3-7245 Research Project and P3-0298 Research Programme and by the ARTE Project EU Interreg Italia Slovenia 2014-2020.


  1. 1.
    Horwitz EM, Le BK, Dominici M et al (2005) Clarification of the nomenclature for MSCs: the International Society for Cellular Therapy position statement. Cytotherapy 7:393–395Google Scholar
  2. 2.
    Friedenstein AJ, Gorskaja JF, Kulagina NN (1976) Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 4:267–274Google Scholar
  3. 3.
    Čamernik K, Barlič A, Drobnič M et al (2018) Mesenchymal stem cells in the musculoskeletal system: from animal models to human tissue regeneration? Stem Cell Rev 14(3):346–369. Scholar
  4. 4.
    Relaix F, Zammit PS (2012) Satellite cells are essential for skeletal muscle regeneration: the cell on the edge returns centre stage. Development 139:2845–2856. Scholar
  5. 5.
    Hamrick MW, McGee-Lawrence ME, Frechette DM (2016) Fatty infiltration of skeletal muscle: mechanisms and comparisons with bone marrow adiposity. Front Endocrinol (Lausanne) 7:1–7. Scholar
  6. 6.
    Cottle BJ, Lewis FC, Shone V et al (2017) Skeletal muscle-derived interstitial progenitor cells (PICs) display stem cell properties, being clonogenic, self-renewing, and multi-potent in vitro and in vivo. Stem Cell Res Ther 8(1):158. Scholar
  7. 7.
    Mizukami A, Swiech K (2018) Mesenchymal stromal cells: from discovery to manufacturing and commercialization. Stem Cells Int 2018:4083921. Scholar
  8. 8.
    Heathman TRJ, Rafiq QA, Chan AKC et al (2016) Characterization of human mesenchymal stem cells from multiple donors and the implications for large-scale bioprocess development. Biochem Eng J 108:14–23. Scholar
  9. 9.
    Qi W, Yuan W, Yan J et al (2014) Growth and accelerated differentiation of mesenchymal stem cells on graphene oxide/poly-l-lysine composite films. J Mater Chem B 2:5461–5467. Scholar
  10. 10.
    Tsai C-C, Yew T-L, Yang D-C et al (2012) Benefits of hypoxic culture on bone marrow multipotent stromal cells. Am J Blood Res 2:148–159Google Scholar
  11. 11.
    Autengruber A, Gereke M, Hansen G et al (2012) Impact of enzymatic tissue disintegration on the level of surface molecule expression and immune cell function. Eur J Microbiol Immunol 2:112–120. Scholar

Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  1. 1.Faculty of Pharmacy, Department of Clinical BiochemistryUniversity of LjubljanaLjubljanaSlovenia

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