Human Skeletal Muscle-Derived Mesenchymal Stem/Stromal Cell Isolation and Growth Kinetics Analysis Protocol First Online: 30 November 2018
Part of the
Methods in Molecular Biology
book series Abstract
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.
Keywords Cell growth kinetics Collagenase digestion Isolation Mesenchymal stem/stromal cells Skeletal muscle Notes Acknowledgements
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.
Horwitz EM, Le BK, Dominici M et al (2005) Clarification of the nomenclature for MSCs: the International Society for Cellular Therapy position statement. Cytotherapy 7:393–395
Friedenstein AJ, Gorskaja JF, Kulagina NN (1976) Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 4:267–274
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.
https://doi.org/10.1007/s12015-018-9800-6 CrossRef Google Scholar
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.
https://doi.org/10.1242/dev.069088 CrossRef Google Scholar
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.
https://doi.org/10.3389/fendo.2016.00069 CrossRef Google Scholar
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
. Stem Cell Res Ther 8(1):158.
https://doi.org/10.1186/s13287-017-0612-4 CrossRef Google Scholar
Mizukami A, Swiech K (2018) Mesenchymal stromal cells: from discovery to manufacturing and commercialization. Stem Cells Int 2018:4083921.
https://doi.org/10.1155/2018/4083921 CrossRef Google Scholar
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.
https://doi.org/10.1016/j.bej.2015.06.018 CrossRef Google Scholar
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.
https://doi.org/10.1039/C4TB00856A CrossRef Google Scholar
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 2:148–159
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.
https://doi.org/10.1556/EuJMI.2.2012.2.3 CrossRef Google Scholar Copyright information
© Springer Science+Business Media New York 2018