Advertisement

pp 1-9 | Cite as

Surface Antigen-Based Identification of In Vitro Expanded Skeletal Muscle-Derived Mesenchymal Stromal/Stem Cells Using Flow Cytometry

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

Abstract

Mesenchymal stem/stromal cells (MSCs) can be isolated from several connective tissues in the adult organism by harnessing their propensity for plastic adherence in vitro. Upon culture expansion, the resulting cell cultures are composed of many different cell types at different stages of differentiation. Hence, their identity must be confirmed. Flow cytometry is an indispensable method for accurate quantification of MSC surface antigens. Here, we present a protocol that uses flow cytometry for the identification of MSCs based on the set of surface antigens required by the International Society for Cellular Therapy.

Keywords

Flow cytometry Identification Mesenchymal stem/stromal cells Skeletal muscle Surface markers 

Notes

Acknowledgements

This work was supported by the Slovenian Research Agency, J3-7245 Research Project and P3-0298 Research Programme.

References

  1. 1.
    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/s12015-018-9800-6CrossRefGoogle Scholar
  2. 2.
    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.069088CrossRefGoogle Scholar
  3. 3.
    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.2016.00069CrossRefGoogle Scholar
  4. 4.
    Dominici M, Le BK, Mueller I et al (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8:315–317Google Scholar
  5. 5.
    Hawley TS, Hawley RG (eds) (2018) Flow cytometry protocols. Springer, New York, NYGoogle Scholar
  6. 6.
    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 2:112–120Google Scholar

Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

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

Personalised recommendations