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Purification of Myogenic Progenitors from Human Muscle Using Fluorescence-Activated Cell Sorting (FACS)

  • Anna Pakula
  • Janelle M. Spinazzola
  • Emanuela GussoniEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1889)

Abstract

Primary myoblasts derived from human tissue are a valuable tool in research of muscle disease and pathophysiology. However, skeletal muscle biopsies, especially from diseased muscle, contain a plethora of non-myogenic cells, necessitating purification of the myogenic cell population. This protocol describes techniques for dissociation of cells from human skeletal muscle biopsies and enrichment for a highly myogenic population by fluorescence-activated cell sorting (FACS). We also describe methods for assessing myogenicity and population expansion for subsequent in vitro study.

Key words

Skeletal muscle Myoblast isolation Tissue dissociation Fluorescence-activated cell sorting (FACS) CD82 CD56 Immunostaining Pax7 

Notes

Acknowledgments

This work is supported by a grant from the Muscular Dystrophy Association #479606 (EG) and by the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health under Award Number 1R01AR069582-01 (EG). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. This protocol was modified from previous work, specifically from the listed references [10, 11].

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Anna Pakula
    • 1
    • 2
  • Janelle M. Spinazzola
    • 1
    • 2
  • Emanuela Gussoni
    • 1
    • 2
    • 3
    Email author
  1. 1.Division of Genetics and GenomicsBoston Children’s HospitalBostonUSA
  2. 2.Department of PediatricsHarvard Medical SchoolBostonUSA
  3. 3.The Stem Cell Program at Boston Children’s HospitalBostonUSA

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