FACS-based Satellite Cell Isolation From Mouse Hind Limb Muscles.
Fluorescence Activated Cell Sorting (FACS) is a sensitive and accurate method for purifying satellite cells, or muscle stem cells, from adult mouse skeletal muscle (Liuet al., 2013; Saccoet al., 2008; Tierneyet al., 2014). Mechanical and enzymatic digestion of hind limb muscles releases mononuclear muscle cells into suspension. This protocol employs fractionation strategies to deplete cells expressing the cell surface markers CD45, CD31, CD11b and Ly-6A/E-Sca1, both by magnetic separation and FACS-based exclusion, and positively select for cells expressing a7-integrin and CD34. This enables the researcher to successfully enrich satellite cells that uniformly express the paired-box transcription factor Pax7 and are capable of long-term self-renewal, skeletal muscle repair and muscle stem cell pool repopulation.
Development, Aging and Regeneration Program, Sanford-Burnham Medical Research Institute, La Jolla, USA.
Anastasia
A
Gromova
Development, Aging and Regeneration Program, Sanford-Burnham Medical Research Institute, La Jolla, USA; Graduate School of Biomedical Sciences, Sanford-Burnham Medical Research Institute, La Jolla, USA.
Matthew T
MT
Tierney
Development, Aging and Regeneration Program, Sanford-Burnham Medical Research Institute, La Jolla, USA.
Alessandra
A
Sacco
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