Protocol for the isolation of mouse muscle stem cells using fluorescence-activated cell sorting

Gabriel Elizalde; Alma Zuniga Munoz; Albert E Almada

doi:10.1016/j.xpro.2023.102656

Protocol for the isolation of mouse muscle stem cells using fluorescence-activated cell sorting

STAR Protoc. 2023 Dec 15;4(4):102656. doi: 10.1016/j.xpro.2023.102656. Epub 2023 Oct 22.

Authors

Gabriel Elizalde¹, Alma Zuniga Munoz¹, Albert E Almada²

Affiliations

¹ Department of Orthopaedic Surgery, University of Southern California, Los Angeles, CA 90033, USA; Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles, CA 90033, USA.
² Department of Orthopaedic Surgery, University of Southern California, Los Angeles, CA 90033, USA; Department of Stem Cell Biology and Regenerative Medicine, University of Southern California, Los Angeles, CA 90033, USA. Electronic address: [email protected].

Abstract

Muscle stem cells (MuSCs) are the building blocks for regenerating skeletal muscle after trauma. If we intend to maximize the therapeutic potential of MuSCs, we must further study their molecular and functional properties. Here, we present a protocol for the isolation of mouse MuSCs via a two-step enzymatic and mechanical dissociation of skeletal muscle coupled with fluorescence-activated cell sorting (FACS). FACS-isolated MuSCs can be used for various downstream applications including cell culture, cell transduction, immunofluorescence, and gene expression assays. For complete details on the use and execution of this protocol, please refer to Almada et al. (2021).¹.

Keywords: Antibody; Cell Differentiation; Cell isolation; Flow Cytometry; Gene Expression; Stem Cells.

MeSH terms

Animals
Cell Culture Techniques
Flow Cytometry / methods
Mice
Muscle, Skeletal*
Satellite Cells, Skeletal Muscle*
Stem Cells

Grants and funding

R01 AR080753/AR/NIAMS NIH HHS/United States