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

Muscle stem cells (MuSCs) are the building blocks for regenerating skeletal mus-cle 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 mechan-ical dissociation of skeletal muscle coupled with fluorescence-activated cell sort-ing (FACS). FACS-isolated MuSCs can be used for various downstream applica-tions 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

Automated summary

This article presents a protocol for isolating mouse muscle stem cells (MuSCs) using a two-step enzymatic and mechanical dissociation method coupled with fluorescence-activated cell sorting (FACS). The FACS-isolated MuSCs can be utilized for various downstream applications, including cell culture, cell transduction, immunofluorescence, and gene expression assays.