Protocol for the isolation of mouse muscle stem cells using fluorescence-activated cell sorting.
Publication Year:
2023
PubMed ID:
37874680
Funding Grants:
Public Summary:
Muscle stem cells (MuSCs) help rebuild damaged muscle after injury, making them valuable for muscle therapies. This protocol describes a method to isolate MuSCs from mice using a two-step process: breaking down muscle tissue with enzymes and then separating the cells with a technique called fluorescence-activated cell sorting (FACS). Once isolated, these MuSCs can be used in various lab studies, like growing them in culture, studying their genes, or tracking them with fluorescent markers for further research.
Scientific 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).
