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Protocol for Isolation and Characterization of In Situ Fixed Quiescent Muscle Stem Cells.

ABSTRACT: Quiescent muscle stem cells, also called satellite cells (SCs), are essential for muscle regeneration. However, quiescent SCs are quickly activated during fluorescence-activated cell sorting (FACS) isolation. Here, we present an optimized protocol to isolate quiescent muscle stem cells from fixative-perfused mice and generate high-quality cDNA libraries for RNA-sequencing analysis. Fixation preserves the signatures of quiescent muscle stem cells in vivo. Isolated SCs can be used for downstream analysis such as immunofluorescence, RNA sequencing, and mass spectrometry. For complete information on the use and execution of this protocol, please refer to Yue et al. (2020).

SUBMITTER: Yue L

PROVIDER: S-EPMC7757111 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Protocol for Isolation and Characterization of <i>In Situ</i> Fixed Quiescent Muscle Stem Cells.

Yue Lu L Cheung Tom H TH

STAR protocols 20201009 3

Quiescent muscle stem cells, also called satellite cells (SCs), are essential for muscle regeneration. However, quiescent SCs are quickly activated during fluorescence-activated cell sorting (FACS) isolation. Here, we present an optimized protocol to isolate quiescent muscle stem cells from fixative-perfused mice and generate high-quality cDNA libraries for RNA-sequencing analysis. Fixation preserves the signatures of quiescent muscle stem cells <i>in vivo</i>. Isolated SCs can be used for downs ...[more]

PMID: 33377022

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