固定静息肌肉干细胞的分离和鉴定方案。

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

机构信息

Division of Life Science, Center for Stem Cell Research, Center of Systems Biology and Human Health, State Key Laboratory of Molecular Neuroscience, and Molecular Neuroscience Center, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China.

Hong Kong Center for Neurodegenerative Diseases, Hong Kong Science Park, Hong Kong, China.

出版信息

STAR Protoc. 2020 Oct 9;1(3):100128. doi: 10.1016/j.xpro.2020.100128. eCollection 2020 Dec 18.

DOI:10.1016/j.xpro.2020.100128

PMID:33377022

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7757111/

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 . 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).

摘要

静止的肌肉干细胞，也称为卫星细胞 (SCs)，是肌肉再生所必需的。然而，在荧光激活细胞分选 (FACS) 分离过程中，静止的 SC 会迅速被激活。在这里，我们提出了一种从固定灌注的小鼠中分离静止的肌肉干细胞的优化方案，并为 RNA-seq 分析生成高质量的 cDNA 文库。固定保存了静止的肌肉干细胞的特征。分离的 SC 可用于下游分析，如免疫荧光、RNA 测序和质谱分析。有关该方案使用和执行的完整信息，请参阅 Yue 等人 (2020)。