流式细胞仪分析、分离和染色小鼠肌肉卫星细胞。

Flow Cytometer Analyses, Isolation, and Staining of Murine Muscle Satellite Cells.

机构信息

Laboratory of Stem Cell Regeneration and Adaptation, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.

出版信息

Methods Mol Biol. 2023;2640:3-11. doi: 10.1007/978-1-0716-3036-5_1.

DOI:10.1007/978-1-0716-3036-5_1

PMID:36995583

Abstract

Fluorescence-activated cell sorting (FACS) is a powerful and requisite tool for the analysis and purification of adult stem cells. However, it is difficult to separate adult stem cells from solid organs than from immune-related tissues/organs. This is because of the presence of large amounts of debris, which increases noise in the FACS profiles. In particular, it is extremely difficult for unfamiliar researchers to identify muscle stem cell (also known as muscle satellite cell: MuSC) fraction because all myofibers, which are mainly composed of skeletal muscle tissues, become debris during cell preparation. This chapter describes our FACS protocol, which we have used for more than a decade, to identify and purify MuSCs.

摘要

荧光激活细胞分选（FACS）是分析和纯化成体干细胞的有力且必需的工具。然而，从实体器官中分离成体干细胞比从免疫相关组织/器官中分离要困难。这是因为存在大量的碎片，这会增加 FACS 图谱中的噪声。特别是，对于不熟悉的研究人员来说，识别肌肉干细胞（也称为肌肉卫星细胞：MuSC）部分非常困难，因为在细胞制备过程中，所有主要由骨骼肌组织组成的肌纤维都会变成碎片。本章描述了我们十年来一直使用的 FACS 方案，用于鉴定和纯化 MuSCs。