PDF(Pubmed)
Abstract：
Satellite cells are myogenic cells responsible for muscle growth shortly after birth and muscle repair/regeneration during adulthood. Therapies based on satellite cells hold promise for treating muscular dysfunctions. Studying satellite cells is technically challenging owing to their low abundance, small size and anatomical dispersed location between the basal lamina and the sarcolemma of myofibers. In this article, we present three improved protocol strategies for studying the properties of satellite cells of the mouse during the different stages of muscle regeneration: (1) immunostaining of freshly isolated single myofibers to facilitate the study of quiescent satellite cells, (2) cultivation of single myofibers on Matrigel®-coated dish to study the myogenesis programs initiated by satellite cell activation, and (3) cultivation of single myofibers in floating conditions to analyze activated satellite cells or the doubling time of satellite cells in myofibers. In brief, when compared to previously published protocols, this article presented an improved protocol that requires shorter experimental time and less laborious approach for higher yield of intact single myofibers for downstream analyses.
摘要：
卫星细胞是在出生后不久负责肌肉生长和成年期肌肉修复/再生的生肌细胞。基于卫星细胞的疗法有望治疗肌肉功能障碍。由于卫星细胞的丰度低，研究卫星细胞在技术上具有挑战性，小尺寸和解剖分散的位置之间的基底层和肌纤维的肌膜。在这篇文章中,我们提出了三种改进的方案策略，用于研究小鼠在肌肉再生的不同阶段的卫星细胞的特性：（1）新分离的单个肌纤维的免疫染色，以促进静止卫星细胞的研究，（2）在Matrigel®包被的培养皿上培养单肌纤维，以研究由卫星细胞活化启动的成肌程序，（3）在漂浮条件下培养单个肌纤维，以分析激活的卫星细胞或卫星细胞在肌纤维中的倍增时间。简而言之,与以前发表的协议相比，本文提出了一种改进的方案，该方案需要更短的实验时间和更少的费力方法，以获得更高的完整单肌纤维用于下游分析。
