Gallot Yann Simon, Hindi Sajedah M, Mann Aman K, Kumar Ashok
Department of Anatomical Sciences and Neurobiology, University of Louisville School of Medicine, Louisville, USA.
duPont Manual High School, Louisville, USA.
Bio Protoc. 2016 Oct 5;6(19). doi: 10.21769/BioProtoc.1942.
Adult skeletal muscle regeneration is orchestrated by a specialized population of adult stem cells called satellite cells, which are localized between the basal lamina and the plasma membrane of myofibers. The process of satellite cell-activation, proliferation, and subsequent differentiation that occurs during muscle regeneration can be recapitulated by isolation of single myofibers from skeletal muscles and culturing them under suspension conditions. Here, we describe an improved protocol to evaluate satellite cells activation through isolation of single myofibers from extensor digitorum longus (EDL) muscle of mice and culturing and staining of myofiber-associated satellite cells with the markers of self-renewal, proliferation, and differentiation.
成体骨骼肌的再生由一种称为卫星细胞的成体干细胞特殊群体精心调控，这些卫星细胞位于肌纤维的基膜和质膜之间。在肌肉再生过程中发生的卫星细胞激活、增殖及随后的分化过程，可以通过从骨骼肌中分离单个肌纤维并在悬浮条件下培养来重现。在此，我们描述了一种改进的方案，通过从小鼠趾长伸肌（EDL）中分离单个肌纤维，并对与肌纤维相关的卫星细胞进行培养和用自我更新、增殖及分化标记物染色，来评估卫星细胞的激活情况。
