Division-Independent Differentiation of Muscle Stem Cells During a Growth Stimulus

Adult muscle stem cell (MuSC) behavior has been predominantly studied in the context of regeneration, a condition under which MuSCs exit quiescence and replicate upon activation before differentiating and fusing into myocytes. We use single-cell RNA-seq (scRNA-seq) to identify transcriptionally diverse subpopulations of MuSCs after 5 days of a growth stimulus. Overall design: MuSCs from plantaris muscles of doxyxycline treated Pax7rtTA; TRE-H2B-GFP mice, subjected to either sham surgery or 5-days of a growth stimulus, were isolated by Fluorescence-activated cell sorting (FACS) according to the prescence of GFP signal and analyzed using scRNA-seq.

成年肌肉干细胞（Adult muscle stem cell, MuSC）的生物学行为此前主要在再生情境中得以研究：在此过程中，MuSC会退出静息状态，在激活后发生增殖，随后分化并融合为肌细胞（myocytes）。本研究采用单细胞RNA测序（single-cell RNA-seq, scRNA-seq）技术，对经5天生长刺激处理的MuSCs的转录异质性亚群进行鉴定。实验整体设计：从经多西环素处理的Pax7rtTA；TRE-H2B-GFP双转基因小鼠的跖肌中分离得到MuSCs，将其分为假手术组与5天生长刺激组，随后根据绿色荧光蛋白信号的表达情况，通过荧光激活细胞分选（Fluorescence-activated cell sorting, FACS）分离目标细胞，并采用scRNA-seq完成测序分析。