Duchenne muscular dystrophy (DMD) is a progressive and fatal muscle-wasting disease caused by DYSTROPHIN deficiency. Cell therapy using muscle stem cells (MuSCs) is a potential cure. Here, we report a differentiation method to generate fetal MuSCs from human induced pluripotent stem cells (iPSCs) by monitoring MYF5 expression. Gene expression profiling indicated that MYF5-positive cells in the late stage of differentiation have fetal MuSC characteristics, while MYF5-positive cells in the early stage of differentiation have early myogenic progenitor characteristics. Moreover, late-stage MYF5-positive cells demonstrated good muscle regeneration potential and produced DYSTROPHIN in vivo after transplantation into DMD model mice, resulting in muscle function recovery. The engrafted cells also generated PAX7-positive MuSC-like cells under the basal lamina of DYSTROPHIN-positive fibers. These findings suggest that MYF5-positive fetal MuSCs induced in the late stage of iPSC differentiation have cell therapy potential for DMD.

Duchenne肌营养不良症（DMD）是由DYSTROPHIN缺乏症引起的进行性和致命性肌肉萎缩性疾病。使用肌肉干细胞（MuSC）的细胞疗法是一种潜在的治疗方法。在这里，我们报告了一种通过监测MYF5表达从人诱导的多能干细胞（iPSCs）生成胎儿MuSCs的分化方法。基因表达谱分析表明，分化晚期的MYF5阳性细胞具有胎儿MuSC特征，而分化早期的MYF5阳性细胞具有早期成肌祖细胞特征。此外，晚期MYF5阳性细胞显示出良好的肌肉再生潜能，并在体内产生了DYSTROPHIN移植到DMD模型小鼠后，导致肌肉功能恢复。植入的细胞还在DYSTROPHIN阳性纤维的基底层下产生PAX7阳性MuSC样细胞。这些发现表明，在iPSC分化后期诱导的MYF5阳性胎儿MuSC具有DMD的细胞治疗潜力。