Human pluripotent stem cells have the potential to differentiate into various cell types including skeletal muscles (SkM), and they are applied to regenerative medicine or in vitro modelling for intractable diseases. A simple differentiation method is required for SkM cells to accelerate neuromuscular disease studies. Here, we established a simple method to convert human pluripotent stem cells into SkM cells by using temperature-sensitive Sendai virus (SeV) vector encoding myoblast determination protein 1 (SeV-Myod1), a myogenic master transcription factor. SeV-Myod1 treatment converted human embryonic stem cells (ESCs) into SkM cells, which expressed SkM markers including myosin heavy chain (MHC). We then removed the SeV vector by temporal treatment at a high temperature of 38℃, which also accelerated mesodermal differentiation, and found that SkM cells exhibited fibre-like morphology. Finally, after removal of the residual human ESCs by pluripotent stem cell-targeting delivery of cytotoxic compound, we generated SkM cells with 80% MHC positivity and responsiveness to electrical stimulation. This simple method for myogenic differentiation was applicable to human-induced pluripotent stem cells and will be beneficial for investigations of disease mechanisms and drug discovery in the future.

中文翻译：

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利用温度敏感的仙台病毒载体从人多能干细胞中简单衍生骨骼肌

人类多能干细胞具有分化成包括骨骼肌 (SkM) 在内的各种细胞类型的潜力，它们被应用于再生医学或难治性疾病的体外建模。SkM 细胞需要一种简单的分化方法来加速神经肌肉疾病的研究。在这里，我们建立了一种简单的方法，通过使用编码成肌细胞决定蛋白 1 (SeV-Myod1) 的温度敏感仙台病毒 (SeV) 载体将人类多能干细胞转化为 SkM 细胞，这是一种生肌主转录因子。SeV-Myod1 处理将人类胚胎干细胞 (ESC) 转化为 SkM 细胞，表达包括肌球蛋白重链 (MHC) 在内的 SkM 标记。然后我们在38℃的高温下进行临时处理去除了SeV载体，这也加速了中胚层分化，并发现SkM细胞表现出纤维状形态。最后，在通过多能干细胞靶向递送细胞毒性化合物去除残留的人类胚胎干细胞后，我们产生了具有 80% MHC 阳性和对电刺激反应的 SkM 细胞。这种简单的肌原性分化方法适用于人类诱导的多能干细胞，将有利于未来疾病机制的研究和药物发现。