Generation of human muscle fibers and satellite-like cells from human pluripotent stem cells in vitro
Generation of human muscle fibers and satellite-like cells from human pluripotent stem cells in vitro
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DOI：
10.1038/nprot.2016.110
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发表时间：
2016-10-01
期刊：
影响因子：
14.8
通讯作者：
Pourquie, Olivier
中科院分区：
文献类型：
--
作者：
Chal, Jerome;Al Tanoury, Ziad;Pourquie, Olivier
由于缺乏相关的细胞模型，也缺乏用于生物医学研究的可靠的肌肉前体细胞来源，因此在寻找肌肉疾病的治疗方法方面进展缓慢。在这里，我们报道了一种优化的无血清分化方案，可以在体外高效地从人多能干细胞(HPSC)中产生毫米长的横纹肌肉纤维和卫星样细胞。通过模拟导致胚胎肌肉形成的关键信号事件，特别是Wnt和骨形态发生蛋白(BMP)途径信号的双重调节，这种定向分化协议避免了遗传修改或细胞分选的要求。有hPSC培养经验的人员可以在1个月内在体外实现强健的肌肉生成。分化培养物可进行继代培养，以产生大量的肌源性祖细胞，可用于多种下游应用。除了对肌发生的研究外，这种分化方法还为发展相关的肌营养不良症体外模型和药物筛选策略提供了一个有吸引力的平台，并为组织工程和细胞治疗方法提供了细胞来源。
Progress toward finding a cure for muscle diseases has been slow because of the absence of relevant cellular models and the lack of a reliable source of muscle progenitors for biomedical investigation. Here we report an optimized serum-free differentiation protocol to efficiently produce striated, millimeter-long muscle fibers together with satellite-like cells from human pluripotent stem cells (hPSC) in vitro. By mimicking key signaling events leading to muscle formation in the embryo, in particular the dual modulation of Wnt and bone morphogenetic protein (BMP) pathway signaling, this directed differentiation protocol avoids the requirement for genetic modifications or cell sorting. Robust myogenesis can be achieved in vitro within 1 month by personnel experienced in hPSC culture. The differentiating culture can be subcultured to produce large amounts of myogenic progenitors amenable to numerous downstream applications. Beyond the study of myogenesis, this differentiation method offers an attractive platform for the development of relevant in vitro models of muscle dystrophies and drug screening strategies, as well as providing a source of cells for tissue engineering and cell therapy approaches.