Directed Differentiation of Human Pluripotent Stem Cells toward Skeletal Myogenic Progenitors and Their Purification Using Surface Markers.
Directed Differentiation of Human Pluripotent Stem Cells toward Skeletal Myogenic Progenitors and Their Purification Using Surface Markers.
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DOI：
10.3390/cells10102746
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发表时间：
2021-10-14
期刊：
影响因子：
6
通讯作者：
Darabi R
中科院分区：
文献类型：
--
作者：
Xu N;Wu J;Ortiz-Vitali JL;Li Y;Darabi R
体细胞重编程为诱导多能干细胞（iPSC）的进展为体外疾病建模，基因校正和基于干细胞的再生医学提供了强大的框架。在骨骼肌病症的情况下，iPSC可用于产生骨骼肌祖细胞以研究疾病机制，或用于治疗肌肉病症的实施。我们最近开发了一种改进的定向分化方法，用于从hiPSC衍生骨骼肌源性祖细胞。该方法允许使用小分子在人多能干细胞（ESC/iPSC）中进行短期（2周）和有效的骨骼肌原性诱导（45-65%的细胞），以诱导中胚层和随后的肌瘤祖细胞，而不需要任何基因整合或修饰。在初始分化后，可以使用表面标记（CD 10 + CD 24 −）从不需要的细胞中纯化骨骼肌原性祖细胞。这些肌源性祖细胞已经使用体外基因表达/分化谱以及营养不良（mdx）和肌肉损伤（VML）啮齿动物模型中的体内植入研究进行了广泛表征，并且已经证明能够植入并形成成熟的肌纤维以及接种肌肉干细胞。目前的协议描述了这种方法的详细，分步指南，并概述了重要的实验细节和故障排除点，其应用于任何人类多能干细胞。
Advancements in reprogramming somatic cells into induced pluripotent stem cells (iPSCs) have provided a strong framework for in vitro disease modeling, gene correction and stem cell-based regenerative medicine. In cases of skeletal muscle disorders, iPSCs can be used for the generation of skeletal muscle progenitors to study disease mechanisms, or implementation for the treatment of muscle disorders. We have recently developed an improved directed differentiation method for the derivation of skeletal myogenic progenitors from hiPSCs. This method allows for a short-term (2 weeks) and efficient skeletal myogenic induction (45–65% of the cells) in human pluripotent stem cells (ESCs/iPSCs) using small molecules to induce mesoderm and subsequently myotomal progenitors, without the need for any gene integration or modification. After initial differentiation, skeletal myogenic progenitors can be purified from unwanted cells using surface markers (CD10+CD24−). These myogenic progenitors have been extensively characterized using in vitro gene expression/differentiation profiling as well as in vivo engraftment studies in dystrophic (mdx) and muscle injury (VML) rodent models and have been proven to be able to engraft and form mature myofibers as well as seeding muscle stem cells. The current protocol describes a detailed, step-by-step guide for this method and outlines important experimental details and troubleshooting points for its application in any human pluripotent stem cells.
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影响因子：
8.8
作者：
Xi H;Fujiwara W;Gonzalez K;Jan M;Liebscher S;Van Handel B;Schenke-Layland K;Pyle AD
通讯作者：
Pyle AD
影响因子：
4.6
作者：
Wu J;Hunt SD;Xue H;Liu Y;Darabi R
通讯作者：
Darabi R
影响因子：
--
作者：
Gu M
通讯作者：
Gu M
影响因子：
1.2
作者：
Wu, Jianbo;Hunt, Samuel D.;Darabi, Radbod
通讯作者：
Darabi, Radbod
DOI：
10.1038/nrd.2016.245
发表时间：
2017-03
期刊：
Nature reviews. Drug discovery
影响因子：
--
作者：
Shi Y;Inoue H;Wu JC;Yamanaka S
通讯作者：
Yamanaka S