Enhanced Development of Skeletal Myotubes from Porcine Induced Pluripotent Stem Cells.
Enhanced Development of Skeletal Myotubes from Porcine Induced Pluripotent Stem Cells.
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DOI：
10.1038/srep41833
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发表时间：
2017-02-06
影响因子：
4.6
通讯作者：
Roberts RM
中科院分区：
文献类型：
--
作者：
Genovese NJ;Domeier TL;Telugu BP;Roberts RM
猪除了在农业上的重要性外，还被认为是生物医学研究中有价值的模型。在这里，我们描述了一种用于在体外从猪诱导多能干细胞（piPSC）有效地产生骨骼肌的方法，从而为从再生生物学到肉的离体培养的应用提供了通用平台。GSK 3B抑制剂CHIR 99021用于抑制细胞凋亡，引发WNT信号传导事件并沿着中胚层谱系驱动幼稚型piPSC，并且与DNA甲基化抑制剂5-氮杂胞苷组合，以激活早期骨骼肌转录程序。然后通过激活异位表达的MYOD 1诱导终末分化。肌管，其特征在于肌原纤维的发展和自发和刺激引起的兴奋-收缩偶联周期出现在11天内。通过均匀的NCAM 1和肌球蛋白重链表达证实了有效的谱系特异性分化。这些结果提供了一种产生骨骼肌的方法，该方法可能适用于其他多能细胞系和产生其他形式的肌肉。
The pig is recognized as a valuable model in biomedical research in addition to its agricultural importance. Here we describe a means for generating skeletal muscle efficiently from porcine induced pluripotent stem cells (piPSC) in vitro thereby providing a versatile platform for applications ranging from regenerative biology to the ex vivo cultivation of meat. The GSK3B inhibitor, CHIR99021 was employed to suppress apoptosis, elicit WNT signaling events and drive naïve-type piPSC along the mesoderm lineage, and, in combination with the DNA methylation inhibitor 5-aza-cytidine, to activate an early skeletal muscle transcription program. Terminal differentiation was then induced by activation of an ectopically expressed MYOD1. Myotubes, characterized by myofibril development and both spontaneous and stimuli-elicited excitation-contraction coupling cycles appeared within 11 days. Efficient lineage-specific differentiation was confirmed by uniform NCAM1 and myosin heavy chain expression. These results provide an approach for generating skeletal muscle that is potentially applicable to other pluripotent cell lines and to generating other forms of muscle.