WNT3A promotes myogenesis of human embryonic stem cells and enhances in vivo engraftment.
WNT3A promotes myogenesis of human embryonic stem cells and enhances in vivo engraftment.
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DOI：
10.1038/srep05916
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发表时间：
2014-08-01
影响因子：
4.6
通讯作者：
Varghese S
中科院分区：
文献类型：
--
作者：
Hwang Y;Suk S;Shih YR;Seo T;Du B;Xie Y;Li Z;Varghese S
人胚胎干细胞（human embryonic stem cells，hESC）分化为骨骼肌细胞的能力是其作为改善骨骼肌损伤的细胞来源的重要标准。然而，将hESC分化为骨骼肌细胞仍然是一个挑战，通常需要引入转基因。在这里，我们描述了使用WNT 3A蛋白来促进hESC衍生的细胞的体外成肌承诺及其随后的体内功能。我们的研究结果表明，培养基中WNT 3A的存在显著促进表达中胚层标记物血小板衍生生长因子受体-α（PDGFRA）的hESC衍生祖细胞的成肌定型，这从成肌标记物（包括DES、MYOG、MYH 1和MF 20）的表达中可以看出。将这些定向细胞移植到NOD/SCID小鼠心脏毒素损伤的骨骼肌中，揭示了供体细胞的存活和植入。这些细胞有助于受损肌纤维和卫星细胞室的再生。代替用于治疗骨骼肌缺陷的有限细胞来源，hESC衍生的PDGFRA+细胞表现出显著的体外扩增，同时保持其肌原性潜力。本研究中描述的结果提供了具有体内植入潜力的肌源性祖细胞可以在没有遗传操作的情况下从hESC衍生的原理证明。
The ability of human embryonic stem cells (hESCs) to differentiate into skeletal muscle cells is an important criterion in using them as a cell source to ameliorate skeletal muscle impairments. However, differentiation of hESCs into skeletal muscle cells still remains a challenge, often requiring introduction of transgenes. Here, we describe the use of WNT3A protein to promote in vitro myogenic commitment of hESC-derived cells and their subsequent in vivo function. Our findings show that the presence of WNT3A in culture medium significantly promotes myogenic commitment of hESC-derived progenitors expressing a mesodermal marker, platelet-derived growth factor receptor-α (PDGFRA), as evident from the expression of myogenic markers, including DES, MYOG, MYH1, and MF20. In vivo transplantation of these committed cells into cardiotoxin-injured skeletal muscles of NOD/SCID mice reveals survival and engraftment of the donor cells. The cells contributed to the regeneration of damaged muscle fibers and the satellite cell compartment. In lieu of the limited cell source for treating skeletal muscle defects, the hESC-derived PDGFRA+ cells exhibit significant in vitro expansion while maintaining their myogenic potential. The results described in this study provide a proof-of-principle that myogenic progenitor cells with in vivo engraftment potential can be derived from hESCs without genetic manipulation.