Robust differentiation of human pluripotent stem cells into endothelial cells via temporal modulation of ETV2 with modified mRNA
Robust differentiation of human pluripotent stem cells into endothelial cells via temporal modulation of ETV2 with modified mRNA
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DOI：
10.1126/sciadv.aba7606
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发表时间：
2020-07-01
期刊：
影响因子：
13.6
通讯作者：
Melero-Martin, Juan M.
中科院分区：
文献类型：
--
作者：
Wang, Kai;Lin, Ruei-Zeng;Melero-Martin, Juan M.
人诱导多能干细胞（h-iPSC）衍生的内皮细胞（h-iEC）已成为再生医学中的有价值的工具。然而，目前的分化方案仍然效率低下且缺乏可靠性。在这里，我们描述了一种快速、一致和高效生成h-iEC的方法。该方案需要在分化的中间中胚层阶段递送编码转录因子ETV 2的修饰的mRNA。该方法以极高的效率可重复地将13种不同的h-iPSC系分化成h-iEC。相比之下，依赖于内源性ETV 2的标准分化方法是低效的并且显著不一致。我们的h-iEC在许多方面都具有功能，包括在体内形成灌注血管网络的能力。ETV 2的及时激活是至关重要的，并且绕过中胚层阶段产生了具有降低的扩张潜力并且不能形成功能性血管的推定的h-iEC。我们的方案具有广泛的应用，并且可以可靠地为血管治疗提供无限数量的h-iEC。
Human induced pluripotent stem cell (h-iPSC)-derived endothelial cells (h-iECs) have become a valuable tool in regenerative medicine. However, current differentiation protocols remain inefficient and lack reliability. Here, we describe a method for rapid, consistent, and highly efficient generation of h-iECs. The protocol entails the delivery of modified mRNA encoding the transcription factor ETV2 at the intermediate mesodermal stage of differentiation. This approach reproducibly differentiated 13 diverse h-iPSC lines into h-iECs with exceedingly high efficiency. In contrast, standard differentiation methods that relied on endogenous ETV2 were inefficient and notably inconsistent. Our h-iECs were functionally competent in many respects, including the ability to form perfused vascular networks in vivo. Timely activation of ETV2 was critical, and bypassing the mesodermal stage produced putative h-iECs with reduced expansion potential and inability to form functional vessels. Our protocol has broad applications and could reliably provide an unlimited number of h-iECs for vascular therapies.