血管内皮构成血管的内层,和功能障碍和损伤的内皮会导致心血管疾病以及其他疾病,包括中风,肿瘤生长,和慢性肾衰竭.产生有效的来源来替代受损的内皮细胞(ECs)可能具有显著的临床影响,和体细胞来源,如外周血或脐带血不能可靠地提供足够的内皮细胞祖细胞进行多种治疗。多能干细胞是可靠的EC供应的有希望的来源,具有恢复组织功能和治疗血管疾病的潜力。我们已经开发了将诱导多能干细胞(iPSC)在多个iPSC细胞系中有效且稳健地分化为高纯度的非组织特异性泛血管EC(iEC)的方法。这些iEC具有典型的内皮细胞标记物,并通过摄取Dil荧光染料标记的乙酰化低密度脂蛋白(Dil-Ac-LDL)和管形成表现出内皮细胞功能的量度。使用蛋白质组学分析,我们发现,iECs与已建立的人脐静脉ECs(HUVECs)相比,在蛋白组学上更相似。翻译后修饰(PTM)在HUVEC和IEC之间最共有,并确定了增加iECs与HUVECs蛋白质组相似性的潜在靶标。在这里,我们展示了一种将iPSCs分化为功能性ECs的高效稳健方法,并首次提供了iECs的全面蛋白质表达谱,这表明它们与广泛使用的永生化HUVEC相似,允许对EC发展进行进一步的机械研究,信令,以及未来再生应用的新陈代谢。NEW&NOTEWORTHY我们已经开发了将多个iPSC系的诱导多能干细胞(iPSC)分化为非组织特异性全血管EC(iEC)的方法,并证明了这些细胞与广泛使用的内皮细胞系(HUVEC)的蛋白质组学相似性。我们还鉴定了用于增加iECs与HUVECs的蛋白质组相似性的翻译后修饰和靶标。在未来,iEC可用于研究EC的发展,信令,以及未来再生应用的新陈代谢。
The vascular endothelium constitutes the inner lining of the blood vessel, and malfunction and injuries of the endothelium can cause cardiovascular diseases as well as other diseases including stroke, tumor growth, and chronic kidney failure. Generation of effective sources to replace injured endothelial cells (ECs) could have significant clinical impact, and somatic cell sources like peripheral or cord blood cannot credibly supply enough endothelial cell progenitors for multitude of treatments. Pluripotent stem cells are a promising source for a reliable EC supply, which have the potential to restore tissue function and treat vascular diseases. We have developed methods to differentiate induced pluripotent stem cells (iPSCs) efficiently and robustly across multiple iPSC lines into nontissue-specific pan vascular ECs (iECs) with high purity. These iECs present with canonical endothelial cell markers and exhibit measures of endothelial cell functionality with the uptake of Dil fluorescent dye-labeled acetylated low-density lipoprotein (Dil-Ac-LDL) and tube formation. Using proteomic analysis, we revealed that the iECs are more proteomically similar to established human umbilical vein ECs (HUVECs) than to iPSCs. Posttranslational modifications (PTMs) were most shared between HUVECs and iECs, and potential targets for increasing the proteomic similarity of iECs to HUVECs were identified. Here we demonstrate an efficient robust method to differentiate iPSCs into functional ECs, and for the first time provide a comprehensive protein expression profile of iECs, which indicates their similarities with a widely used immortalized HUVECs, allowing for further mechanistic studies of EC development, signaling, and metabolism for future regenerative applications.NEW & NOTEWORTHY We have developed methods to differentiate induced pluripotent stem cells (iPSCs) across multiple iPSC lines into nontissue-specific pan vascular ECs (iECs) and demonstrated the proteomic similarity of these cells to a widely used endothelial cell line (HUVECs). We also identified posttranslational modifications and targets for increasing the proteomic similarity of iECs to HUVECs. In the future, iECs can be used to study EC development, signaling, and metabolism for future regenerative applications.