Efficient generation of endothelial cells from human pluripotent stem cells and characterization of their functional properties.
Efficient generation of endothelial cells from human pluripotent stem cells and characterization of their functional properties.
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DOI：
10.1002/jbm.a.35607
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发表时间：
2016-03
期刊：
影响因子：
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通讯作者：
Wei Song;D. Kaufman;W. Shen
中科院分区：
文献类型：
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作者：
Wei Song;D. Kaufman;W. Shen
虽然内皮细胞（ECs）已经从人多能干细胞（hPSCs）中衍生出来，但大规模生成hPSC-ECs仍然具有挑战性，其功能也没有很好地表征。在这里，我们报告了一种简单有效的三步法，可以在第16天和第34天分别从每个人胚胎干细胞（hESC）输入中产生大约98和9500个内皮细胞。研究了tgf β抑制分子SB431542存在和不存在情况下获得的hESC-ECs的功能特性，并与人脐静脉内皮细胞（HUVECs）的功能特性进行了比较。SB431542 + hESC-ECs、SB431542- hESC-ECs和HUVECs形成的融合单层对10000 Da葡聚糖的通透性相似，但这些细胞在三维纤维蛋白凝胶中形成管状结构的能力存在显著差异。无论培养基中是否存在VEGF和bFGF， SB431542 + hesc - ec在形成管状结构方面最有效；弱效SB431542- hESC-ECs和HUVECs对VEGF和bFGF的反应不同，bFGF显著增强了HUVECs形成管状结构的能力，但对SB431542- hESC-ECs影响不大。本研究为大规模生产hPSC-EC提供了一种有效的方法，并表明在技术开发中使用之前，需要对不同条件下获得的hPSC-EC的表型和功能进行彻底的检查。©2015 Wiley期刊公司[J] .生物医学工程学报，2016,35(4):678-687。
Although endothelial cells (ECs) have been derived from human pluripotent stem cells (hPSCs), large-scale generation of hPSC-ECs remains challenging and their functions are not well characterized. Here we report a simple and efficient three-stage method that allows generation of approximately 98 and 9500 ECs on day 16 and day 34, respectively, from each human embryonic stem cell (hESC) input. The functional properties of hESC-ECs derived in the presence and absence of a TGFβ-inhibitory molecule SB431542 were characterized and compared with those of human umbilical vein endothelial cells (HUVECs). Confluent monolayers formed by SB431542 + hESC-ECs, SB431542- hESC-ECs, and HUVECs showed similar permeability to 10,000 Da dextran, but these cells exhibited striking differences in forming tube-like structures in 3D fibrin gels. The SB431542 + hESC-ECs were most potent in forming tube-like structures regardless of whether VEGF and bFGF were present in the medium; less potent SB431542- hESC-ECs and HUVECs responded differently to VEGF and bFGF, which significantly enhanced the ability of HUVECs to form tube-like structures but had little impact on SB431542- hESC-ECs. This study offers an efficient approach to large-scale hPSC-EC production and suggests that the phenotypes and functions of hPSC-ECs derived under different conditions need to be thoroughly examined before their use in technology development. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 678-687, 2016.