CNC UC- Center for Neurosciences and Cell Biology, University of Coimbra, 3004-517, Coimbra, Portugal.
Faculty of Medicine, University of Coimbra, 3000-354, Coimbra, Portugal.
Sci Rep. 2019 Mar 7;9(1):3826. doi: 10.1038/s41598-019-40417-9.
The current work reports the functional characterization of human induced pluripotent stem cells (iPSCs)- arterial and venous-like endothelial cells (ECs), derived in chemically defined conditions, either in monoculture or seeded in a scaffold with mechanical properties similar to blood vessels. iPSC-derived arterial- and venous-like endothelial cells were obtained in two steps: differentiation of iPSCs into endothelial precursor cells (CD31/KDR/VE-Cad/EphB2/COUP-TF) followed by their differentiation into arterial and venous-like ECs using a high and low vascular endothelial growth factor (VEGF) concentration. Cells were characterized at gene, protein and functional levels. Functionally, both arterial and venous-like iPSC-derived ECs responded to vasoactive agonists such as thrombin and prostaglandin E2 (PGE), similar to somatic ECs; however, arterial-like iPSC-derived ECs produced higher nitric oxide (NO) and elongation to shear stress than venous-like iPSC-derived ECs. Both cells adhered, proliferated and prevented platelet activation when seeded in poly(caprolactone) scaffolds. Interestingly, both iPSC-derived ECs cultured in monoculture or in a scaffold showed a different inflammatory profile than somatic ECs. Although both somatic and iPSC-derived ECs responded to tumor necrosis factor-α (TNF-α) by an increase in the expression of intercellular adhesion molecule 1 (ICAM-1), only somatic ECs showed an upregulation in the expression of E-selectin or vascular cell adhesion molecule 1 (VCAM-1).
目前的工作报道了人类诱导多能干细胞（iPSCs）衍生的动脉和静脉样内皮细胞（ECs）的功能特征，这些细胞是在化学定义条件下培养的，无论是在单核培养中还是在与血管具有相似机械性能的支架中接种。iPSC 衍生的动脉和静脉样内皮细胞通过两步获得：首先将 iPSC 分化为内皮前体细胞（CD31/KDR/VE-Cad/EphB2/COUP-TF），然后使用高浓度和低浓度的血管内皮生长因子（VEGF）将其分化为动脉和静脉样 ECs。细胞在基因、蛋白质和功能水平上进行了表征。在功能上，两种类型的 iPSC 衍生的动脉和静脉样 ECs 对血管活性激动剂（如凝血酶和前列腺素 E2（PGE））都有反应，与体细胞 ECs 相似；然而，动脉样 iPSC 衍生的 ECs 产生的一氧化氮（NO）和对剪切应力的伸长比静脉样 iPSC 衍生的 ECs 更高。当接种在聚己内酯（PCL）支架中时，两种细胞都能粘附、增殖并防止血小板激活。有趣的是，在单核培养或支架中培养的两种 iPSC 衍生的 ECs 都显示出与体细胞 ECs 不同的炎症特征。尽管体细胞和 iPSC 衍生的 ECs 都通过增加细胞间黏附分子 1（ICAM-1）的表达对肿瘤坏死因子-α（TNF-α）做出反应，但只有体细胞 ECs 显示出 E-选择素或血管细胞黏附分子 1（VCAM-1）的表达上调。
