Medical Faculty, Center for Physiology and Pathophysiology, Institute for Neurophysiology, University of Cologne, Robert Koch Str. 39, 50931, Cologne, Germany.
Biology Department, Faculty of Science, Soran University, Kurdistan Region, Soran, Iraq.
Stem Cell Res Ther. 2022 Jun 11;13(1):251. doi: 10.1186/s13287-022-02924-x.
Endothelial cells (ECs) form the inner lining of all blood vessels of the body play important roles in vascular tone regulation, hormone secretion, anticoagulation, regulation of blood cell adhesion and immune cell extravasation. Limitless ECs sources are required to further in vitro investigations of ECs' physiology and pathophysiology as well as for tissue engineering approaches. Ideally, the differentiation protocol avoids animal-derived components such as fetal serum and yields ECs at efficiencies that make further sorting obsolete for most applications.
Human induced pluripotent stem cells (hiPSCs) are cultured under serum-free conditions and induced into mesodermal progenitor cells via stimulation of Wnt signaling for 24 h. Mesodermal progenitor cells are further differentiated into ECs by utilizing a combination of human vascular endothelial growth factor A165 (VEGF), basic fibroblast growth factor (bFGF), 8-Bromoadenosine 3',5'-cyclic monophosphate sodium salt monohydrate (8Bro) and melatonin (Mel) for 48 h.
This combination generates hiPSC derived ECs (hiPSC-ECs) at a fraction of 90.9 ± 1.5% and is easily transferable from the two-dimensional (2D) monolayer into three-dimensional (3D) scalable bioreactor suspension cultures. hiPSC-ECs are positive for CD31, VE-Cadherin, von Willebrand factor and CD34. Furthermore, the majority of hiPSC-ECs express the vascular endothelial marker CD184 (CXCR4).
The differentiation method presented here generates hiPSC-ECs in only 6 days, without addition of animal sera and at high efficiency, hence providing a scalable source of hiPSC-ECs.
内皮细胞（ECs）构成了体内所有血管的内层，在血管张力调节、激素分泌、抗凝、调节血细胞黏附和免疫细胞渗出等方面发挥着重要作用。为了进一步研究 ECs 的生理学和病理生理学，以及组织工程方法，需要有无尽的 ECs 来源。理想情况下，分化方案避免使用动物源性成分，如胎牛血清，并以效率产生 ECs，使大多数应用程序不再需要进一步的分选。
人诱导多能干细胞（hiPSCs）在无血清条件下培养，并通过刺激 Wnt 信号 24 小时诱导成中胚层祖细胞。通过利用人血管内皮生长因子 A165（VEGF）、碱性成纤维细胞生长因子（bFGF）、8-溴腺苷 3'，5'-环单磷酸钠盐一水合物（8Bro）和褪黑素（Mel）的组合，将中胚层祖细胞进一步分化为 ECs，持续 48 小时。
该组合以 90.9±1.5%的分数生成 hiPSC 衍生的 ECs（hiPSC-ECs），并且很容易从二维（2D）单层转移到三维（3D）可扩展的生物反应器悬浮培养物中。hiPSC-ECs 对 CD31、VE-钙粘蛋白、血管性血友病因子和 CD34 呈阳性。此外，大多数 hiPSC-ECs 表达血管内皮标志物 CD184（CXCR4）。
这里提出的分化方法仅需 6 天即可生成 hiPSC-ECs，无需添加动物血清且效率高，因此提供了一种可扩展的 hiPSC-ECs 来源。
