胚胎干细胞来源的内皮细胞构建内皮化血管支架

目的探索胚胎干细胞定向分化为血管内皮细胞,并用于构建内皮化血管支架,为临床心血管疾病治疗提供实验基础。方法采用改良后单层分化体系,血管内皮生长因子（VEGF）作为独立诱导因子定向诱导胚胎干细胞分化为血管内皮细胞,用RT-PCR法检测分化时程各时间点内皮特异性标志的表达,用免疫荧光法检测分化体系中的内皮细胞蛋白标志。将分化成熟的内皮细胞种植于支架表面,构建内皮化血管支架。结果胚胎干细胞全能性分子标志Oct3／4,早期内皮标志Flk-1,Tie-2,PECAM-1,以及成熟内皮标志VE-cadherin在胚胎干细胞分化体系中呈现一定的转录时序性;免疫荧光结果显示分化细胞Flk-1,VE-cadheiin阳性,血管支架被表达VE-cadherin的内皮细胞覆盖。结论我们改良了胚胎干细胞分化为血管内皮细胞的方法,并用于探索内皮化血管支架的构建。

Seeding of vascular stent with endothelial cells derived from embryonic stem cells

Purpose In order to construct an endothelialized vascular stent, we differentiated embryonic stem cells (ES) into endothelial cells and seeded them onto the vascular stents, which would be a new therapeutic approach for cardiovascular disease. Methods Embryonic stem cells were differentiated into endothelial cells with improved monolayer differentiation in which vascular endothelial growth factor (VEGF) acted as the only induction factor. ES and endothelial-specific molecular markers were detected by RT-PCR at different time-points. The ES derived-endothelial cells were seeded onto the vascular stents which were verified by immunofluorescence. Results ES-derived cells expressed a full range of cell lineage-specific markers: Oct3/4, Flk-1, Tie-2, platelet endothelial cell adhesion molecule-1 (PECAM-1), vascular endothelial (VE)-cadherin by RT-PCR and immunofluorescence. The endothelialization stent was shown on the stent by VE-cadherin immunofluorescence. Conclusions Stent endothelialization was fulfilled by seeding endothelial cells derived from embryonic stem cells onto vascular stent.