利用内皮祖细胞制备组织工程血管的实验研究

Construction of tissue engineering vessels from endothelial progenitor cells

目的:观察内皮祖细胞(endothelial progen itor cells,EPCs)在去细胞血管段上的再内皮化效果,探讨其组织工程学应用前景。方法:采用贴壁选择法培养人外周血EPCs,VEGF扩增分化,流式细胞仪分析培养细胞CD34、VE-Cadherin的表达,D iI-ac-LDL吞噬试验、Ⅷ因子相关抗原免疫组化及细胞形态观察证实培养细胞的内皮属性。采用球囊损伤法制备去内膜兔腹主动脉段,去内皮兔腹主动脉段体外培养3周使之去细胞,与人外周血EPCs共孵育制备再内皮化异种去细胞血管段。结果:体外成功培养出人外周血内皮祖细胞,EPCs与去细胞血管段共培育使之再内皮化,形成新内膜。结论:体外培养EPCs可制备再内皮化的异种去细胞血管段,提供了一种制备组织工程血管的新方法。

Abstract： AIM： To investigate the reendothelialization of decellular vessels via endothelial progenitor cells （EPCs） ,and study its feasibility for tissue engineering application. METHODS：Human peripheral blood-derived endothelial progenitor cells （PB EPCs） were cultured in vitro through adhesion selection and differentiated by VEGF. To confirm that the cultured cells were EPCs, fluorescence -activated cell sorting （FACS） analysis was performed to detect the expression of surface marker CD34 and VE-Cadherino An EC lineage was confirmed by immunocytochemistry of Ⅷ factor relative antigen, the uptake of Dilac-LDL and fluorescent microscopic observation. To study the feasibility of in vitro reendothelialization via EPCs, rabbit abdominal aorta segments obtained by balloon catheter injury were decellularized after a 3 week in vitro culture and were co-incubated with human PB EPCs for a week to make themselves reendothelialized. RESULTS： Human PB EPCs were successfully cultured in vitro. And coincubating decellular rabbit abdomial aorta segments with cultured human PB EPCs contributed to the reendothelialization of abdomial aorta segments. CONCLUSION： Cultured human PB EPCs can be used to construct heterogenous reendothelialized decellular vessel segments, which will provide a novel way to construct tissue engineering vessels.