Department of Cardiology, Shanghai Sixth People's Hospital, Shanghai Jiao Tong University, 200233, China.
Scand Cardiovasc J. 2010 Apr;44(2):76-81. doi: 10.1080/14017430903118165.
The aim of the study is to fabricate stents with rabbit outgrowth endothelial progenitor cells (OECs) to facilitate their endothelial function in vitro.
Rabbit OECs were isolated from peripheral blood and identified by immunofluorescence and flow cytometry. Cells proliferation and migration were measured by growth curve and modified Boyden chamber assay. Adhesion assay was performed by replating cells on fibronectin-coated dishes. VEGF, G-CSF and NO in supernatant were tested. OECs were poured on fibronectin-coated or uncoated stents. After six days, scanning electron microscopy (SEM) and inverted fluorescent microscopy observation were performed.
About three to four weeks after culture, OECs were characterized as adherent cells which were double positive for Dil-acLDL uptake and FITC-UEA-I binding, with high expression of CD34. They also showed high ability of proliferation, adhesion and migration properties. Compared with uncoated stents, more OECs migrated and adhered onto fibronectin-coated stents. OECs seeding onto the fibronectin coated stents could secret more cytokine and NO. Endothelialization of coated stents was visible both under the SEM and inverted fluorescent microscope.
OECs can differentiate to endothelial lineage and possess high ability of proliferation, migration and adhesion. It is feasible to fabricate OECs-seeded stents in vitro, while the stents coated with fibronectin facilitate this endothelialization process.
本研究旨在制备载有兔源性血管外膜祖细胞（OECs）的支架，以促进其体外内皮功能。
从外周血中分离兔源性 OECs，通过免疫荧光和流式细胞术进行鉴定。通过生长曲线和改良 Boyden 室测定法测量细胞增殖和迁移。通过将细胞铺在纤维连接蛋白包被的培养皿上进行黏附试验。检测上清液中的 VEGF、G-CSF 和 NO。将 OECs 浇铸在纤维连接蛋白包被或未包被的支架上。六天后，进行扫描电子显微镜（SEM）和倒置荧光显微镜观察。
培养约三到四周后，OECs 表现为贴壁细胞，对 Dil-acLDL 摄取和 FITC-UEA-I 结合呈双阳性，CD34 表达水平高。它们还表现出高增殖、黏附和迁移能力。与未包被的支架相比，更多的 OECs 迁移并黏附在纤维连接蛋白包被的支架上。种植在纤维连接蛋白涂层支架上的 OECs 可以分泌更多的细胞因子和 NO。SEM 和倒置荧光显微镜下均可观察到涂层支架的内皮化。
OECs 可分化为内皮谱系，具有高增殖、迁移和黏附能力。在体外制备载有 OECs 的支架是可行的，而纤维连接蛋白涂层支架有助于这一内皮化过程。
