赵清,魏盟,赵炳辉,刘一文,何怡青

• 1:上海交通大学附属第六人民医院心内科
• 2:上海交通大学附属第六人民医院放射科
• 3:上海交通大学附属第六人民医院中心实验室

摘要(Abstract)：

目的体外制备兔内皮祖细胞(EPC)种植支架,并评价其可行性。方法采用密度梯度离心法分离兔外周血单个核细胞,接种于人纤维连接蛋白包被的培养板上,予含内皮细胞生长添加剂30μg/mL的M199培养基培养。培养2周后进行免疫荧光染色和免疫细胞化学法鉴定EPC,检测其增殖、迁移、黏附和分泌血管内皮生长因子(VEGF)、粒细胞集落刺激因子(G-CSF)和一氧化氮(NO)的能力。培养的EPC消化后,将其浇注到有人纤维连接蛋白包被和无包被的裸支架上,6 d后行扫描电镜和荧光显微镜检查,观察其表面细胞的生长情况。结果单个核细胞培养14 d后,出现大量梭形细胞,免疫荧光染色显示细胞可同时摄取Dil标记的乙酰化低密度脂蛋白和结合FITC标记的凝集素者,表明为正在分化的EPC,同时证实其表达内皮细胞的特异标志vWF。培养2周后的EPC数量呈指数增加;EPC的黏附率为96.7%±3.2%;迁移为15±4细胞/×400视野;可分泌VEGF、G-CSF和NO。与无包被的裸支架组相比,有纤维连接蛋白包被的支架组,EPC黏附多(27.80±4.26细胞/×400视野比6.10±3.07细胞/×400视野,P<0.05),内皮化完全。结论EPC可向内皮细胞方向分化,并且具有高增殖、高迁移和高黏附的能力。采用EPC在体外制备细胞种植支架是可行的,而纤维连接蛋白可加速其黏附。

关键词(KeyWords)： 兔;干细胞;支架

Abstract:

Keywords:

基金项目(Foundation): 上海市科委课题(编号05JC14031)

作者(Author): 赵清,魏盟,赵炳辉,刘一文,何怡青

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