大鼠外周血晚期内皮前体细胞体外培养研究

目的体外培养大鼠外周血内皮前体细胞（EPCs），观察细胞克隆形态并进行鉴定。方法密度梯度离心法分离SD大鼠外周血单个核细胞，EGM-2培养基离体培养。免疫荧光染色鉴定细胞CD34、CD31、CD133、FLk-1、vwF细胞表面标志。Real—TimePCR检测细胞CD34、CD133、FLk-1、eNOSmRNA表达。结果细胞培养10天后可见“铺路石样细胞”和少数“紊乱生长细胞”，传代培养后经免疫荧光及Real—TimePCR鉴定“铺路石样细胞”符合晚期EPCs特点，紊乱生长细胞不表达相应标志。结论通过体外分离长时培养可从大鼠外周血获得晚期内皮前体细胞，具有内皮细胞的特征。

In vitro culture of late-outgrowth endothelial progenitor cells from rat peripheral blood

Objective To evaluate the characterization of endothelial progenitor cells（EPCs） from rat peripheral blood cultured in vitro. Methods 10ml peripheral blood was obtained from the abdominal aorta. Mononuclcar cells were isolated by using Ficoll density gradient centrifugation method, and cultured in EGM- 2 medium. Result Late-EPCs could be obtained after 10 days. EPCs were able to line up in the cord-like structure and form clusters. Cells showed CD34 、 CD133 、 CD31 、 FLk-1, vWF、 UEA-1/acLDL positive. , and high expressed CD34、CD133 、FLk-1、eNOS mRNA. Irregular cells couldnk express corresponding markers. Conclusion Late-outgrowth endothelial progenitor cells could be obtained from rat peripheral blood through long-term in vitro culture. The cells show the characterization of endothelial cells.