目的探讨从大鼠外周血中获取内皮前细胞的方法,为缺血性心脏病细胞移植治疗提供新的移植材料。方法首先应用密度梯度离心法分取大鼠外周血中的单个核细胞,然后以IMEM、胎牛血清、马血清、VEGF、bFGF等混合而成的培养基,进行体外培养。最后将此方法收获的细胞经一系列免疫荧光染色,并检测其结合UEA-1、摄取acLDL的能力。结果细胞CD31、CD34、Flk-1和vWF免疫荧光染色阳性,并能结合UEA-1、摄取acLDL。同时结合细胞的形态学变化特点,证实得到的细胞为内皮前细胞。结论通过一定的体外分离、培养途径,可以从大鼠外周血获得较为纯化的内皮前细胞。
Objective To explore the procedures of isolation, cultivation and purification of endothelial progenitor cells (EPC) from peripheral bloods. Methods Ten mL of peripheral blood was obtained from the femoral artery of each Sprague Dawley (SD) rat. Mononuclear cells were isolated by Ficoll density gradient centrifugation cell and cultured in a special medium. Results Ceils harvested by this procedure were immunohistologically proved EPC. Conclusion Relatively purified EPC can be obtained by certain procedure of isolation and culture.