人外周血内皮祖细胞的分离、培养、扩增和鉴定  

solation,culture,amplification and identification of human endothelial progenitor cells from peripheral blood

作　　者：李文志[1] 董勇[1] 辛毅[1] 

机构地区：[1]首都医科大学附属北京安贞医院整形美容和激光医学科

出　　处：《中国美容医学》2012年第8期1318-1320,共3页Chinese Journal of Aesthetic Medicine

摘　　要：目的：研究人外周血内皮祖细胞的分离、培养、扩增和鉴定方法。方法：采用密度梯度离心法分离人外周血单个核细胞，用EGM-2培养基在多聚赖氨酸包被的培养板中进行诱导培养，动态地观察贴壁细胞的生长状况，免疫组织化学技术检测培养细胞表面标志物的表达。结果：人外周血单个核细胞经体外培养至第2天，呈贴壁生长，细胞形态为大小相近的圆球体；第4天一些细胞开始出现尾状物，形态呈蝌蚪样改变，部分细胞聚集形成细胞簇；至第7天细胞形态变长，呈梭形改变，细胞间相互围绕呈环形，并有集落出现，免疫组织化学染色C12133、CD34、VEGFR-2和Vlll~子表达均呈阳性。结论：运用本实验方法可以成功实现人外周血内皮祖细胞的分离、培养和扩增。Objective To study the methods of isolation, culture, amplification and identification of human endothelial progenitor cells from peripheral blood. Methods The mononuclearcells （MNCs） were isolated from the human peripheral anticoagulant blood sample by Ficoll-density centrifugantion.The isolated cells were induced and cultured in the plates coated with polylysine,to observe the growing conditions of the adherent cells dynamically,immunohistochemical technology was used to detect the surface maker of the cultivated cells. Results MNCs showed adherent growth when cultured in the second day,the bodys of the cells like a ball and have the similar size,in the fourth day,tails were appeared in some cells and they looked like tadpoles,some cells gathered into cell clusters,in the seven days,the cells got longer and showed spindle- shaped changes, some of them gathered around with each other like a ring and a colony maybe occurred then,CD133,CD34, VEGFR-2 and VIII factor were all positively stained at cytoplasm in most cells. Conclusion This method can be well used to isotate,culture,amptificate and identificate of human endothelial progenitor cells from peripheral blood.

关 键 词：内皮祖细胞 外周血 细胞培养 

分 类 号：Q813.1[生物学—生物工程]