人脐静脉血管内皮细胞的培养和鉴定

The Culture and Identification of Human Umbilical Vein Endothelial Cells

目的：建立血管内皮细胞体外培养的模型，为研究内皮细胞提供重要的实验方法。方法：采用0．1％Ⅰ型胶原酶消化灌注脐静脉，在37℃水浴箱内孵育18～20rain，离心后向沉淀内加入L-DMEM培养液（含10mg／L内皮细胞生长因子），吹打成单细胞悬液，放入预铺1．5％明胶的培养瓶内，在CO2培养箱中培养，48h后换液1次，以后每2—3d换液1次，每天在倒置相差显微镜下观察细胞，免疫组织化学法进行Ⅷ因子相关抗原鉴定。结果：经胶原酶消化法获得的细胞经过镜下观察和Ⅷ因子相关抗原鉴定证明是脐静脉内皮细胞。结论：用胶原酶消化法是获得血管内皮细胞的一种好方法，获得的细胞数量多、成活率高。

Objective： To establish an experimental model of human vascular endothelial cell （EC） in vitro with human umbilical vein ECs. Methods： ECs were obtained by perfusing type I collagenase （0.1%） and to digest umbilical vein. The ECs collected by centrifuge were cultured in low-Dulbecco＇s modified Eagle＇s medium with 10% FBS plus penicillin, streptomycin, vascular endothelial growth factor and kept at 37℃ in a 5% CO2 air atmosphere. The cultured ECs were observed under inverted microscope for their morphologie characteristics and were identified by immunoehemical method with factor Ⅷ antigen. Results： The cultured cells were identified as umbilical vein ECs. Conclusion： The digestion using collagenase is a good method for collection of human umbilical vein ECs.