【摘要】 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18~20min,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。更多还原

【Abstract】 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 morphologic characteristics and were identified by immunochemical 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.更多还原