贵州医科大学学报

2006, (06) 499-501

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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

王志华;何志旭;汪浩文;米蔷;
WANG Zhi-hua* , HE Zhi-xu, WANG Hao-wen, Mi Qiang (1.Department of Pediatrics, Guiyang Medical College, Guiyang 550004, China)

摘要(Abstract):

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。
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% CO_2 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.

关键词(KeyWords): 内皮,血管;脐静脉;细胞培养
endothelium,vascular; umbilical veins; cell culture

Abstract:

Keywords:

基金项目(Foundation): 贵州省省长基金资助项目[黔科教办(200614)]

作者(Author): 王志华;何志旭;汪浩文;米蔷;
WANG Zhi-hua* , HE Zhi-xu, WANG Hao-wen, Mi Qiang (1.Department of Pediatrics, Guiyang Medical College, Guiyang 550004, China)

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DOI: 10.19367/j.cnki.1000-2707.2006.06.004

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