文章摘要
付 伟,李慧敏,苏建华,缪 珩,鲁一兵.人脐静脉内皮细胞的原代培养及鉴定[J].南京医科大学学报,2009,29(1):89~91
人脐静脉内皮细胞的原代培养及鉴定
Culture and identification of human umbilical vein endothelial cells
投稿时间:2008-07-10  
DOI:10.7655
中文关键词: 人脐静脉内皮细胞  培养  鉴定  CD34
英文关键词: human umbilical endothelial cells(HUVECs)  culture  identification  CD34
基金项目:江苏省卫生厅面上项目(H200514)
作者单位
付 伟 南京医科大学第二附属医院内分泌科,江苏 南京 210011 
李慧敏  
苏建华  
缪 珩  
鲁一兵  
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中文摘要:
      目的:体外进行人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)原代培养及鉴定?方法:用0.1%Ⅱ型胶原酶消化法收集HUVECs,加入含20%胎牛血清的M199培养基在37℃?5%CO2孵箱中培养,用Tryple express进行消化传代培养?采用细胞形态学观察?流式细胞仪测定CD34鉴定细胞?结果:原代培养的HUVECs在24 h完全贴壁,在3~5天融合成片,呈现单层铺路石样改变?流式细胞仪测定内皮细胞纯度为75.52%?结论:Ⅱ胶原酶及Tryple express的灵活使用可获得大量连续传代培养的HUVECs?
英文摘要:
      Objective:To culture and identify endothelial cells from human umbilical vein endothelial cells(HUVECs) in vitro. Methods:HUVECs were obtained using 0.1% collagenase typeⅡ by the technique of irrigative digestion, and were cultured with M199 which contained fetal calf serum in the carbon dioxide incubator,and then were passaged using tryple express digestion. The culture cells were identified by morphology and flow cytometry detecting CD34. Results:HUVECs could adhere to the plate completely after 24 hours and got a monolayer 3~5 days later. The culture cells were HUVECs confirmed by morphology and flow cytometry. Conclusion:Using collagenase typeⅡand tryple express could obtain a large number of HUVECs in continuous passage culture.
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