摘要
目的探讨人脐静脉内皮细胞(Human umbilical vein endothelial cell,HUVEC)体外原代培养及功能检测的可靠方法。方法采用改良的胶原酶灌注消化法分离HUVEC,用免疫荧光法鉴定细胞纯度;用血清+DMSO冻存HUVEC,用Edu法检测细胞增殖能力,Matrigel体外成管实验检测血管内皮细胞功能。结果培养得到高纯度(99%)、高数量(1×106~3×106)、高质量(增殖与成管能力均良好)的原代细胞。结论建立了分离、培养、保存HUVEC及对其功能检测的可靠方法,为血管新生研究提供了理想的细胞模型和实验思路。
Objective To explore a reliable method for primary culture,identification and reliable function experiment of human umbilical vein endothelial cells. Methods HUVECs were separately cultured by improved collagenase perfusion digestion method. Cell purity was identify by immunofluorescence while HUVECs were stored using serum and DMSO frozen. Edu mixing method were used to detect cell proliferation and Matrigel tube formation assay were used to test cell fuction. Results The amounts of the endothelial cells isolated from HUVEC were( 1 ~ 3) × 106cells. More than 99% cells were positive identified by factor VIII and CD31. Over90% of the cells were viable from revivification. Conclusion We established a reliable method of cultivation,storage,revivification,identification and function test of HUVECs in vitro.
出处
《宁夏医科大学学报》
2013年第11期1204-1207,1188,共5页
Journal of Ningxia Medical University
基金
国家自然科学基金(81360094
81260110)
宁夏教育厅重点项目(宁教高[2011]263)
关键词
人脐静脉内皮细胞
原代培养
鉴定
血管功能实验
human umbilical vein endothelial
cell primary culture
identification
function experiment
