人脐带血内皮祖细胞的分离、培养及鉴定  

Isolation,culture and identification of endothelial progenitor cells in human umbilical cord blood

作　　者：冯小敏 陈翔[2] FENG Xiao-min(Department ofpaediatrics,the first affiliated hospital of Wannan Medical College,Yi Ji Shan Hospital,Wuhu,Anhui,241000,China)

机构地区：[1]皖南医学院第一附属医院弋矶山医院儿科,安徽芜湖241000 [2]温州医科大学附属第二医院康复科,浙江温州325027

出　　处：《齐齐哈尔医学院学报》2020年第4期397-400,共4页Journal of Qiqihar Medical University

摘　　要：目的从健康足月新生儿脐带血中分离内皮祖细胞进行培养鉴定,为后期研究提供细胞来源。方法采用密度梯度离心法获取单个核细胞,接种在鼠尾Ⅰ型胶原包被的培养板中,用内皮细胞培养液EGM-2(endothelial cell growth medium-2)培养,观察脐带血来源的内皮祖细胞(Endothelial progenitor cells,EPCs)增殖生长情况,通过观察细胞形态、双荧光染色法、免疫荧光细胞化学染色法及流式细胞仪等技术对培养的内皮祖细胞进行鉴定。结果人脐带血可分离获得内皮祖细胞;脐带血内皮祖细胞在培养10~21 d出现,表现为贴壁和典型的"铺路石样"形态。采用双荧光染色法、免疫荧光细胞化学染色法及流式细胞仪可鉴定EPCs。结论采用密度梯度离心法可获得较好的人脐带血来源内皮祖细胞。Objective To isolate endothelial progenitor cells(EPCs)from the cord blood of newly-born healthy full-term babies for culture and identification and to provide evidences and cells for future studies.Methods Density gradient centrifugation was adopted to obtain mononuclear cells which were then inoculated to the culture plates coated with rat tail collagen-I and cultured in the endothelial cell nutrient solution of EGM-2(endothelial cell growth medium-2).The proliferation and growth of the cord blood-derived EPCs were observed and identified through cellular morphology observation,double fluorescence staining,immunofluorescence cytochemistry staining,flow cytometry,etc.Results EPCs could be separated and obtained from human cord blood;cord blood EPCs were generated after being cultured for 10 to 21 days,presented as adherent cells and the typical"cobblestone"morphology;and double fluorescence staining,immunofluorescence cytochemistry staining and flow cytometry could be used to identify EPCs.Conclusions Preferable human cord blood-derived EPCs could be obtained by using the method of density gradient centrifugation.

关 键 词：内皮祖细胞 脐带血 分离 培养 鉴定 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]