人脐静脉血内皮克隆形成细胞的分离培养与鉴定  

Isolate,culture and identification of endothelial colony-forming cells (ECFCs) from human umbilical cord blood in vitro

作　　者：周丽[1] 常青[2] 李自成[1] 

机构地区：[1]暨南大学附属第一医院心内科,广东广州510630 [2]暨南大学医学院组织学与胚胎学系,广东广州510632

出　　处：《中国医学工程》2013年第5期7-8,11,共3页China Medical Engineering

基　　金：广东省科技计划项目(2010-1096-136)

摘　　要：目的体外建立一种简便的人脐静脉血内皮克隆形成细胞(ECFCs)分离培养方法,并对其进行鉴定。方法无菌条件下收集脐静脉血,以密度梯度离心法分离脐血单个核细胞(MNCs),诱导分化为ECFCs,倒置显微镜下观察其原代及传代培养细胞生长状况并通过流式细胞仪、免疫细胞化学、荧光双染色对ECFCs进行鉴定。结果 ECFCs培养过程可出现边界清楚的类圆形单层鹅卵石样细胞集落,细胞增殖能力强,连续传十几代细胞形态稳定,ECFCs高表达内皮系细胞表面抗原,而不表达造血系表面标记;可吞噬Dil-ac-LDL并结合FITC-UEA-1。结论通过简单、可靠的实验方法获得了ECFCs,为进一步探讨其生物学功能及临床应用提供了基础。[ Objective ] To establish a simple method to isolate and culture endothelial colony-forming cells （ECFCs） from human umbilical cord blood in vitro and identify them. [ Methods ] Human umbilical cord blood were collected in sterile conditions and Mononuclear cells were isolated from human umbilical cord blood by density gradient centrifugation , then the cells were differentiated into ECFCs which were identified by flow cytometry, immunocytochemistry and double fluorescent staining. Changes of morphology of primary and passaged cells were observed under inverted microscope . [ Resluts ] ECFCs developed an endothelial shape and appeared clones composed by well-circumscribed monolayers of cobblestone-appearing ceils ; they showed active proliferative capacity and could be passaged for a dozen progenies with stable morphology. ECFCs highly expressed endothelial cell-surface antigens, but not expressed the hematopoietic cell- specific surface antigens ; They could incorporate DiI-acetylated low density lipoprotein （DiI-Ac-LDL）, bind FITC-ulex- lectin （FITC-UEA-I） . [ Conclusion ] ECFCs are acquired by the simple and efficient method which provides basis for further biological function research and clinic application.

关 键 词：内皮克隆形成细胞 诱导培养 细胞鉴定 

分 类 号：R617[医药卫生—外科学]