组织块结合酶消化法培养婴幼儿血管瘤内皮细胞  

Explant combined with trypsin-digestion for culture of endothelial cells from infantile hemangiomas

作　　者：廖洪跃[1] 邢新[1] 欧阳天祥[2] 郭伶俐[1] 李军辉[1] 薛春雨[1] 袁斯明[3] 李鸣[1] 

机构地区：[1]第二军医大学长海医院整形外科,上海200433 [2]上海交通大学医学院新华医院整形外科,上海200092 [3]南京军区总医院烧伤整形科,南京210002

出　　处：《第二军医大学学报》2009年第2期147-150,共4页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(30571928)~~

摘　　要：目的:探讨血管瘤内皮细胞体外培养的方法并观察其生物学特性。方法:收集新鲜手术切除的婴幼儿血管瘤标本,采用组织块结合酶消化法培养内皮细胞,免疫组化EnVision法对培养的细胞进行鉴定;流式细胞仪FITC-CD34检测内皮细胞纯度;相差显微镜下观察血管瘤内皮细胞的生物学特性。结果:共收集血管瘤标本14例,有8例成功培养出内皮细胞,内皮细胞呈现两种形态:多角形和梭形,经免疫组化鉴定为内皮细胞;培养的细胞中CD34+细胞数为76.28%;血管瘤内皮细胞有明显的"管腔形成"。结论:组织块结合酶消化法能成功培养较纯的婴幼儿血管瘤内皮细胞,培养的内皮细胞具有某些生物学特性。Objective： To explore a novel method for cultivation of the endothelial cells（EC） from infantile hemangiomas （IH） in vitro and observe the biological character of cultured endothelial cells. Methods： Fresh operative specimens were obtained from infantile hemangiomas. The endothelial cells of IH were cultured by explant combined with trypsin-digestion technique. Immunohistochemical staining of EnVision method was carried out to identify the cultured cells. The purity of endothelial cell was examined by flow cytometry analysis of FITC-CD34. The biologic characters of endothelial cells were observed under inverted phase contrast microscope. Results： Endothelial cells were successfully cultured from 8 IH specimens of all the 14 explants. The morphology of cultured endothelial cells included two types：polygonal cells and fusifourm cells. The cultured cells were homogenously positive for EC markers-vWF or CD34,indicating their EC origin. Tube formation was found in endothelial cells of IFI. The proportion of CD34^- cells was 76.28% in cultured endothelial cells as detected by flow cytometry analysis. Conelusion：ECs can be successfully isolated and cultured from infantile hemangiomas by explant combined with trypsin-digestion technique; the cultured cells have some characters of endothelial cells.

关 键 词：血管瘤 婴幼儿 内皮细胞 细胞培养 组织块结合酶消化法 

分 类 号：R732.2[医药卫生—肿瘤]