论文部分内容阅读
目的 建立一个稳定的人血管畸形内皮细胞体外培养体系。方法 取人静脉畸形组织块接种于铺有明胶底层的培养瓶中进行原代及传代培养。通过早期去除组织块、机械刮除及酶消化法纯化内皮细胞。对培养的细胞进行形态学观察及免疫组化染色等一系列细胞定性研究。结果 获取的内皮细胞可连续传代 4～ 5代 ,成活 4 0～ 5 0d ;生长于玻璃、塑料上的细胞呈铺路石状 ;生长于明胶底层上的细胞可形成毛细血管样结构 ;电镜下可见细胞具有Weibel Palade小体特征性超微结构 ;CD3 4 及vWF免疫组化染色阳性 ,α SMA染色阴性。结论 应用此培养方法可获得纯化的血管畸形内皮细胞 ,且细胞可连续传代培养。培养的血管畸形内皮细胞可用于体外研究血管畸形的生物学行为。
Objective To establish a stable human vascular malformation endothelial cell culture system in vitro. Methods Human venous malformations were seeded into gelatin-coated flasks for primary culture and subculture. Endothelial cells were purified by early removal of tissue pieces, mechanical scraping and enzyme digestion. A series of cell qualitative studies were carried out on the cultured cells for morphological observation and immunohistochemical staining. Results The endothelial cells were passaged for 4 to 5 generations continuously and survived for 40 days to 50 days. Cells growing on glass and plastic were paved stone. Cells grown on the bottom of gelatin could form capillary-like structures. The cells had the characteristic ultrastructure of Weibel Palade bodies; the immunohistochemical staining of CD34 and vWF was positive and the negative staining of α SMA. Conclusion Purified vascular malformation endothelial cells can be obtained by this culture method, and the cells can be subcultured continuously. Cultured vascular malformations endothelial cells can be used to study the biological behavior of vascular malformations in vitro.