血管内皮细胞体外培养及生物学特性的探讨

目的　分离培养脐静脉内皮细胞 ,鉴定其生物学特性。方法　采用 0 2 %胰蛋白酶行脐静脉两次酶灌注消化法获取内皮细胞。分别在倒置相差显微镜、透射电镜下观察 ,SP免疫组化法行Ⅷ因子抗原鉴定。观察血管内皮细胞体外培养的生长情况 ,并绘制细胞生长曲线。结果　 (1)经改良的胰蛋白酶两次消化法获取较高的活细胞百分数 ,且细胞纯度较高。 (2 )透射电镜下发现多数细胞间存在紧密连接结构 ,胞膜的一侧贴有基膜样成分。 (3)血管内皮细胞体外培养融合成单层后 ,形成类似体外血管内皮的内衬结构。结论　透射电镜下细胞特征可作为鉴定血管内皮细胞的辅助手段。

The Biologic Characteristics of Human Vein Endothelial Cells Cultured in Vitro

Objective In view of the similarity between endothelial cell of umbilical vein and endothelial cell of cavernous hemangioma in biological characteristics, we have separated the endothelial cells from umbilical vein and cultured in intro to distinguish their biological characteristics, so that we could establish the base of drug intervention experiment of endothelial cells in vitro. Methods We obtained the endothelial cells by enlymatic perfusion digestion of umbilical vein twice using trypsinase (0 2%). We observed the morphologic characteristics of endothelial cells under the inverted microscope and fuloroscopy, and indentified the factor Ⅷ antigen by SP immunohistochemical method. We draw cell growth curve by observing the growth of endothelial cells cultured in vitro. Results (1)Well purified HVEC with few toxic effects were obtained by two staged digestion of trypsinase. (2)HVEC in confluent primary culture reproduce the epithelioid organization of the vascular lining in situ. (3) The marked structure of junctional integrity and substratum attached to one side of cells may be a supplement to identify HVEC. Conclusion The characteristics of the cells under fuloroscopy can be used to identify HVEC.