鲤血管内皮细胞的分离培养及初步鉴定

SEPARATION CULTURE AND PRELIMINARY IDENTIFICATION OF THE ENDOTHELIAL CELL OF CYPRINUS CARPIO

鲤动脉球腔内灌注０．１％胶原酶消化，分离到大量内皮细胞及少量成纤细胞， ２８℃，在加入蛋白含量为：３５０μｇ／ｍＬ鲤下丘脑粗提液的 １６４０＋ ２０％小牛血清培养液中，细胞生长良好， ７ｄ后，接近单层。原代经三次 ０． ５％柠檬酸胰酶消化逐步淘汰少量成纤维细胞， １０ｄ后，得纯内皮细胞，并顺利传至二代，第二代细胞经血凝ⅧＲ因子酶标检测发现，培养细胞存在血凝ⅧＲ因子相同抗原，具有内皮细胞的一般特征，从而得到初步鉴定。

Many endothelial cells and few fibro-cells were separated fromthe artery ball Cyprinus carpio injected with 0.1% collagense(Ⅰ). At 28℃,in the 1640+10% calf bovine serum medium which was supplemented with 350μg/mL carp hypothulumus-derived growth substance, these cells grew well and the cell monolayers were obtained within 7-days period. By three times digestion of 0.5% trypsin-citric acid solution, the pure endothelial cells were gained after 10 days and they were subcultured to which. The second generation were proved to have ⅧR factor antigen by anti-ⅧR factor assay. These cells were identictied with general biological character of endothelial cells.