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。 Carotid arteries were perfused with 0.1% collagenase digested, a large number of endothelial cells and a small amount of fibroblasts were isolated. After 28 ℃, 1640 + 20% fetal bovine serum with a crude protein content of 350μg / mL In the culture medium, cells grew well, and after 7 days, they were close to monolayer. The original by three times 0. 5% citrate trypsin digestion phase out a small amount of fibroblasts, after 10d, pure endothelial cells, and passed on to the second generation, the second generation of cells by coagulation Ⅷ R factor enzyme assay detected in cultured cells there is coagulation factor Ⅷ R factor The same antigen, with the general characteristics of endothelial cells, was initially identified.