Emeis J J, Edgell C J
Gaubius Institute TNO, Leiden, The Netherlands.
Blood. 1988 Jun;71(6):1669-75.
The fibrinolytic characteristics of the endothelial hybrid cell line EA.hy 926, established by fusing a human umbilical vein endothelial cell with a human carcinoma cell line, were studied. The hybrid cell line produced large amounts of tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor type 1, and a small amount of urokinase. All plasminogen activator present in conditioned medium was complexed with inhibitor because the cells secreted plasminogen activator inhibitor in excess over plasminogen activator and no activator activity was detectable in conditioned media by direct activity assays. t-PA activator activity was, however, demonstrable in conditioned media after treatment with sodium dodecyl sulfate, in agreement with t-PA antigen determinations. Increased plasminogen activator inhibitor activity could be induced by incubating the cells in the presence of endotoxin or microtubule inhibitors, whereas increased t-PA activity could be induced by microtubule inhibitors. Interleukin-1 had no effect. The fibrinolytic characteristics of the hybrid cell line were stable for at least 30 passages. The perpetual human hybrid cell line EA.hy 926 therefore may be a useful tool for the study of fibrinolysis in cultured endothelial cells.
通过将人脐静脉内皮细胞与人癌细胞系融合建立的内皮杂交细胞系EA.hy 926的纤溶特性进行了研究。该杂交细胞系产生大量组织型纤溶酶原激活物(t-PA)、1型纤溶酶原激活物抑制剂,并产生少量尿激酶。条件培养基中存在的所有纤溶酶原激活物都与抑制剂结合,因为细胞分泌的纤溶酶原激活物抑制剂超过了纤溶酶原激活物,通过直接活性测定在条件培养基中未检测到激活物活性。然而,在用十二烷基硫酸钠处理后的条件培养基中可证明t-PA激活物活性,这与t-PA抗原测定结果一致。在内毒素或微管抑制剂存在下培养细胞可诱导纤溶酶原激活物抑制剂活性增加,而微管抑制剂可诱导t-PA活性增加。白细胞介素-1无作用。杂交细胞系的纤溶特性在至少30代内保持稳定。因此,永久性人杂交细胞系EA.hy 926可能是研究培养内皮细胞纤溶作用的有用工具。