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A novel cell line named B-1085 with morphological and physiological stability was established from the tissue cultured cells of the human ovary, which produced high level of plasminogen activator (PA) into the cultured medium.B-1085 was subcultured under the following conditions: MEM medium containing 10% FCS, 2mM glutamine and 300ppm penicillin with 5×105 cells/10ml of medium/25cm2-flask, incubated at 37°C, 5% CO2 for 7-10 days. This cell line could be cultured on microbeads with a good adhesion and growth secreting high concentration of PA into the medium.PA produced by B-1085 was investigated on its enzymatic identification. This PA was successfully purified using high performance liquid chromatography with zinc chelate-5PW column resulting in a single band (55K) on SDS-polyacrylamide gel electrophoresis. Enzymography revealed that 55K protein band had a fibrinolytic activity and no other active band was detected. Immunological study showed that the PA cross-reacted with anti-UK antibody but not with anti-t-PA one. These results suggest that the PA was similar to UK in the the point of molecular weight and immunological property. On the other hand, this PA demonstrated high affinity to fibrin, indicating t-PA type characteristic.

Studies on plasminogen activator produced by the cultured cell line from the tissue of human ovary.