Assessment of production of endothelium-derived relaxing factor by cultured porcine cerebral microvessel endothelial cells.

To establish an experimental system to assess the production of endothelium-derived relaxing factor (EDRF) by cultured porcine cerebral microvessel endothelial cells (PCMEC), the effect of PCMEC on thrombin-induced platelet aggregation was studied. For the platelet aggregation cuvettes lined with PCMEC were used. The cuvettes were prepared by seeding PCMEC in gelatin-coated cuvettes at a cell density of 2 x 10(5) cells/ml and culturing for 48 hours. Thrombin-induced platelet aggregation was markedly inhibited in the presence of PCMEC. Indomethacin reduced the PCMEC-dependent anti-platelet aggregatory effect. This PCMEC-dependent anti-platelet aggregatory effect was increased by the addition of bradykinin which stimulates the production of EDRF. These findings suggest that this simple experimental system is useful in assessing the production of EDRF by PCMEC and in examining the effects of various chemicals (or agents) on EDRF production.