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Normal and von Willebrand(vWd) porcine aortic endothelial cells(EC) have been maintained and subcultured for10-12 months(14-18 passages) without any apparent change in characteristic growth morphology or ultrastructure. Immunofluorescence staining ofthese EC for ristocetin-Willebrand factor(RWF), using monospecific rabbit antiporcine RWF(aRWF), showed differences in the extent and nature of intercellular staining and the apparent absence of extracellular EC associated RWF-containing material(filaments)in vWd cells. Platelet-endothelial(damaged) interaction was decreased in normal EC cultures by pretreatment of the cultures with aRWF, no platelet interaction was seen with untreated vWd EC. These cultures were used as an in vitro model system(optimal pH, divalent cations, protein, exposure time and rotation speed)for studying the nature, extent and differencesin the platelet-microfi1ament interaction in normal and vWd EC. Preliminary data on the microfilament site of attachment of washed platelets or antibody-coupled beads, the interaction of platelets with matrix-bound purified RWF and the presence of immunologically identifiable RWF-containing filaments in normal and vWd EC suggest a possible role for endothelial-bound RWF in platelet-vessel wall interaction.

thrombosis and haemostasis

Cultured Normal and von Willebrand Porcine Aortic Endothelial Cells-An in Vitro Model for Studying the Nature and Site of Platelet-Endothelial Component Interaction