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Characterization and properties of cultured human von Willebrand umbilical vein endothelial cells.

Endothelial cells (ECs), designated here as vWd ECs, were obtained from the umbilical vein of a female child, subsequently diagnosed as having von Willebrand’s disease (vWd). ECs were obtained from human umbilical veins (3 normals, 1 vWd) by mild collagenase treatment and cultured on human fibronectin (FN) coated culture surfaces in Medium 1 99 containing 20% heat-deactivated fetal calf serum and bovine EC growth factor (ECGF). These cells were characterized as ECs by their typical growth characteristics, the presence of immunologically identifiable factor-VIll-related antigen (F.VIIIR:AG) and the production of ristocetin-Willebrand factor (F.VlllR:WF) activity (ristocetin-induced platelet aggregation). The ECs in postconfluent vWd cultures appeared somewhat larger and showed an increased tendency to become detached from the culture surface, as compared to postconfluent normal cultures. In comparison with normal ECs. vWd ECs showed a different pattern of immunofluorescencestaining for F.VlllR:AG and lower levels of this factor in the culture medium. In 72-hr. normal ECs also produced 3-4-fold more F.VIIIR:WF activity in their media than vWd ECs. Finally, normal culture-produced extracellular matrix material,