Endothelial cell seeding has been successful in reducing the thrombogenecity of prosthetic vascular grafts in animal models, but results from clinical trials have been largely disappointing. These poor results have been associated with poor graft coverage in immediate seeding trials, and failure of cell culture in staged procedures. These problems could be largely overcome by utilising a bank of allogeneic endothelial cells, providing an ever ready supply. However, one potential pitfall with this technique would be the possibility of a rejection response following transplantation. OBJECTIVE To study the effects of prolonged tissue culture, on the ability of endothelial cells to generate an immune response. RESULTS The immunogenecity of human umbilical vein endothelial cells was measured using the mixed lymphocyte endothelial reaction. It was demonstrated that prolonged tissue culture significantly reduced the immunogenecity of the cells, from a mean of 7261 cpm (S.E. +/- 243, n = 3) for cells of subculture 3, to 5478 cpm (+/- 156, p = 0.04) for cells of subculture 7 (p = 0.04, Wilcoxon paired rank test), but did not significantly impair morphology or antithrombotic function. CONCLUSIONS This study provides evidence that prolonged tissue culture provides morphologically and functionally intact, immunomodified endothelial cells which may potentially be used in seeding prosthetic vascular grafts.