During placental development in higher primates trophoblast cells invade maternal blood vessels and migrate along the luminal surface of endothelium. In the present study, the adherence of human cytotrophoblast cells to endothelial cells has been characterized to test the hypothesis that vitronectin receptors (alpha(v) integrins) play a role in intra-luminal trophoblast migration. Adherence was measured using a quantitative fluorescence-based assay and was found to increase in a time-dependent fashion up to about 2 h after which it leveled off. Adhesion was detectable at 4 degrees C but was greatly reduced compared to that seen at 37 degrees C. Adhesion was partially blocked by antibodies against alpha(v)beta3/beta5 integrin, beta1 integrin and by antibodies against P-selectin. Antibodies against beta3 integrin subunits had no effect. Adhesion was reduced by galactose-6-phosphate and fructose-6-phosphate. Flow cytometric analysis revealed alpha(v) integrin on the surface of cytotrophoblast and endothelial cells. Beta1 integrin was detected on the surface of endothelial cells and on cytokine-stimulated cytotrophoblast cells. Beta3 and beta5 integrins were not detected on the surface of either cell type, although beta3 was detected using permeabilized endothelial cells. These results raise the possibility that alpha(v) integrins expressed by both cytotrophoblast cells and endothelial cells, and P-selectin expressed by endothelial cells, may be important in facilitating trophoblast adhesion and migration along the uterine microvasculature.