A method for analysis of adhesion of platelet to cultured HUVECs under conditions of flow

To study the interaction of platelets to cultured human umbilical vein endothelial cells (HUVECs) under shear flow conditions in vitro, a dynamic platelet thrombus model based on the endothelial. cell culture and microfluorescence technique was developed. A parallel plate flow chamber in which the flow is two-dimensional and fully developed laminar was adopted in this study. After injured by H2O2 and co-incubated with platelet rich plasma (PRP), HUVECs monolayer was exposed to shear stress rate ranging from 50 to 800 sec(-1) respectively for 10 minutes. By a series of optical section images sampled by Laser Scan Confocal Microscope (LSCM), adhesion of platelets with HUVECs in different conditions was quantitated. In pharmacological experiments, Aspirin were added in PRP to testify its antithrombus effect in now conditions. The results show the endothelial cell-platelet thrombus was successfully formed in vitro. After the action of Aspirin, adhesion area of thrombus decreased. The study indicates this thrombus model in vitro and the corresponding quantitative analysis system is convenient and useful in the studies of endothelial cells-platelets interaction and mechanism of thrombosis and atherogeniesis. It can also be used to evaluate the effects of antithrombus drugs.