Aim To study the adhesion and retention of endothelial cells in different conditions, such as different shear stress and substrates, and gene trasfected or not, with the parallel plate flow chamber. Methods After treated with different substrates, endothelial cells were cultured on the substrates, and were processed by the same size shear stress. Or endothelial cells were processed by different size shear stress which cultured on the same substrate. Results Cells retention ratio was higher in low shear stress than high shear stress although both of them were in normal shear stress. Cells retention ratio was high in poly-l-lysine and fibronectin than in collagen Ⅳ. There was no effect on retention of endothelial cells in poly-l-lysine that gene transfected into endothelial cells. Conclusion Different size shear stress and substrates effect the retention of endothelial cells.