Construction of the stable monolayer of endothelial cells onto physicochemically modified polymeric surFace is one of the appropriate method to develop the small caliber vascular graft with the long-term patency. In this study, we constructed the monolayer of endothelial cells on the fibronectin rind the extracellular matrix-coated polyurethane surface derived from human fibroblast cells. To elucidate the adhesion strength of endothelial cells on the extracellular matrix-coated polyurethane, a laminar flow chamber apparatus was developed to exposure the shear stress on the apical membrane of ondothelial cells. Endothelial cells show the strongest adhesion after two days of seeding onto the fibronectin-coated polyurethane surface, whereas endothelial cells on the extracellular matrix derived from the human flbroblast cells show the minimal doubling time of cellular growth.