Using the recently developed surface modification technique, free amino groups have been introduced onto polyester-type polyurethane (PU) scaffolds. The introduction of these free amino groups increases the surface energy and provides a convenient way to further immobilize bioactive species such as gelatin, collagen or chitosan, etc. on the scaffold surface by employing glutaraldehyde as a coupling agent. These modifications are advantageous to enhance cell-material interaction. The culture of human umbilical vein endothelial cells (HUVECs) in vitro proved that the cell proliferation ratio of both the aminolyzed and the biomacromolecules-immobilized PU membranes was improved greatly comparing with the control PU. Scanning electron microscopy and confocal laser scanning microscopy observations displayed that the gelatin-immobilized PU vascular scaffold had formed a monolayer of endothelial intima on its luminal surface after HUVECs were cultured for 6d. Therefore, the aminolysis and the following biomacromolecule immobilization is a promising way to enhance the cell-PU interaction that can accelerate the endothelium regeneration, which is crucial for blood vessel tissue engineering.