We describe the preparation of segmented polyurethane (SPU) films with round micropores and present a quantitative assay method of endothelial cell (EC) migration through micropores of and growth on microprocessed SPU films as an in vitro model of transmural endothelialization in open-cell-structured small-diameter vascular grafts. The micropored films, pores of which ranged from 9 to 100 microns in diameter, were microfabricated using an excimer laser. Time-dependent processes of EC ingrowth through micropores of SPU films with different pore sizes, which have a confluent monolayer sheet on one face and are cell free on the other, and subsequent endothelialization were quantitatively studied. The circular cellular sheet centered at the micropores expanded as incubation proceeded. Markedly retarded migration was found for the smallest pore size (9 microns in diameter). The larger the pore, the higher was the endothelialization rate. The endothelialization characteristics were studied on multiply microspored films of different pore sizes and densities, each of which was prepared so as to provide a fixed total pore area per unit area (0.01 mm2 per mm2). The highest endothelialization rates in an early incubation period were found on films with microspores between 18 and 50 microns in diameter.