Objective The paper discussed the preparation of human acellular amniotic membrane (HAAM), morphology and ability of releasing nitrous oxide (NO) and the adhesion after endothelial cells seeding on HAAM so as to offer an improved approach for preventing in-stent restenosis by making HAAM cover to modify intravascular stent and seed endothelial cells on it. Methods HAAM was prepared by removing the original epithelium by pancreatic enzymeas the substrate for human umbilical vein endotholical cells (HUVECs) seeded on it and its morphology, ability of releasing NO and the adhesion were examined. Results HUVECs were successfully seeded and could proliferate on HAAM, the monolayer was formed after cultivating 67 days. Their morphology, ability of releasing NO and the adhesion showed better than that of the endothelial cells of control. Conclusion HAAM can be used as a substrate for cultivating HUVECs and improving its function.