Following an in vivo lesion of the rabbit thoracic aorta, endothelial regeneration was studied in silver-stained specimens from en face organ cultures of aortic segments with and without intercostal arteries. The in vivo lesion had been inflicted by an embolectomy catheter. Our results confirmed that endothelium regenerates from preexisting endothelium around the mounths of the intercostal arteries. A conspicuous observation was the orientation pattern of endothelial cells during regeneration. Transmission electron microscopy suggests that the cell cytoskeleton plays a role in the regulation of regeneration. The organ culture technique described, complementary to in vivo and in vitro investigations with cell cultures, seems to be a valuable tool in further investigations on factors involved in the repair of arterial tissue.