Abstract: Many endothelial cells and few fibro-cells were separated from the artery ball Cyprinus carpio injected with 0.1% collagense(I). At 28, in the 1640+10% calf bovine serum medium which was supplemented with 350μg/mL carp hypothulumus-derived growth substance, these cells grew well and the cell monolayers were obtained within 7-days period. By three times digestion of 0.5% trypsin-citric acid solution, the pure endothelial cells were gained after 10 days and they were subcultured to which. The second generation were proved to have VIIIR factor antigen by anti-VIIIR factor assay. These cells were identictied with general biological character of endothelial cells.