Objective To compare the biological and biomechanical characteristics of decellularized bovine jugular venous tissue-engineered valved conduit scaffolds with that of fresh bovine jugular veins. Methods Fortyeight fresh bovine jugular veins were divided into control group and experimental group with random number table method, 24 veins in each group. There were fresh bovine jugular veins in control group, decellularized bovine jugular veins in experimental group. The veins of experimental group were treated with sodium deoxyeholate plus Triton-X-100 to decellularize the cells in valves and vessel walls. The thickness, water absorption rate, water maintenance rate, destroying strength, stretch rate of valves and vessel walls in two groups were detected. Results The endothelial cell and fibroblast of valves and vessel walls in experimental group were completely decellularized, no cell fragments were retained within the matrix scaffold; collagen fiber and elastin fiber had been preserved with intact structure and wavily arrayed; deoxyribonucleic acid content of valves and vessel walls in experimental group were decreased by 97.58%, 97.25% compared with that of control group. The thickness, water absorption rate and water maintenance rate of valves and vessel walls in experimental group were lightly increased than those of control group, but there were no significant differences between them (P 〉 0. 05). There were no significant differences in destroying strength and stretch rate of valves and vessel walls between two groups (P〉0. 05). Conclusion Decellularized bovine jugular vein scaffold has stable biological and biomechanical characteristics and it may be ideal natural fibrous matrix for developing the tissue-engineered valved conduit by host recellularization.