Abstract: Objective To evaluate the effects of genipin cross-linking on biomechanical properties of decellularized trachea treated by detergent enzymatic method (DEM). Methods Thirty tracheal segments from New Zealand white rabbits were divided into 3 groups with 10 in each group. Rabbit tracheas in group A were treated with DEM, group B with genipin cross-linking (1%) after decellularization, and group C were stored in phosphate-buffered saline (PBS) at 4℃ as control group. Biomechanical properties were evaluated by universal tensile testing machine. Results The tensile strength and elastic modulus of group A, B, C were (0.25±0.02) MPa, (0.42±0.09) MPa and (0.28±0.06) MPa and (0.67±0.08) MPa, (1.18±0.31) MPa, (1.19±0.32)MPa, respectively. There was statistically significant difference in elastic modulus between group A and C (P< 0.05), without statistically significant difference in tensile strength (P> 0.05). There were statistically significant differences in elastic modulus and tensile strength between group A and B (P< 0.05), while no statistically significant difference was found in elastic modulus and tensile strength between group B and C (P> 0.05). Conclusion DEM has negative effect on rabbit trachea's mechanical properties, while genipin cross-linking can improve the mechanical properties of decellularized rabbit trachea and the trachea we obtained is mechanically similar to the native trachea.