Objective To improve the biomechanics properties of completely biological tissue engineered blood vessel (TEBV). Methods Porcine common carotid arteries were used to make the decellularised vascular scaffolds by enzyme digestion, and then the scaffolds seeded with VSMC from canine thoracic aorta were cultured in the vascular culture system, vascular bioreactor with automatic rotation and the effect of mimic pulsatile flow for 3 weeks in vitro, and EC from canine thoracic aorta were seeded subsequently on the scaffolds in the same way and cultured for one more week. After TEBV had been cultured over 4 weeks, the micro-structure of TEBV was examined by histological staining. Meanwhile, the ultrastructure of VSMC and EC in TEBV was also investigated by TEM and SEM. After that, the biomechanics properties of the TEBV were measure by a multi-function mechanical instrument. Results Well-proportioned distribution of VSMC was acquired on the scaffolds in the vascular culture system after 3 weeks. However, there was not such a perfect result even after seeding and culturing for 4 weeks without using the culture system. Endothelization of scaffoldswas achieved in 7 days by using the culture system, but not by using the common pressure perfusion method. The improved TEBV had a histological structure similar to normal physiological one. Normal physiological connections between cells could be found under TEM. Plentiful collagens were also found from seeded VSMCs. The biomechanics properties, including viscoelasticity of TEBV were similar to that of the physiological vessels. Conclusion Both VSMC and EC could adhere or develop into the decellularized scaffolds easier and sooner by using the improved vascular culture system with the effect of automatic rotation and hydromechanics. The improved TEBV presented the better histological structure and biomechanics properties that should be advantageous and significant for TEBV substitute to bear the effect of hydromechanics in vivo.