In this work, bone marrow mesenchymal stem cells and spider silk protein vascular scaffolds were compositely cultured to construct small diameter tissue engineering blood vessel under the dynamic condition, aiming to provide new source of vascular grafts for treatment of cardiovascular disease. Mesenchymal stem cells were cultivated onto the lumen of tubular vascular scaffold under the dynamic culture condition. By means of scanning electronic microscope, HE stain, suture strength test, DNA content detection and hydroxyproline index determination, the formation degree of tissue engineering blood vessel was evaluated. After cultured for 7 days, cells not only fully spread out on the surface of nanofibrous scaffold, but also migrated into fiber interior, showing the vascular scaffold good compatibility. Suture strength of tissue engineering blood vessel was 0.5±0.2 N, which was 29.% of natural blood vessels. The DNA and hydroxyproline content in the tissue engineering blood vessel continually increased over time, and hydroxyproline content on the day 14 and 28 reached 0.6 and 0.2μg/mg respectively, which had significant difference in statistics compared with control group. A kind of small diameter tissue engineering blood vessel was successfully constructed with ideal indexes, which laid foundation for its clinical application.