The osteoblasts were expanded in large-scale by microcarrier suspension culture in rotating wall vessel bioreactor(RWV), then the biological functions of the cells were detected by histomorphometry and the cells were seeded at 2×10⁶ cells/ml and 1×10⁶ cells/ml onto scaffolds respectively to culture for 2 weeks. The biological properties of the fabricated bone tissue were detected by inverted microscope, scanning electron microscope (SEM), alkaline phosphatase (ALP), von-Kossa staining on mineralized nodules and AO/EB fluorescence staining. Meanwhile the cells' metabolism of nutrients was monitored and analyzed during the whole culture process. The bone tissue fabricated in RWV with two different seeded cell densities grew well. Osteoblasts in the scaffolds secreted much collagen fibers and formed mineralized nodules and new osteoid tissue. It can be conclueled that: With the stress stimulation inside the fluid in the RWV, the active expression of ALP can be increased, the formation of mineralized nodules can be accelerated. The rapid proliferation and differentiation of osteoblasts are possible and the three-dimensional fabrication of engineered bone could be realized.