Bone tissue engineering is considered as one of the most promising way to treat large segmental bone defect. When constructing bone tissue engineering graft in vitro, suitable bioreactor is usually used to incubate cell-scaffold complex under perfusion to obtain bone tissue engineering graft with good repair efficiency. However, the theoretical model for growth rate of single cell (especially for stem cell) during this process still has many defects. The difference between stem cells and terminally differentiated cells is always ignored. Based on our previous studies, this study used self-made perfusion apparatus to apply different modes and strengths of fluid shear stress (FSS) to the cells seeded on scaffolds. The effects of FSS on the proliferation and osteogenic differentiation of mesenchymal stem cells (MSCs) were investigated. The regression analysis model of the effect of FSS on the single-cell growth rate of MSCs was further established. The results showed that 0.022 5 Pa oscillatory shear stress had stronger ability to promote proliferation and osteogenic differentiation of MSCs, and the growth rate of a single MSC cell under FSS was modified. This study is expected to provide theoretical guidance for optimizing the perfusion culture condition of bone tissue engineering grafts in vitro.