Objective To explore the effects of Sertoli cells on the induced adipogenic, osteogenic and neuroblastic differentiation of bone marrow mesenchymal stem cells (BMSCs). Methods BMSCs were isolated and purified with the density gradient centrifugation method, Sertoli cells were isolated with two-step enzyme digestion;The adipogenic, osteogenic and neuroblastic differentiation of P3/P4 BMSCs were induced. Sertoli cells were co-cultured with BMSCs and then the morphological change of BMSCs were observed. Results After adipogenic induction, the lipid droplets that have strong refraction in the cytoplasm were observed by oil red O staining. The intracellular lipid droplets shrunk and the orange stained lipid droplets were hard to observe after adding Sertoli cells; After osteogenic induction, calcium nodules were shown by alizarin red S staining. More clearer calcium nodules were observed and the red-staining was stronger after adding Sertoli cells. No morphological change was observed on BMSCs before and after adding Sertoli cells. Conclusion Sertoli cells could promote BMSCs’ osteogenic differentiation, inhibit their adipogenic differentiation and have no effect on their neuroblastic differentiation.