Abstract Objective：To isolate，culture and identify the bone marrow mesenchymal stem cells-derived neural stem cells（BMSCs-NSCs） in order to observe their growth pattern and differentiation features.Method：Bone marrow mesenchymal stem cells of the Wistar rats were cultured in serum-free medium.Floating neural spheres were formed in serum-free medium，the expression of CD133 and Nestin in the neural spheres was tested by immunohistochemistry.Then floating neural spheres were transferred into serum containing medium for induction differentiation，the expression of GFAP，Map2，β-tubulin Ⅲ and Galc were detected by immunocytochemistry 7 days later. Result：Bone marrow mesenchymal stem cells lived in serum-free medium‘ floating’ and it formed neural sphere.CD133 and Nestin were positive in neural sphere cells according to the results of immunohistochemistry. When these neural sphere cells were transferred into serum-containing medium，they grew adhering to the wall.GFAPMAP-2，β-tubulin Ⅲ and Galc were positive in the cells differentiated from neural spheres. Conclusion：Bone marrow mesenchymal stem cells-derived neural stem cells which growing like a ball and having multipotential differentiation are successfully cultured in serum-free medium with bFGF and EGF.
