Objective: To successfully construct the cell-biomaterial scaffold compound with the modified isolated and purified bone marrow mesenchymal stem cells (BMSCs) of SD rats and co-cultured self-made chitosan scaffold in vitro in 3D environment, which laid the foundation for the alternative therapy of transplantation promoting the nerve regeneration.Methods: Three-week old healthy male SD rats were isolated with the bone marrow adherent method.The purification of the cultured third-generation BMSCs were identified with the flow cytometry.The internal structure of the 3D co-cultured BMSCs and the chitosan porous scaffolds prepared with the freeze-drying method in vitro were observed using the scanning electron microscope to measure the diameter of poros, the porosity and the change of growth and morphology of the cells in the scaffold.The proliferation of cells was detected with the MTT method.Results: The third-generation BMSCs were extremely purified.The positive rate of CD29 and CD45 detected with the flow cytometry was 98.49% and only 0.89%, respectively and induced to neuronal differentiation;the chitosan scaffold had an interlinked and uniform porous structure with the porosity of 90%.The BMSCs were adhered closely on the internal wall of the poros and grew well.The chitosan scaffold had no obvious effect on the proliferation of BMSCs.Conclusion: Due to its good structure (diameter and porosity) and biocompatibility, the chitoson scaffold,serving as a carrier, may be realized the promising cell transplantation therapeutic strategy for stroke.