Abstract: In order to explore which way can be more efficient and faster to isolate, culture and identifiy the canine bone marrow mesenchymal stem cells (BMSCs) in vitro, the BMSCs were isolated in vitro by whole bone marrow differential velocity adherent and density gradient centrifugation. Immunohistochemistry and flow cytometry were used to examine the surface markers of the cells, and induced their differentiation into osteoblasts and adipocytes. The results showed that canine BMSCs were successfully cultivated by whole bone marrow differential velocity adherent and density gradient centrifugation. Compared with the former, primary cells from the latter method was more uniform after cultivation, and cost shorter time for primary cells proliferation into full confluency along with higher survival rate. The growth curves of undifferentiated cells in P3 and P8 were similar from the both methods. Immunohistochemistry and flow cytometry showed that CD105, CD90 and CD29 were expressed in the cells and could be induced into osteoblasts and adipocytes, respectively, while CD34 and CD31 were not expressed. These results indicated that canine BMSCs could be isolated and cultivated by the both methods, and the density gradient centrifugation method was better compared to the whole bone marrow differential velocity adherent.

Key words: canine, bone marrow mesenchymal stem cell, isolation and cultivation