Primary study of cell phenotype of isolated and cultured canine bone marrow mesenchymal stem cells in vitro

    OUYANG Hui, LIU Weiª²Yong, WEI Xiangª²Yang, GU Chunª²Hu, SU Xueª²Jia, ZHENG Qiª²

    Jun, LIU Xingª²Guang

    Department of Cardiovascular Surgery, Xijing Hospital, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To observe the phenotype changes and biological characteristics of naturally differentiated canine bone marrow mesenchymal stem cells (BMSCs) and differentiated after induced with bFGF in vitro. METHODS: BMSCs were isolated and purified from the canines bone marrow by Percoll density gradient centrifugation and by adhering to the culture plastic. After successive culture and amplification, the growth curve was drawn. The morphology was observed under phase contrast microscope and some antigens were examined by immunohistochemistry stain. RESULTS: SH2, Vimentin, ¦Áª²SMA, collagen ¢ñ and ¢ó were all positively expressed in naturally differentiated BMSCs, with the positive percentages (95.1¡À1.7)%, (64.4¡À2.2)%, (78.7¡À0.9)%, (78.3¡À1.4)% and (66.8¡À0.8)%, respectively. CD34, ¢ø antigen and laminin displayed negative reaction. The induced and differentiated cells showed positive staining of lamininn [(50.2¡À2.5)%]. The positive expression of ¦Áª²SMA decreased to (42.3¡À1.6)% and that of vimentin increased to (86.2¡À2.4)%. CONCLUSION: The phenotypes and biological characteristics of naturally differentiated myofibroblast with canine BMSCs show that they are more active than those of fibroblasts and more suitable to be used as seedª²cells in constructing tissue engineering heart valve (TEHV). ......