Osteogenic characteristics of rabbit bone marrow mesenchymal stem cells under three kinds of culture conditions

Objective To investigate osteoblast phenotype differentiation of rabbit bone marrow mesenchymal stem cells(BMSc) under three different culture conditions.Methods BMSCs were cultured in DMEM/F12 medium,induction medium and co-culture medium with fresh bone particles to compare the their osteoblast phenotype differentiation and investigate alkaline phosphatase(ALP) activities,and expressions of osteocalcin and type Ⅰ collagen.Results ALP activities and osteocalcin in BMSCs in induction culture and co-culture media were significantly higher than those in DMEM/F12 medium at the same period(P0.01).The ALP activities and osteocalcin in BMSCs in induction culture medium were higher than those in BMSCs in co-culture medium.In BMSCs in induction culture and co-culture media,immunohistochemical staining of typeⅠcollagen and osteocalcin was positive.Immunohistochemical staining of typeⅠcollagen and osteocalcin in control group was feebly positive and negative respectively.Expressions of typeⅠcollagen of BMSCs that were passed for one and two weeks were significantly lower than those in induction cultured and co-cultured MSCs. Conclusion BMSCs cultured in DMEM/F12 medium and co-cultured with fresh bone particles both have significant osteogenic capacity.But induction culture condition can result in high level of ALP activities and osteocalcin.