Effects Of Basic Fibroblast Growth Factor On Osteoblast Differentiation Of Rabbit Bone Marrow Mesenchymal Stem Cells

Objective:To observe the biological characteristics and the differentiation potential of the rabbit bone marrow mesenchymal stem cells, and then investigated the effects of basic fibroblast growth factor(bFGF) on osteoblast differentiation of the rabbit's bone marrow mesenchymal stem cells (BMSCs) in vitro.Methods:The rabbit bone marrow-derived MSCs were isolated and cultured from rabbit bone marrow by the bone marrow different adherent method in vitro,then observed under the inverted microscope morphology and growth of BMSCs. The 2nd generated BMSCs which have detected by flow cytometry and induced to osteoblast and cartilage cells, was identified by HE staining and Alcian blue staining for cartilage cells, and Alizarin red staining and ALP staining for osteoblast cells. The 2nd generated BMSCs was induced to osteoblast cells with different concentrations(1,5,10,50,100ng/mL)of bFGF and blank group for 14 days, then analyse the osteoblasts performance differences in 6 groups by detecting the expression of Alkaline phosphatase (ALP) and osteocalcin (OC).Results:The 3th generated BMSCs which cultured in the different adherent method in vitro was high expression of CD29(92.23%) and CD44(80.01%),low expression of CD45(1.91%) by flow cytometry, and also can be induced by specific way to cartilage cells and osteoblast. After cartilage induction, HE staining showed that cytoplasm existed black particles, Alcian blue staining showed the existence of blue acid mucopolysaccharide. After osteogenetic induction, mineralized nodules were detected by alizarin Bordeaux staining, and the induced-cells were strong positive to alkaline phosphates staining. The bFGF is effective in promoting osteoblastic differentiation in inducing osteogenesis of BMSCs(P<0.01), and the expression of ALP and OC in experimental group were significantly higher than the blank group. The concentration of 50ng/mL of bFGF could significantly promote BMSCs'osteoblast differentiation (P<0.01).Conclusion:The bone marrow different adherent method can be effectively purified BMSCs in vitro. The bFGF is an effective induction factor to promote osteoblast differentiation of BMSCs and depending on its proper concentration.