Influence of basic fibroblast growth factor on differentiation of rabbit bone marrow mesenchymal stem cells into osteoblasts
HU Xiaowei
LI Wei
WEI Dalong
LAO Shan
Abstract：Objective To investigate the effects of basic fibroblast growth factor (bFGF) on differentiation of the rabbit's bone marrow mesenchymal stem cells (BMSCs) into osteoblasts,and to explore the optimal concentration of bFGF.Methods The BMSCs were cultured by the whole blood adherent method and then surface antibody (CD29,CD45 and CD45 by flow cytometry) and osteoblasts (by HE staining and alcian blue staining),and cartilage cells [by alizarin red staining and alkaline phosphatase (ALP) staining] were identified.The 2nd generated BMSCs were cultured with different concentrations (0,1,5,10,50,and 100 ng/mL) of bFGF.detecting alkaline phosphatase (ALP) with PNP colorimetric method and osteocalcin (OC) expression with radioimmunoassay (RIA) method on day 4,7,10 and 14 of culture.Results The positive rate of cell surface antigen CD29 was 92.23％,CD44 was 80.01％,and CD45 was 1.91％.HE staining showed that the cell morphology became polygonal or polygonal and there were multiple protrusions,the black particles could be seen within perinuclear cytoplasm.Alcian blue staining showed thata large number of azure materials diffused.A large number of red calcium nodules were seen by Alizarin red staining.The brown granules and massive precipitation in the cytoplasm were seen by ALP staining,which confirmed the cells were BMSCs with the potential of differentiating into cartilages and osteoblasts.The expression of ALP and OC in the 2nd generation of BMSCs was increased with different concentrations of bFGF as the time passes by.The expression of ALP and OC in BMSCs treated with 50 ng/mg bFGF was higher than that of BMSCs treated with 0,1,5,10 and 100 ng/mL bFGF at the same time (all P ＜0.01).Conclusion The bFGF promotes BMSCs differentiating into osteoblasts which is in a time-dependent manner and the optimal concentration is 50 ng/mg.
Keywords：bone marrow mesenchymal stem cellsbasic fibroblast growth factorosteoblast differentiationrabbit
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 8-11,后插1 )