山东医药2016，Vol.56Issue(48)：8-11,后插1,5.DOI:10.3969/j.issn.1002-266X.2016.48.003
碱性成纤维细胞生长因子对兔骨髓间充质干细胞向成骨细胞分化的影响
Influence of basic fibroblast growth factor on differentiation of rabbit bone marrow mesenchymal stem cells into osteoblasts
摘要
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.关键词
骨髓间充质干细胞/碱性成纤维细胞生长因子/成骨分化/兔Key words
bone marrow mesenchymal stem cells/basic fibroblast growth factor/osteoblast differentiation/rabbit分类
医药卫生引用本文复制引用
户小伟,李巍,韦达隆,劳山..碱性成纤维细胞生长因子对兔骨髓间充质干细胞向成骨细胞分化的影响[J].山东医药,2016,56(48):8-11,后插1,5.基金项目
广西中医药民族医药自筹经费科研课题(GZZC15-33). （GZZC15-33）
