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

发布时间：2018-11-21 11:37

【摘要】：目的观察碱性成纤维细胞生长因子(b FGF)对兔骨髓间充质干细胞(BMSCs)向成骨细胞分化的影响,并探讨其最佳作用浓度。方法采用体外全血贴壁法培养兔BMSCs,进行表面抗原(CD29、CD44、CD45,采用流式细胞术)及成软骨(HE染色和阿尔新蓝染色)、成骨[茜素红染色及碱性磷酸酶(ALP)染色]分化潜能鉴定。取第2代BMSCs,加入含0、1、5、10、50、100 ng/m L b FGF的成骨诱导液,分别于培养第4、7、10、14天采用PNP比色法检测ALP表达,采用放射免疫分析法检测骨钙素(OC)表达。结果细胞表面抗原CD29阳性表达率为92.23%,CD44阳性表达率为80.01%,CD45阳性表达率为1.91%;HE染色见细胞形态为多角形或多边形,有多个凸起,核周细胞质内可见黑色颗粒,阿尔新蓝染色可见细胞间大量天蓝色物质弥漫分布;茜素红染色可见大量红色钙结节,ALP染色可见细胞质中出现棕褐色的颗粒和块状沉淀;证实分离培养的细胞为BMSCs,并具有成软骨和成骨分化潜能。随着作用时间的延长,BMSCs经不同浓度b FGF作用后ALP及OC表达均呈升高趋势。相同作用时间下,BMSCs经50 ng/m L b FGF作用后ALP及OC表达均高于0、1、5、10、100 ng/m L b FGF作用后(P均0.01)。结论 b FGF能促进BMSCs向成骨细胞分化,并呈时间依赖性;其最佳作用浓度为50 ng/m L。
[Abstract]:Objective to observe the effect of basic fibroblast growth factor (b FGF) on the differentiation of rabbit bone marrow mesenchymal stem cells (BMSCs) into osteoblasts by (BMSCs), and to explore the optimal concentration of BFGF. Methods Rabbit BMSCs, was cultured with whole blood adherent method in vitro for surface antigen (CD29,CD44,CD45,) and cartilage (HE staining and new blue staining). The differentiation potential of osteogenesis [alizarin red staining and alkaline phosphatase (ALP) staining] was identified. The second generation of BMSCs, was added into the osteogenic inducer containing 0 1, 1, 5, 10, 10, 50 ng/m L b FGF. The expression of ALP was detected by PNP colorimetry on the 4th day of culture, and the expression of osteocalcin (OC) was detected by radioimmunoassay (RIA). The expression of osteocalcin (OC) was detected by radioimmunoassay (RIA). Results the positive expression rate of cell surface antigen CD29 was 92.23 and the positive rate of CD44 was 80.01 and the positive rate of CD45 was 1.91. HE staining showed that the cells were polygonal or polygonal, with multiple bulges, black particles could be seen in the cytoplasm around the nucleus, and a large number of sky blue substances were diffuse in the cells. Alizarin red staining showed a large number of red calcium nodules, and ALP staining showed brown granules and massive precipitates in the cytoplasm, which confirmed that the isolated and cultured cells were BMSCs, and had the potential of cartilage formation and osteogenic differentiation. The expression of ALP and OC in BMSCs treated with different concentrations of b FGF increased with the prolongation of time. At the same time, the expression of ALP and OC in BMSCs treated with 50 ng/m L b FGF was significantly higher than that after treatment with 10 100 ng/m L b FGF (P 0.01). Conclusion b FGF can promote the differentiation of BMSCs into osteoblasts in a time dependent manner, and its optimal concentration is 50 ng/m / L.
【作者单位】：广西医科大学第一附属医院;
【基金】：广西中医药民族医药自筹经费科研课题(GZZC15-33)
【分类号】：R329.2

【相似文献】