[骨形态发生蛋白-2和碱性成纤维细胞生长因子-2对小鼠间充质干细胞体外成骨细胞分化的影响]

[Effects of BMP-2 and FGF-2 on osteoblast differentiation of murine MSCs in vitro].

作者信息

Zhang Wei-xi, Chen Song-lin, Yao Xiao-li, Lu Xi-lin, Zhou Chen, Leng Yan, Shi Xingming

机构信息

Department of Neurology, First Hospital of Sun Yat-sen University, Guangzhou 510080, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Nov;24(11):1062-5.

PMID:18992192

Abstract

AIM

To study the effects of BMP-2 and FGF-2 on osteoblast differentiation of murine MSCs in vitro.

METHODS

The bone marrow cells were collected from 3-18 month old C57BL/6J mice (50 mice), and they were isolated, enriched and expanded using bone marrow adherent culture, and then purified by immunomagnetic microbeads. At last they were identified as mesenchymal stem cells (MSCs). After the MSCs were cultured adherently for 24 hours, 100 microg/L BMP-2 and 0.5 nmol/L FGF-2 were added into osteogenic media separately for 7, 14 and 21 days, alkaline phosphatase (ALP) staining, alkaline phosphatase activities, Vonkossa staining, and Alizarin red S staining were performed step by step. Osteoblast differentiation marker genes including Runx2/cbfa1, ALP, collagen-1, and osteocalcin were investigated by RT real time PCR.

RESULTS

ALP activities, calcium deposition and the osteoblast differentiation marker genes of BMP-2 group were markedly higher than those of osteogenic group. The mRNA of Runx2/cbfa1, ALP, collagen-1, and osteocalcin of BMP-2 group was highly expressed than that of osteogenic group. It was the same with FGF-2 group, but less evident than BMP-2 group.

CONCLUSION

BMP-2 and FGF-2 can induce murine MSCs to differentiate into osteoblast at different degrees.

摘要

目的

研究骨形态发生蛋白-2（BMP-2）和碱性成纤维细胞生长因子-2（FGF-2）对小鼠间充质干细胞（MSCs）体外成骨细胞分化的影响。

方法

从3至18月龄的C57BL/6J小鼠（50只）中采集骨髓细胞，采用骨髓贴壁培养法进行分离、富集和扩增，然后通过免疫磁珠进行纯化，最终鉴定为间充质干细胞。将MSCs贴壁培养24小时后，分别向成骨培养基中加入100μg/L BMP-2和0.5nmol/L FGF-2，分别培养7、14和21天，依次进行碱性磷酸酶（ALP）染色、碱性磷酸酶活性检测、Vonkossa染色和茜素红S染色。通过RT实时PCR检测成骨细胞分化标志物基因，包括Runx2/cbfa1、ALP、胶原蛋白-1和骨钙素。