重组Osf2定向调控骨髓间充质干细胞向成骨细胞分化

Study on the definitive differentiation from bone marrow mesenchymal stem cells to osteoblast regulated by recombined Osf2

目的研究成骨细胞特异性因子(Osf2)调控骨髓间充质干细胞(MSCs)定向分化为成骨细胞(OB)的分子机制。方法用Percoll密度梯度离心法分离获得大鼠的MSCs,通过细胞免疫化学、免疫荧光、透射电镜及流式细胞仪等方法进行鉴定。构建真核表达载体pcDNA3.1(+)/Osf2,通过脂质体系统转染传代培养后的MSCs,用Northern Blot检测三种OB细胞外基质蛋白mRNA的表达情况。结果转染后的MSCs都表达OB特异性基质蛋白骨钙素、骨桥接蛋白和I型胶原。结论Osf2在转录水平定向调控MSCs分化为成骨细胞,可应用于骨质疏松、骨缺损等疾患的基因治疗。

Objective To investigate the molecular regulation mechanisms of Osf2 to differentiation of mesenchymal stem cells（MSCs） to osteoblast. Methods MSCs come from bone marrow of rats with the method of percoll density gradient centring, and were evaluated with combination methods, including immunochemistry, immunofluorescence, transmission electron microscope and flow eytometry analysis. Subcultured MSCs were tranafected by recombinant eukaryon expression vector, pcDNA3.1 （ ＋ ）/Osf2, under the transfection system of liposomes. Northem Blot was used to detect the mRNAs of extra cellular matrix protein（ECMP） of osteoblast. Results Expression of osteocalcin, osteopontin, type I collagen mRNAs were detected in transfected MSCs. Conclusion Osf2 can regulate differentiation from MSCs to osteoblast on transcription level, and can be applied to treat orthopedic diseases, shueh as osteoporosis, defect of bone, etc.