摘要
目的:携带骨形态发生蛋白2(bone morphogenetic protein-2,BMP-2)基因的慢病毒载体感染大鼠骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)后,观察BMP-2对BMSCs成骨作用的影响。方法:原代培养大鼠BMSCs并进行传代及鉴定,用携带BMP-2的慢病毒感染第3代BMSCs后,进行骨向诱导,通过茜素红染色观察钙沉积。观察细胞黏附能力及对成骨标志分子骨桥蛋白(OPN)、骨钙素(OCN)、胶原蛋白Ⅰ(Col-Ⅰ)及smad(drosophila mothersagainst decapentaplegic protein)表达的影响。采用SPSS11.0软件包对数据进行统计学分析。结果:①原代培养出大鼠骨髓间充质干细胞并进行鉴定;②携带BMP-2和EGFP的慢病毒感染第3代BMSCs后,免疫荧光检测和RT-PCR及WB检测感染成功;③大鼠骨髓间充质干细胞感染慢病毒后进行骨向诱导,钙沉积明显增加;④BMSCs细胞黏附能力增强;⑤OPN、OCN、Col-Ⅰ及smad表达增加。结论:BMP-2通路增强了细胞黏附及相关成骨分化因子的表达,可明显促进大鼠骨髓间充质干细胞向成骨细胞定向分化,为BMSCs作为组织工程研究的种子细胞提供实验支持。
PURPOSE： To determine the effect of bone morphogenetic protein-2（BMP-2） on osteogenesis of bone mesenchymal stem cells（BMSCs） in rats.METHODS： The primary culture of rat BMSCs was succeeded and then was identified.The rat BMSCs were infected with the recombinant lentivirus with BMP-2（Lenti-BMP-2）.The osteogenic effect of BMP-2 was observed.In addition,adhesive ability of BMP-2-BMSCs was detected through adhesion assay and expression of osteogenic factors OPN,OCN,Col-Ⅰ and smad was observed by RT-PCR and Western blot.SPSS11.0 software package was used for analysis.RESULTS： Rat BMSCs were cultured and identified successfully.The osteogenic effect of BMSCs was improved by BMP-2.Lenti-BMP-2 BMSCs adhesive potential enhanced and osteogenic factors were up-regulated.CONCLUSIONS： BMP-2 may facilitate the osteogenic effect of the rat BMSCs and would provide favourable cell resource for tissue engineering.Supported by Natural Science Foundation of Shanghai Municipality（10JC1402600）.
出处
《上海口腔医学》
CAS
CSCD
2011年第4期352-357,共6页
Shanghai Journal of Stomatology
基金
上海市自然科学基金(10JC1402600)
复旦大学附属中山医院青年基金(337)
