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目的探讨人牙周膜干细胞(periodontal ligament stem cells,PDLSCs)成骨分化过程中miRNA表达谱的变化,以期为揭示PDLSCs成骨分化的分子机制提供基础。方法本研究于2012年1—12月在同济大学口腔医学院口腔生物医学及转化医学实验室、复旦大学公共实验平台完成。原代培养人PDLSCs,经流式细胞仪检测鉴定其干细胞特性(表达干细胞表面标志物STRO-1);体外诱导其往成骨方向分化,经茜素红S及碱性磷酸酶显色试剂盒染色检测其成骨特性。再采用表达谱基因芯片,分析成骨诱导前后PDLSCs的miRNA表达,筛选出分化前后差异表达的miRNA。结果与未经成骨诱导的细胞相比,PDLSCs在成骨分化14d后,有11个miRNA的表达上调,13个miRNA的表达下调。结论 PDLSCs具有成骨潜能,其成骨分化的机制可能与miRNA表达的改变有关。
Objective To investigate the changes of miRNA expression profiles in the process of osteogenic differentiation of human periodontal ligament stem cells (PDLSCs), in order to provide the basis for revealing the molecular mechanism of PDLSCs osteogenic differentiation. Methods The study was performed in the oral biomedical and translational medicine laboratory of Tongji University from January to December 2012 and the public experiment platform of Fudan University. Primary cultured human PDLSCs were identified by flow cytometry to identify the characteristics of stem cells (express stem cell surface marker STRO-1); induced in vitro into osteoblasts and differentiated into osteogenic cells by alizarin red S and alkaline phosphatase assay kit Staining detected its osteogenic properties. Then using the expression profile gene chip, miRNA expression of PDLSCs before and after osteogenesis was analyzed, and miRNAs differentially expressed before and after differentiation were screened. Results Compared with the non-osteogenic cells, 11 miRNAs were up-regulated and 13 miRNAs were down-regulated after 14d osteogenic differentiation. Conclusion PDLSCs have osteogenic potential. The mechanism of osteogenic differentiation may be related to the changes of miRNA expression.