人牙周膜干细胞成骨分化前后miRNA表达谱差异分析  

Variation analysis of miRNA expression during osteogenic differentiation of human periodontal ligament stem cells

作　　者：章燕 甄蕾[3] 郭姜莉 

机构地区：[1]同济大学附属口腔医院口腔颌面外科 [2]同济大学口腔医学院口腔生物医学及转化医学实验室,上海200072 [3]上海市口腔病防治院口腔内科,上海200001

出　　处：《中国实用口腔科杂志》2013年第3期148-152,共5页Chinese Journal of Practical Stomatology

基　　金：上海市卫生局青年基金项目(20124y034);上海市青年医师培养资助计划(人才-2012-04-03)

摘　　要：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 analyze the differential expression of miRNAs in periodontal ligament stem cells （PDLSCs） and osteo-differentiated PDLSCs. Methods PDLSCs were isolated and cultivated. PDLSCs of passage 2 was plated at density of 5×10^3 / cm^2. At 80% confluence, the medium was changed to an inducing medium. After 14 days of induction, the results were evaluated by Alizarin red S staining, ALP activity test and miRNA microarrays. Results PDLSCs were cultivated and differentiated in vitro with active function, which were manifested by Alizarin red S staining and ALP activity test. miRNA microarrays revealed that 13 miRNAs were downregulated in osteo-differentiated PDLSCs while 11 miRNAs were upregulated. Conclusions PDLSCs have the potential to differentiate into osteoblasts, miRNA may play an important role in the osteogenic differentiation of periodontal ligament stem cells.

关 键 词：牙周膜干细胞 成骨分化 MIRNA 

分 类 号：R78[医药卫生—口腔医学]