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目的比较微小RNA(miRNAs)在人牙髓干细胞(dental pulp stem cells,DPSCs)及非DPSCs中的表达差异,探讨其在维持DPSCs干性状态中的作用。方法本研究于2013年1—10月在福建医科大学附属口腔医院完成。原代培养人牙髓细胞,利用结合了STRO-1特异性抗体的免疫磁珠分选获得DPSCs,并进行成牙本质样细胞的诱导分化,检测碱性磷酸酶(ALP)、骨钙素(OC)值以及进行von Kossa染色,鉴定其分化能力。采用miRNA基因芯片技术,检测DPSCs和非DPSCs中miRNAs的表达,筛选出差异表达的miRNAs。结果与非DPSCs相比,DPSCs中表达上调超过2倍的miRNAs有11个,表达下调超过2倍的miRNAs有3个。结论 miRNAs表达谱的变化可能与DPSCs干性状态的维持相关。
Objective To compare the expression of miRNAs in human dental pulp stem cells (DPSCs) and non-DPSCs and to explore the role of miRNAs in maintaining the dry state of DPSCs. Methods The study was performed in Stomatological Hospital Affiliated to Fujian Medical University from January to October in 2013. Human dental pulp cells were cultured in primary culture. DPSCs were sorted by immunomagnetic beads combined with antibodies specific to STRO-1. Differentiation of odontoblast-like cells was performed. Alkaline phosphatase (ALP), osteocalcin OC) values and von Kossa staining to identify their ability to differentiate. MiRNA microarray was used to detect the expression of miRNAs in DPSCs and non-DPSCs, and the differentially expressed miRNAs were screened out. Results Compared with non-DPSCs, there were 11 miRNAs that were up-regulated more than 2-fold in DPSCs and 3 miRNAs that were more than 2-fold down-regulated. Conclusion The changes of miRNAs expression may be related to the maintenance of the dry state of DPSCs.