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Differential expression of drug resistance genes in CD146 positive dental pulp derived stem cells and CD146 negative fibroblasts

机译:CD146阳性牙髓衍生干细胞和CD146阴性成纤维细胞中耐药基因的差异表达

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Introduction The stem cell portion of the dental pulp derived cultures (DPSCs) showed a higher resistance to cytotoxic effect of restorative dental materials compared to pulpal fibroblasts (DPFs). Here, we aimed to compare the expression of some drug resistant genes between these cells. Methods and materials To separate DPSCs from DPFs, we used magnetic cell sorting technique based on CD146 expression. To assess the stem cell properties, the positive and negative portions underwent colony forming assays and were induced to be differentiated into the adipocytes, osteoblasts, hepatocytes, and neural cells. Cell surface antigen panels were checked using immune fluorescence and flow‐cytometry techniques. The mRNA expression of 14 ABC transporters including ABCA2, ABCB1, ABCB11, ABCC1, ABCC2, ABCC3, ABCC4, ABCC5‐2, ABCC5‐4,ABCC5‐13, ABCC6, ABCC10, ABCC11, and ABCG2 genes was assessed, using quantitative RT‐PCR technique. Results Only the CD146 positive portion could be differentiated into the desired fates, and they formed higher colonies (16.7?±?3.32 vs. 1.7?±?1.67, p??.001). The cell surface antigen panels were the same, except for CD146 and STRO‐1 markers which were expressed only in the positive portion. Among the ABC transporter genes studied, the positive portion showed a higher expression (approximately two‐fold) of ABCA2, ABCC5‐13, and ABCC5‐2 genes. Conclusion Dental pulp stem cells which can be separated from dental pulp fibroblasts based on CD146 expression, express higher levels of some drug resistance genes which probably accounts for their features of more resistance to cytotoxic effects of some dental materials. This needs to be more validated in future.
机译:引入牙髓来源的培养物(牙髓干细胞)的干细胞部分表明相比于牙髓成纤维细胞(DPF中)复原牙用材料的细胞毒性作用更高的电阻。在这里,我们的目的是比较这些细胞之间的一些耐药基因的表达。方法和材料从DPF中分离牙髓干细胞,我们使用了基于CD146表达磁性细胞分选技术。为了评估干细胞特性,正和负部分后行集落形成测定法和被诱导分化成脂肪细胞,成骨细胞,肝细胞和神经细胞。使用免疫荧光细胞表面抗原的板检查和流式细胞术技术。的14个ABC转运包括ABCA2,ABCB1,ABCB11,ABCC1,ABCC2,ABCC3,ABCC4,ABCC5-2,ABCC5-4,ABCC5-13,ABCC6,ABCC10,ABCC11,和ABCG2基因的mRNA表达进行评价,使用定量RT- PCR技术。结果只有CD146阳性部分可以分化成所期望的命运,并且它们形成更高的菌落(16.7?±3.32对1.7〜±1.67,P <??001)。细胞表面抗原的面板是相同的,除了CD146并且其表达仅在正部分STRO-1标记。在所研究的ABC转运蛋白基因,正部分显示出更高的表达ABCA2,ABCC5-13,和ABCC5-2基因(大约两倍)。它可以从基于CD146表达牙髓成纤维细胞中分离结论牙髓干细胞,表达更高水平的一些这可能解释了它们对一些牙科材料的细胞毒作用更电阻的特征药物抗性基因。这需要在未来更多的验证。

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