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论文摘要

过表达骨形态发生蛋白-4对小鼠诱导性多能干细胞生物学活性的影响

Effect of bone morphogenetic protein-4 overexpression on the biological activity of mouse induced pluripotent stem cells

作者:裘今, 张慧宇, 刘丽, 谭颖徽

Author:Jin Qiu, Huiyu Zhang, Li Liu, Yinghui Tan

收稿日期:2017-09-16          年卷(期)页码:2018,36(2):190-190-193

期刊名称:华西口腔医学杂志

Journal Name:West China Journal of Stomatology

关键字:骨形态发生蛋白-4,诱导性多能干细胞,牙向分化,增殖,

Key words:bone morphogenetic protein-4,induced pluripotent stem cells,odontogenic differentiation,proliferation,

基金项目:国家自然科学基金(31400841)

中文摘要

目的 构建过表达骨形态发生蛋白-4(BMP4)基因的慢病毒载体,探讨其过表达后对小鼠诱导性多能干细胞(iPS)生物学活性的影响。方法 采用慢病毒转染建立稳定过表达BMP4的iPS细胞株(BMP4过表达组),未作任何转染的细胞为空白组,转染慢病毒阴性对照组的细胞为空载体组。CCK8检测各组细胞增殖活性,碱性磷酸酶(ALP)活性检测细胞分化程度,荧光定量聚合酶链反应检测BMP4、成釉细胞蛋白(AMBN)、细胞角蛋白(CK)14、牙本质涎磷蛋白(DSPP)、骨唾液酸蛋白(BSP)及骨细胞特异转录因子Runx2的mRNA表达。结果 与空白组及空载体组相比,BMP4过表达组的细胞增殖活性及ALP活性升高(P<0.05),Runx2、AMBN、CK14、DSPP、BSP mRNA的表达增加(P<0.05)。结论 BMP4基因能够促进iPS的牙向分化。

英文摘要

ObjectiveThis study aimed to construct the expression of bone morphogenetic protein-4 (BMP4) lentiviral vector gene and explore its influence on the biological activity of mouse induced pluripotent stem (iPS) cells.MethodsiPS cell lines stably overexpressing BMP4 were constructed by lentivirus transfection (BMP4-overexpressing group). Cells without transfection served as the blank group, and cells with only vector transfection served as the empty-vector group. Cell prolifera-tion was detected by CCK8, and the expression levels of ameloblastin (AMBN), cytokeratin (CK) 14, dentin sialophospho-protein (DSPP), bone sialoprotein (BSP), and Runx2 mRNA were detected by quantitative polymerase chain reaction. Alkaline phosphatase (ALP) activity was used to detect the degree of cell differentiation.ResultsCompared with blank and empty-vector groups, proliferation activity and ALP activity of BMP4-overexpressing group obvious increased (P<0 .05), bmp4, ambn, ck14, dspp, bsp, runx2 mrna expression also increased (P<0 .05).ConclusionBMP4 can significantly promote the odontogenic differentiation of iPS.

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