骨形态发生蛋白4慢病毒载体构建及对鼠骨髓间充质干细胞成牙功能影响

发布时间：2018-07-01 13:14

  本文选题：骨形态发生蛋白 + 慢病毒载体　； 参考：《口腔医学研究》2015年01期

【摘要】：目的:构建骨形态发生蛋白4(BMP4)慢病毒载体,检测其对鼠骨髓间充质干细胞成牙功能的影响,以期为组织工程牙筛选种子细胞。方法:以成熟人胎盘组织为材料来源,克隆人BMP4基因的全长cDNA,转克隆至plenti6/v5-D-TOPO表达载体,构建出BMP4-plenti6/v5-D-TOPO重组慢病毒表达载体;转染SD大鼠骨髓间充质干细胞,用MTT法检测转染前后细胞的增殖活性,实时荧光定量PCR检测Ⅰ型胶原蛋白、成釉蛋白、牙本质基质蛋白1、同源异型盒基因1成牙相关基因的mRNA水平相对表达量的变化。结果:BMP4基因转染后,细胞体外增殖活性增强,成釉蛋白、牙本质基质蛋白1、同源异型盒基因1mRNA水平表达量增多,差异有统计学意义(P0.05),Ⅰ型胶原蛋白mRNA水平差异无显著性意义。结论:BMP4可提高骨髓间充质干细胞体外增殖活性和牙向分化能力。
[Abstract]:Aim: to construct bone morphogenetic protein 4 (BMP4) lentivirus vector and to detect the effect of BMP4 on dental function of bone marrow mesenchymal stem cells (MSCs) in order to screen seed cells for tissue engineering teeth. Methods: the full-length cDNAof human BMP4 gene was cloned from mature human placental tissue and cloned into plenti6 / v5-D-Topo expression vector. The recombinant lentivirus expression vector BMP4-plenti6 / v5-D-TOPO was constructed and transfected into SD rat bone marrow mesenchymal stem cells. The proliferative activity of the cells before and after transfection was detected by MTT assay. The mRNA expression of type I collagen protein, amelogenin, dentin matrix protein 1 and homologous box gene 1 was detected by real-time quantitative PCR. Results after transfection of BMP4 gene, the proliferative activity of BMP4 cells was enhanced, and the expression of amelogenin, dentin matrix protein 1 and homologous box gene 1 mRNA was increased (P0.05), but there was no significant difference in type 鈪，

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