發(fā)布時間：2018-07-01 13:14

  本文選題：骨形態(tài)發(fā)生蛋白 + 慢病毒載體　； 參考：《口腔醫(yī)學研究》2015年01期

【摘要】：目的:構(gòu)建骨形態(tài)發(fā)生蛋白4(BMP4)慢病毒載體,檢測其對鼠骨髓間充質(zhì)干細胞成牙功能的影響,以期為組織工程牙篩選種子細胞。方法:以成熟人胎盤組織為材料來源,克隆人BMP4基因的全長cDNA,轉(zhuǎn)克隆至plenti6/v5-D-TOPO表達載體,構(gòu)建出BMP4-plenti6/v5-D-TOPO重組慢病毒表達載體;轉(zhuǎn)染SD大鼠骨髓間充質(zhì)干細胞,用MTT法檢測轉(zhuǎn)染前后細胞的增殖活性,實時熒光定量PCR檢測Ⅰ型膠原蛋白、成釉蛋白、牙本質(zhì)基質(zhì)蛋白1、同源異型盒基因1成牙相關基因的mRNA水平相對表達量的變化。結(jié)果:BMP4基因轉(zhuǎn)染后,細胞體外增殖活性增強,成釉蛋白、牙本質(zhì)基質(zhì)蛋白1、同源異型盒基因1mRNA水平表達量增多,差異有統(tǒng)計學意義(P0.05),Ⅰ型膠原蛋白mRNA水平差異無顯著性意義。結(jié)論:BMP4可提高骨髓間充質(zhì)干細胞體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，

本文編號：2087903