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牙胚细胞与骨形态发生蛋白4转染的骨髓间充质干细胞相互诱导的体外实验研究

Study on the Mutual Induction between Tooth Germ Cells and Bone Marrow Mesenchymal Stem Cells Transfected by Bone Morphogenetic Protein4 Gene in Vitro
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摘要 目的:探讨牙胚细胞和骨形态发生蛋白4(BMP4)转染的骨髓间充质干细胞(BMSCs)相互间的成牙诱导作用。方法:构建骨形态发生蛋白4慢病毒载体,转染SD大鼠骨髓间充质干细胞,并将骨髓间充质干细胞和BMP4转染的骨髓间充质干细胞分别与SD大鼠牙胚细胞按1∶1比例混合培养,同时将细胞分为5组,BMSCs组、BMP4/BMSCs组、牙胚细胞组、BMSCs/牙胚细胞组(混合组1)、BMP4/BMSCs/牙胚细胞组(混合组2),分别采用实时荧光定量PCR和western-blotting检测五组细胞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mRNA水平和蛋白水平相对表达量的变化。结果:与混合组1相比,混合组2Ⅰ型胶原蛋白、成釉蛋白、牙本质基质蛋白1、同源异型盒基因1mRNA水平和蛋白水平表达量增多,差异有统计学意义(P<0.05)。结论:牙胚细胞与BMP4转染的骨髓间充质干细胞共培养,促进了成牙相关基因的表达,可作为组织工程牙的备选种子细胞。 Objective: To study the mutual induction between tooth germ cells and bone marrow mesenchymal stem cells with bone morphogenetic protein4 gene in vitro. Methods: Lentiviral vectors carrying BMP4 were con- structed to transfect rat bone marrow mesenchymal stem cells(BMSCs) and BMSCs tansfected by BMP4 were co-- cultured with tooth germ cells (1:1)respectively . The cells were divided into five groups: BMSCs group, BMP4/ BMSCs group, tooth germ cells group, BMSCs tooth germ cells group (the mixed groupl), BMP4/BMSCs tooth germ cells group (the mixed group2) . The five groups were extracted using real--time quantitative PCR and west- ern blotting to detect the mRNA and protein levels of collagen I, enamel protein, dentin matrix protein 1, and ho- meobox gene 1. Results: In the mixed group2, mRNA and protein levels of collagen I, enamel protein,dentin matrix protein 1 and homeobox gene 1 were higher, and the difference was statistically significant (P〈0.05). Conclusion: The co--culture of tooth germ cells and BMSCs tansfected by BMP4 can increase the expressions of odontogenic genes, and they are suitable as seed cells of tissue--engineered tooth.
出处 《口腔医学研究》 CAS CSCD 北大核心 2015年第3期237-241,共5页 Journal of Oral Science Research
基金 新疆维吾尔自治区科技厅科技支疆项目(编号:201291173) 新疆维吾尔自治区科技厅自然科学基金(编号:20142110037) 新医大一附院组织工程专项(编号:2012ZZGC01)
关键词 牙胚细胞 骨髓间充质干细胞 骨形态发生蛋白4 实时荧光定量PCR western-blotting Tooth germ cells Bone mesenchymal stem cells Bone morphogenetic protein4 Real--time fluo-rescent quantitative PCR Western--blotting
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参考文献12

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