| 重组pcDNA3.1-hBMP-2转染兔脂肪干细胞体外诱导的成骨分化 |
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| 引用本文： | 安荣泽,王兆杰,刘凡凡,齐新文,袁小洪,陈军平,赵俊延,胡小军,杨晋,赵豪.重组pcDNA3.1-hBMP-2转染兔脂肪干细胞体外诱导的成骨分化[J].中国组织工程研究与临床康复,2011,15(41):7601-7606. |
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| 作者姓名： | 安荣泽 王兆杰 刘凡凡 齐新文 袁小洪 陈军平 赵俊延 胡小军 杨晋 赵豪 |
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| 作者单位： | 遵义医学院第五附属(珠海)医院,广东省珠海市,519100 |
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| 基金项目： | 贵州省省长基金课题"组织工程软骨"资助，珠海市医学重点建设专科基金 |
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| 摘 要： | 背景:骨形态发生蛋白2具有很强的诱导干细胞成骨活性。目的:构建人骨形态发生蛋白2基因真核表达载体,探讨其转染脂肪干细胞后的成骨效果。方法:通过噬菌斑原位杂交筛选人混合细胞cDNA文库获得人骨形态发生蛋白2基因,与真核表达载体pcDNA3.1-连接,构建重组质粒pcDNA3.1-hBMP-2。利用脂质体LipofectamineTM2000分别介导人骨形态发生蛋白2基因、EGFP基因转染第4代脂肪干细胞,并经G418进行筛选。结果与结论:酶切鉴定及DNA测序结果证实重组质粒pcDNA3.1-hBMP-2构建成功。经计算脂质体介导的脂肪干细胞瞬时转染率为(18.0±0.42)%,并经过G418筛选后获得了稳定转染的细胞。细胞生长曲线表明转染后对脂肪干细胞生长、增殖无明显影响。ELISA检测发现人骨形态发生蛋白2组的人骨形态发生蛋白2因子表达量均高于EGFP组及未转染组,且能够稳定表达。经人骨形态发生蛋白2基因转染的脂肪干细胞的Ⅰ型胶原含量、碱性磷酸酶活性以及钙结节数目均比EGFP组及未转染组有明显升高。
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| 关 键 词： | 骨形态发生蛋白2 脂肪源性干细胞 脂质体 转染 骨组织工程 |
Osteogenic differentiation of rabbit adipose derived stem cells transfected with recombinant pcDNA3.1-hBMP-2 in vitro |
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| An Rong-ze,Wang Zhao-jie,Liu Fan-fan,Qi Xin-wen,Yuan Xiao-hong,Chen Jun-ping,Zhao Jun-yan,Hu Xiao-jun,Yang Jin,Zhao Hao.Osteogenic differentiation of rabbit adipose derived stem cells transfected with recombinant pcDNA3.1-hBMP-2 in vitro[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2011,15(41):7601-7606. |
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| Authors: | An Rong-ze Wang Zhao-jie Liu Fan-fan Qi Xin-wen Yuan Xiao-hong Chen Jun-ping Zhao Jun-yan Hu Xiao-jun Yang Jin Zhao Hao |
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| Institution: | An Rong-ze,Wang Zhao-jie,Liu Fan-fan,Qi Xin-wen,Yuan Xiao-hong,Chen Jun-ping,Zhao Jun-yan,Hu Xiao-jun,Yang Jin,Zhao Hao Orthopedic Department of the Fifth Affiliated Hospital of Zunyi Medical College,Zhuhai 519100,Guangdong Province,China |
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| Abstract: | BACKGROUND:Bone morphogenetic protein 2(BMP-2) shows strong potential to induce osteogenesis of stem cells.OBJECTIVE:To construct an eukaryotic expression vector of human BMP-2(hBMP-2) gene and then to investigate its effect in osteogenic differentiation of human adipose-derived stem cells(ADSCs).METHODS:hBMP-2 gene was obtained by screening human mixed cell cDNA library by plaque in situ hybridization and connected with the eukaryotic expression vector pcDNA3.1.The recombinant plasmid pcDNA3.1-hBMP-2 was i... |
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