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江苏科技信息 ›› 2015, Vol. 32 ›› Issue (15): 61-64.doi: 10.3969/j.issn.1004-7530.2015.15.022

• 论文 • 上一篇    下一篇

绿色荧光蛋白-重组慢病毒载体转染骨向诱导的骨髓间充质干细胞及鉴定

梁学萍, 张蕾, 胡杨   

  1. 新疆医科大学第一附属医院牙体牙髓科,新疆乌鲁木齐,830054;新疆医科大学第一附属医院口腔修复科,新疆乌鲁木齐,830054
  • 出版日期:2015-05-25 发布日期:2015-05-25
  • 基金资助:
    新疆维吾尔自治区科技厅自然科学基金%项目编号2013211A082。新疆维吾尔自治区科技厅自然科学基金%项目编号2014211C037。

Bone Marrow Mesenchymal Stem Cells Transfected by GFP Recombinant Lentivirus and Its Identification

Liang Xueping, Zhang Lei, Hu Yang   

  • Online:2015-05-25 Published:2015-05-25

摘要： 关于组织工程骨构建领域种子细胞标记一直是研究热点之一，构建GFP－重组慢病毒载体感染成骨方向诱导骨髓间充质干细胞，具有重要的研究价值。文章采用RT－PCR方法检测成骨基因骨桥蛋白和碱性磷酸酶、骨钙素、碱性成纤维细胞生长因子和collgen-Ⅰ表达，构建GFP－重组慢病毒载体，经基因测序和荧光倒置显微镜检测其表达。RT－PCR检测结果显示，骨髓间充质干细胞成骨方向诱导培养成功，阶段性表达成骨基因骨桥蛋白和碱性磷酸酶、骨钙素、碱性成纤维细胞生长因子和collgen-Ⅰ。基因测序结果证实GFP－重组慢病毒载体成功构建，荧光倒置显微镜检测成功转染成骨方向诱导培养骨髓间充质干细胞，并在该细胞中表达。文章最后得出结论：GFP－重组慢病毒载体成功转染成骨方向诱导的骨髓间充质干细胞，GFP标记的骨髓间充质干细胞具备良好的成骨功能，用于组织工程骨构建。

关键词: 绿色荧光蛋白, 慢病毒, 骨髓间充质干细胞

Abstract: The method of labeled seed cells is one of hot research of the construction of tissue engineering bone field, construction of GFP recombinant lentiviral vector infected BMSCs induced into osteoblasts, and it has important research value.BMSCs were cultured by bone induced medium, expression of bone gene of alkaline phosphatase, osteocalcin and osteopontin, basic fibroblast growth factor and collgen-I detected by RT- PCR method. The construction of GFP recombinant lentiviral vector, gene sequencing and inverted fluorescence microscope detected its expression.Results of RT-PCR showed BMSCs successfully induced into osteoblasts, osteogenic gene of osteopontin and osteocalcin, alkaline phosphatase, basic fibroblast growth factor and collgen-I expressed at stage.Gene sequencing results confirmed that GFP recombinant lentiviral vector was successfully constructed, inverted fluorescence microscope detection, BMSCs induced into osteoblasts were successfully transfected by GFP recombinant lentiviral vector ,and the GFP expressed in the BMSCs.The GFP recombinant lentiviral vector successfully transfected BMSCs induced into osteoblasts ,GFP labeled BMSCs are good osteogenic function for the construction of tissue engineered bone.