江苏科技信息
江囌科技信息
강소과기신식
JIANGSU SCIENCE & TECHNOLOGY INFORMATION
2015年
15期
65-70
,共6页
绿色荧光蛋白%慢病毒%骨髓间充质干细胞
綠色熒光蛋白%慢病毒%骨髓間充質榦細胞
록색형광단백%만병독%골수간충질간세포
green fluorescent protein%lentivirus%bone marrow mesenchymal stem cells
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。
關于組織工程骨構建領域種子細胞標記一直是研究熱點之一，構建GFP－重組慢病毒載體感染成骨方嚮誘導骨髓間充質榦細胞，具有重要的研究價值。文章採用RT－PCR方法檢測成骨基因骨橋蛋白和堿性燐痠酶、骨鈣素、堿性成纖維細胞生長因子和collgen-Ⅰ錶達，構建GFP－重組慢病毒載體，經基因測序和熒光倒置顯微鏡檢測其錶達。RT－PCR檢測結果顯示，骨髓間充質榦細胞成骨方嚮誘導培養成功，階段性錶達成骨基因骨橋蛋白和堿性燐痠酶、骨鈣素、堿性成纖維細胞生長因子和collgen-Ⅰ。基因測序結果證實GFP－重組慢病毒載體成功構建，熒光倒置顯微鏡檢測成功轉染成骨方嚮誘導培養骨髓間充質榦細胞，併在該細胞中錶達。文章最後得齣結論：GFP－重組慢病毒載體成功轉染成骨方嚮誘導的骨髓間充質榦細胞，GFP標記的骨髓間充質榦細胞具備良好的成骨功能，用于組織工程骨構建。
관우조직공정골구건영역충자세포표기일직시연구열점지일，구건GFP－중조만병독재체감염성골방향유도골수간충질간세포，구유중요적연구개치。문장채용RT－PCR방법검측성골기인골교단백화감성린산매、골개소、감성성섬유세포생장인자화collgen-Ⅰ표체，구건GFP－중조만병독재체，경기인측서화형광도치현미경검측기표체。RT－PCR검측결과현시，골수간충질간세포성골방향유도배양성공，계단성표체성골기인골교단백화감성린산매、골개소、감성성섬유세포생장인자화collgen-Ⅰ。기인측서결과증실GFP－중조만병독재체성공구건，형광도치현미경검측성공전염성골방향유도배양골수간충질간세포，병재해세포중표체。문장최후득출결론：GFP－중조만병독재체성공전염성골방향유도적골수간충질간세포，GFP표기적골수간충질간세포구비량호적성골공능，용우조직공정골구건。
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.
