| 腺病毒介导人VEGF_(165)和ANG-1双基因转染大鼠BMSCs及对其增殖的影响 |
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| 引用本文： | 张金康,孟国林,刘建,胡蕴玉,袁志,段春光,白峰,白建萍,李丹.腺病毒介导人VEGF_(165)和ANG-1双基因转染大鼠BMSCs及对其增殖的影响[J].中国矫形外科杂志,2011(11). |
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| 作者姓名： | 张金康 孟国林 刘建 胡蕴玉 袁志 段春光 白峰 白建萍 李丹 |
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| 作者单位： | 第四军医大学西京骨科医院全军骨科研究所; |
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| 基金项目： | 国家自然科学基金(编号:30672142) |
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| 摘 要： | 目的]探讨携带人血管内皮细胞生长因子165(VEGF165)和血管生成素-1(ANG-1)双基因的腺病毒表达载体pAd-VIA转染大鼠骨髓基质干细胞(BMSCs)的体外表达及对BMSCs增殖的影响。方法]将本室构建的编码人VEGF165和ANG-1双基因的腺病毒质粒pAd-VIA在QBI-293A细胞内进行包装和扩增,体外转染大鼠BMSCs,通过绿色荧光蛋白(GFP)表达、Western-blotting、酶联免疫吸附分析(ELISA)方法检测外源基因的表达,利用MTT法检测MOI(multiplicity of infection)=50、100、200、400 pfu/cell不同浓度腺病毒转染后对BMSCs增殖活性的影响。结果]重组腺病毒质粒pAd-VIA在QBI-293A细胞内成功包装和扩增,体外转染BMSCs后,有大量的GFP表达,Western-blotting检测显示,转染组与VEGF165和ANG-1抗体结合,在45 KD和14.4 KD左右出现印迹条带;ELISA结果显示:转染组第1、2、3 d,上清中的VEGF165浓度和ANG-1浓度持续增加,未转染组均未检测到外源基因的表达,差异...
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| 关 键 词： | 血管内皮细胞生长因子165 血管生成素-1 骨髓基质干细胞 增殖 |
Effect of adenoviruses-mediated human angiopoietin-1 and VEGF_(165) gene over-expression on ex vivo growth capability of rat bone marrow stromal cells |
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| ZHANG Jin-kang,MENG Guo-lin,LIU Jian,et al..Effect of adenoviruses-mediated human angiopoietin-1 and VEGF_(165) gene over-expression on ex vivo growth capability of rat bone marrow stromal cells[J].The Orthopedic Journal of China,2011(11). |
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| Authors: | ZHANG Jin-kang MENG Guo-lin LIU Jian |
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| Institution: | ZHANG Jin-kang,MENG Guo-lin,LIU Jian,et al.Institute of Orthopaedics and Traumatology,Xijing Hospital,the Fourth Military Medical University,Xi'an 710032,China |
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| Abstract: | Objective]The purpose of this study was to investigate the effect of adenoviruses-mediated human angiopoietin-1 and VEGF165 expression on ex vivo growth capability of rat bone marrow stromal cells. Methods]The recombinant pAd-VIA plasmid constructed by our laboratory was packaged and amplified in QBI-293A cells.The pAd-VIA recombined adenovirus was obtained.Rat bone marrow stromal cells(BMSCs) were infected with pAd-VIA recombined adenovirus at about 80% confluency.The expression of the gene of interests ... |
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| Keywords: | VEGF165 angiopoietin-1 bone marrow stromal cells proliferation |
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