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| 携带肾上腺髓质素基因腺病毒体外转染骨髓间充质干细胞及其蛋白的表达 | |
| 引用本文： | 林茂海,苏津自,蔡文钦,郑礼裕,许昌声,王华军.携带肾上腺髓质素基因腺病毒体外转染骨髓间充质干细胞及其蛋白的表达[J].中国组织工程研究与临床康复,2011,15(23). |
| 作者姓名： | 林茂海 苏津自 蔡文钦 郑礼裕 许昌声 王华军 |
| 作者单位： | 1. 莆田市第一医院心内科,福建省莆田市,351100 2. 福建医科大学附属第一医院心内科,福建省福州市,350005 3. 福建医科大学附属第一医院高血压研究所,福建省福州市,350005 |
| 基金项目： | 福建医科大学附属第一医院"院学科带头人"科研基金 |
| 摘 要： | 背景:肾上腺髓质素具有促进骨髓间充质干细胞增殖、生长及减少凋亡等作用.目的:观察携带肾上腺髓质素基因的腺病毒体外转染骨髓间充质干细胞及其蛋白的表达.方法:培养293A细胞,并扩增携带肾上腺髓质素基因的腺病毒(Ad-ADM),实验分为对照组、空载体组和Ad-ADM转染组,3组分别于细胞长满至50%~60%时加入等量的无血清L-DMEM、Ad-lacZ和Ad-ADM,噬斑法检测扩增后腺病毒的滴度;贴壁法体外分离、扩增培养大鼠骨髓间充质干细胞;Ad-GFP体外感染骨髓间充质干细胞后检测转染效率;RT-PCR检测Ad-ADM转染后肾上腺髓质素mRNA的表达.结果与结论:病毒扩增后噬斑试验测定Ad-ADM的病毒滴度为1.1×109 pfu/mL.Ad-ADM感染骨髓间充质干细胞后,肾上腺髓质素mRNA的表达明显增加,对照组、空载体组未见ADM mRNA表达;量化结果表明其表达量从第1天开始逐渐增加,第7天达高峰,在11 d时仍可检测其表达.提示腺病毒对骨髓间充质干细胞有较高的转染效率.感染Ad-ADM后,骨髓间充质干细胞可有效表达肾上腺髓质素. |
| 关 键 词： | 肾上腺髓质素 293细胞 噬斑试验 骨髓间充质干细胞 腺病毒载体 |
Adenovirus carrying adrenomedullin gene for transfection of bone marrow mesenchymal stem cells in vitro and their protein expression |
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| Lin Mao-hai,Su Jin-zi,Cai Wen-qin,Zheng Li-yu,Xu Chang-sheng,Wang Hua-jun.Adenovirus carrying adrenomedullin gene for transfection of bone marrow mesenchymal stem cells in vitro and their protein expression[J].Journal of Clinical Rehabilitative Tissue Engineering Research,2011,15(23). | |
| Authors: | Lin Mao-hai Su Jin-zi Cai Wen-qin Zheng Li-yu Xu Chang-sheng Wang Hua-jun |
| Institution: | Lin Mao-hai1,Su Jin-zi2,Cai Wen-qin2,Zheng Li-yu2,Xu Chang-sheng3,Wang Hua-jun3 1Department of Cardiology,The First Hospital of Putian City,Putian 351100,Fujian Province,China,2Department of Cardiology,The First Affiliated Hospital,Fujian Medical University,Fuzhou 350005,3Fujian Institute of Hypertension Research,The First Affiliated Hospital of Fujian Medical University |
| Abstract: | BACKGROUND:Adrenomedullin is one of potential growth factor that can promote cellular differentiation,growth and reduce apoptosis.OBJECTIVE:To investigate the possibility of bone marrow mesenchymal stem cells transfected by adenovirus vector carrying adrenomedullin(Ad-ADM) ,and to determine the protein expression of adrenomedullin following transfection.METHODS:The 293A cells were cultivated and the Ad-ADM was amplified.The experiment was divided into three groups:control,blank vector and Ad-ADM transfectio... |
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