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| mIL-18腺病毒载体构建及在大鼠骨髓间充质干细胞中的表达 | |
| 引用本文： | 许刚,郭燕舞,徐如祥,姜晓丹. mIL-18腺病毒载体构建及在大鼠骨髓间充质干细胞中的表达[J]. 西部医学, 2012, 24(6): 1067-1070 |
| 作者姓名： | 许刚 郭燕舞 徐如祥 姜晓丹 |
| 作者单位： | 1. 西安交通大学医学院第二附属医院神经外科,陕西西安,710004 2. 南方医科大学珠江医院神经外科,广东广州,510280 3. 北京军区总院神经外科,北京,100027 |
| 摘 要： | 目的构建表达mIL-18基因的腺病毒载体,并转染大鼠骨髓间充质干细胞(rBMSCs),观察其在rBMSCs中的表达,为脑胶质瘤的基因治疗实验研究奠定基础。方法 mIL-18基因亚克隆于穿梭质粒pAdtrackCMV上,构建的pAdtrackCMV-mIL-18线性化后转化已含有腺病毒骨架质粒pAdEasy-1的大肠杆菌BJ5183感受态细胞,挑选同源重组质粒,线性化后HEK293细胞包装,CsCI纯化。体外分离培养大鼠骨髓间充质干细胞,流式细胞仪检测细胞免疫表型;然后转染rBMSCs,通过荧光显微镜、RT-PCR、ELISA等方法检测mIL-18表达。结果成功构建了绿色荧光蛋白标记的腺病毒Ad-mIL-18,并在rBMSCs中高效表达。结论 pAdEasy-1是基因转染的高效系统,腺病毒Ad-mIL-18转染的rBMSCs可以作为脑肿瘤基因治疗研究的种子细胞。 |
| 关 键 词： | 白介素-18 腺病毒载体 骨髓间充质干细胞 |
Construction of adenovirus vectors contained mIL-18 and expression in rat mesenchymal stem cells |
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| Affiliation: | XU Gang1,GUO Yan-wu2,XU Ru-xiang3,et al(1.Department of Neurosurgery,The Second Affiliated Hospital,College of Medicine of Xi’an Jiaotong University,Xi’an Jiaotong University,Xi’an710004,China;2.Department of Neurosurgery,Zhujiang Hospital,South Medical University,Guangzhou510282,China;3.Department of Neurosurgery,Beijin Military District General Hospital,Beijin100027,China) |
| Abstract: | Objective To construct adenovirus vectors contained mIL-18and observation the expression in rBMSCs and explore experiment study for gene therapy of glioma.Methods mIL-18gene was subcloned into the shuttle plasmid pAdtrackCMV,then linearized the vector and convert competent cell BJ5183which contain the adenovirus frame plasmid pAdEasy-1.The homologous recombination plasmid was packaged by HEK293cells and purified by CsCI.The cell immunophenotype was detected by flow cytometry.The expression of mIL-18was measured by fluorescence microscope,RT-PCR and ELISA.Results The adenovirus vector Ad-mIL-18was successfully constructed,which was labeled by green fluorescent protein(GFP)and effectively expressed in rBMSCs.Conclusion pAdEasy-1is an effective gene transfection system.rBMSCs transfected with Ad-mIL-18can serve as the seeds cells for the gene therapy of glioma. |
| Keywords: | Interleukin-18 Adenovirus vector Bone marrow mesenchymal stem cell |
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