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| 慢病毒介导的稳定表达人GM-CSF、IL-4的EA.hy926细胞株的建立 | |
| 引用本文： | 石晓月,贺敏,任鹏,宋丽雅,乔志新,杨晓琮,程宏,冯秋月,张公庆,王璇琳,宫宁,王字玲,李伟静,于群.慢病毒介导的稳定表达人GM-CSF、IL-4的EA.hy926细胞株的建立[J].生物技术通讯,2011,22(4):480-483. |
| 作者姓名： | 石晓月 贺敏 任鹏 宋丽雅 乔志新 杨晓琮 程宏 冯秋月 张公庆 王璇琳 宫宁 王字玲 李伟静 于群 |
| 作者单位： | 1. 军事医学科学院野战输血研究所,北京,100850 2. 北京工商大学,理学院,北京,100048 3. 解放军316医院,北京,100093 |
| 基金项目： | 国家传染病重大专项，北京市病毒性肝炎防治技术支撑体系建设重大专项 |
| 摘 要： | 目的:获得能够稳定表达人粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素4(IL-4)的内皮细胞株。方法:构建重组慢病毒表达载体,在293FT细胞中进行病毒的包装,用获得的高滴度慢病毒感染EA.hy926细胞株,建立能够稳定、高效表达hGM-CSF、hIL-4的EA.hy926细胞株。结果:EA.hy926感染效率达90%以上,长期传代后阳性率仍维持在90%以上;实时定量PCR和ELISA方法证实感染后细胞hGM-CSF、hIL-4在mRNA水平的表达分别是未感染细胞的25倍和9946倍,在蛋白水平的表达分别是未感染细胞的145倍和23倍。结论:构建的细胞系可以稳定表达细胞因子GM-CSF、IL-4。 |
| 关 键 词： | 树突状细胞 转染 白细胞介素4 粒细胞-巨噬细胞集落刺激因子 |
Construction and Identification of Cell Line to Coexpress GM-CSF and IL-4 Mediated by Lentiviral System |
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| SHI Xiao-Yue,HE Min,REN Peng,SONG Li-Ya,QIAO Zhi-Xin,YANG Xiao-Cong,CHENG Hong,FENG Qiu-Yue,ZHANG Gong-Qing,WANG Xuan-Lin,GONG Ning,WANG Zi-Ling,LI Wei-Jing,YU Qun.Construction and Identification of Cell Line to Coexpress GM-CSF and IL-4 Mediated by Lentiviral System[J].Letters in Biotechnology,2011,22(4):480-483. | |
| Authors: | SHI Xiao-Yue HE Min REN Peng SONG Li-Ya QIAO Zhi-Xin YANG Xiao-Cong CHENG Hong FENG Qiu-Yue ZHANG Gong-Qing WANG Xuan-Lin GONG Ning WANG Zi-Ling LI Wei-Jing YU Qun |
| Institution: | SHI Xiao-Yue1,HE Min1,REN Peng1,SONG Li-Ya2,QIAO Zhi-Xin1,YANG Xiao-Cong1,CHENG Hong1,FENG Qiu-Yue1,ZHANG Gong-Qing1,WANG Xuan-Lin1,GONG Ning3,WANG Zi-Ling1,LI Wei-Jing1,YU Qun1 1.Beijing Institute of Transfusion Medicine,Beijing 100850,2.School of Science,Beijing Technology and Business University,Beijing 100048,3.The 316th Hospital of PLA,Beijing 100093,China |
| Abstract: | Objective: To establish the EA.hy926 cell line expressing granulocyte-macrophage colony stimulating factor(GM-CSF) and interleukin 4(IL-4) stably mediated by lentiviral system.Methods: The recombinant lentiviral plasmid carrying human GM-CSF and IL-4 was transfected into 293FT cells with lentiviral packaging plasmids.Then the infectious lentiviral was obtained,which infected the EA.hy926 cells.The expression of GM-CSF and IL-4 was assayed by qPCR and ELISA.Results: The GM-CSF and IL-4 expression at mRNA lev... |
| Keywords: | dendritic cell transfection interleukin 4 granulocyte-macrophage colony stimulating factor |
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