| 腺病毒载体介导NGF基因转染大鼠肌源性干细胞的实验研究 |
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| 引用本文： | 方建雄,刘豪圣,刘天琦,张振辉,刘久敏,蒲小勇. 腺病毒载体介导NGF基因转染大鼠肌源性干细胞的实验研究[J]. 国际泌尿系统杂志, 2021, 41(1): 109-113 |
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| 作者姓名： | 方建雄 刘豪圣 刘天琦 张振辉 刘久敏 蒲小勇 |
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| 作者单位： | 广东省人民医院（广东省医学科学院）泌尿外科，广州 510055 |
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| 基金项目： | 国家自然科学基金资助(81570691,81270855);广东省科技计划项目(2017ZC0275);广东省科技创新战略专项资金(2019A1515012019);广东省科技发展专项资金(2017A030313715);国家卫生和计划生育委员会男性生殖与遗传重点实验室开放课题资助项目(KF201801)。 |
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| 摘 要： | 目的:探讨腺病毒载体介导神经生长因子(NGF)体外转染大鼠肌源干细胞(MDSCs)应用于基因治疗的可行性。方法:培养并纯化MDSCs细胞株，采用具有高效转染能力的腺病毒载系统将NGF基因导入MDSCs中，采用绿色荧光蛋白(EGFP)荧光技术检测转染率、实时荧光定量PCR (qPCR)检测目的基因NGF的表达情况。结果:...
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| 关 键 词： | 干细胞 遗传载体 神经生长因子 |
Investigation on transduction of nerve growth factor gene in muscle-derived stem cells mediated by adenovirus vector |
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| Fang Jianxiong,Liu Haosheng,Liu Tianqi,Zhang Zhenhui,Liu Jiumin,Pu Xiaoyong. Investigation on transduction of nerve growth factor gene in muscle-derived stem cells mediated by adenovirus vector[J]. International Journal of Urology and Nephrology, 2021, 41(1): 109-113 |
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| Authors: | Fang Jianxiong Liu Haosheng Liu Tianqi Zhang Zhenhui Liu Jiumin Pu Xiaoyong |
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| Affiliation: | (Department of Urology,Guangdong Academy of Medical Sciences,Guangdong Provincial People's Hospital,Guangzhou 510055,China) |
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| Abstract: | Objective To investigate the feasibility of gene therapy using adenovirus vector mediated nerve growth factor(NGF)gene transfection with rat muscle-derived stem cells(MDSCs)in vitro.Methods MDSCs were isolated,cultured and identified in vitro.The adenoviral vector system was utilized to introduce NGF gene into MDSCs.Expression of NGF and its downstream target proteins were detected by real-time quantitative PCR(qPCR).EGFP marker was used to determine the expression of NGF after transection.Results MDSCs were infected with adenovirus at a multiplicity of infection(MOI)of 1000 with optimal expression efficiency of 80%.EGFP marker was observed at 72 hours after transfection and then gradually enhanced.NGF gene was highly expressed in the transfected MDSCs.Conclusions The adenoviral vector carrying NGF is successfully transfected into MDSCs and stably expressed,which up-regulates the expression of target gene in MDSCs and provides a theoretical foundation for therapy of stress urinary incontinence(SUI).Moreover,this transfection method does not lead to excessive proliferation and apoptosis of MDSCs. |
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| Keywords: | Stem Cells Genetic Vectors Nerve Growth Factor |
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