| 原代大鼠肾小球系膜细胞最佳电转染条件的探索和验证 |
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| 引用本文： | 徐锋,吕杨,谢院生,李晓帆,崔文鹏,苗里宁,陈香美. 原代大鼠肾小球系膜细胞最佳电转染条件的探索和验证[J]. 压电与声光, 2010, 0(5) |
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| 作者姓名： | 徐锋 吕杨 谢院生 李晓帆 崔文鹏 苗里宁 陈香美 |
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| 作者单位： | 解放军总医院全军肾脏病研究所肾脏病重点实验室;吉林大学第二医院肾病内科; |
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| 基金项目： | 国家自然科学基金项目(30630033,20971377)~~ |
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| 摘 要： | 目的探索并验证原代大鼠肾小球系膜细胞最佳电转染条件。方法设立不同的电压与电容组合、电击前孵育温度、质粒DNA浓度等电转染条件,将绿色荧光蛋白表达质粒pEGFP-C1导入原代大鼠肾小球系膜细胞。24h后比较各组细胞存活率和绿色荧光蛋白表达情况,确立最佳电转染条件。再以此条件电转染质粒pcDNA3.1和pcDNA3.1-FHL2,48h后收取两组细胞RNA和蛋白。应用RT-PCR和Western印迹检测目的基因FHL2的核酸和蛋白表达,验证电转染效果。结果在电击前室温孵育,40μg质粒,340V电压、550μF电容的最佳电转染条件下,原代大鼠肾小球系膜细胞的电转染效率可达40%以上,细胞存活率在50%左右。转染FHL2质粒组较对照组检测FHL2的核酸和蛋白表达水平明显升高(P<0.05)。结论适当的电转染条件可以有效转染原代大鼠肾小球系膜细胞。
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| 关 键 词： | 转染 电穿孔 肾小球系膜细胞 |
Optimal electric transfection conditions in primary glomerular mesangial cells |
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| XU Feng,LV Yang,XIE Yuan-sheng,LI Xiao-fan,CUI Wen-peng,MIAO Li-ning,CHEN Xiang-mei Institute of Nephrology,Chinese PLA kidney center key Laboratory,Chinese PLA General Hospital,Beijing,China. Optimal electric transfection conditions in primary glomerular mesangial cells[J]. Piezoelectrics & Acoustooptics, 2010, 0(5) |
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| Authors: | XU Feng LV Yang XIE Yuan-sheng LI Xiao-fan CUI Wen-peng MIAO Li-ning CHEN Xiang-mei Institute of Nephrology Chinese PLA kidney center key Laboratory Chinese PLA General Hospital Beijing China |
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| Affiliation: | XU Feng1,2,LV Yang1,XIE Yuan-sheng1,LI Xiao-fan1,CUI Wen-peng1,MIAO Li-ning2,CHEN Xiang-mei1 1Institute of Nephrology,Chinese PLA kidney center key Laboratory,Chinese PLA General Hospital,Beijing 100853,China,2Department of Nephrology,Second Hospital of Jilin University,Changchun 130041,Jilin Province |
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| Abstract: | Objective To study and validate the effective electric transfection conditions in primary glomerular mesangial cells of rats.Methods Twenty-four hours after green fluorescent protein(GFP) expression plasmid pEGFP-C1 was transfected into primary glomerular mesangial cells of rats under different electric pressures and capacities,incubation temperature,and plasmid DNA concentration,survival of cells and expression of green fluorescent protein were compared to determine the optical electric transfection condit... |
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| Keywords: | Transfection Electroporation Mesangial Cells |
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