PEI-质粒纳米粒子对体外培养新生C57BL/6J小鼠耳蜗基底膜转染效果观察

Polyethylenimine for Plasmid Delivery to the Basilar Membrane of the Neonatal C57BL/6J Mice Cochlea in Vitro

刘永泽;周函;钱晓云;朱光洁;罗熠;李奇峰;陈杰;马登滨;高下;

1:南京大学医学院附属鼓楼医院耳鼻咽喉头颈外科

2:南京大学生命科学院

摘要
目的观察以25kDa线性聚乙稀亚胺(linear plyethylenimine,L-PEI)和分枝PEI(branched PEI,B-PEI)作为表达绿色荧光蛋白的质粒pEGFP-C1的载体,转染体外培养新生C57BL/6J小鼠基底膜的效果。方法25kDa线性或分枝PEI与质粒在5%葡萄糖溶液或PBS溶液中形成纳米粒子后,行透射电镜观察纳米粒子表征;按相同方法制备纳米粒子,行凝胶阻滞实验并在培养的293T细胞中行MTT毒性测试,选择毒性较小的PEI类型作为质粒载体在293T细胞检测其转染效果;以最适条件用其转染体外培养新生C57BL/6J小鼠耳蜗基底膜并行荧光镜观察。结果在相同条件下,B-PEI转染能力略强于L-PEI,但L-PEI毒性明显低于B-PEI;在PBS中制备的L-PEI-pDNA纳米粒子的毒性小于在葡萄糖溶液中制备的该粒子毒性;L-PEI-pDNA纳米粒子在适宜浓度范围可以转染体外培养新生小鼠耳蜗基底膜的螺旋缘纤维细胞、螺旋神经元、支持细胞,但转染效率不高。结论 L-PEI可作为转染新生小鼠耳蜗基底膜细胞的非病毒载体,但需要进一步优化获得更好的转染效果。
关键词
非病毒载体;线性/分枝聚乙稀亚胺;基因转染;耳蜗;小鼠
基金项目(Foundation):
江苏省自然科学基金(BK2010104)资助
作者
刘永泽;周函;钱晓云;朱光洁;罗熠;李奇峰;陈杰;马登滨;高下;
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