摘要
背景:新型阳离子纳米材料叶酸聚乙二醇接枝支化聚乙烯亚胺易于合成和改性,稳定性好,结构及性能容易调控,已被作为基因治疗转染载体应用于多种疾病的研究。目的:观察叶酸聚乙二醇接枝支化聚乙烯亚胺转染体外培养小鼠内耳螺旋神经节细胞的可行性及特点。方法:分别以叶酸聚乙二醇接枝支化聚乙烯亚胺(实验组)、阳离子脂质体Lipofectamine?2000(对照组)作为载体,制备携带增强型绿色荧光蛋白的转染载体系统。将昆明小鼠螺旋神经节细胞分别培养于含实验组和对照组转染载体系统的DMEM/F-12培养基中,培养24 h后,MTT法观察载体系统的细胞毒性,倒置荧光显微镜观察细胞形态及绿色荧光蛋白表达强度,流式细胞仪检测细胞转染率。结果与结论:两种载体分别转染后,螺旋神经节细胞形态均未发生明显改变,但两组载体对螺旋神经节细胞均有一定的毒性,实验组载体毒性较小且转染效率较高(P<0.05)。结果说明叶酸聚乙二醇接枝支化聚乙烯亚胺基因转染体外螺旋神经节细胞的转染效率及细胞毒性均优于传统的阳离子脂质体。
BACKGROUND：A new kind of polyethyenimine-polyethylene glycol （PEI-PEG） is apt to synthesis and modification, and has good stability, easily regulated structure and properties, which has been applied to a variety of diseases as gene transfer vectors.
OBJECTIVE：To study the feasibility and the characteristics of nanoparticles PEI-PEG as gene transfer vector for spiral ganglion cells of the inner ear of mice.
METHODS：Using PEI-PEG （experimental group） and lipofectamine^TM 2000 （control group） as gene transfer vectors, with enhanced green fluorescent protein （EGFP） as tracing protein, spiral ganglion cells were transfected in vitro, the transfection rate and mean fluorescence strength were detected by fluorescence microscopy and flow cytometry. Toxicity effects of each vector on spiral ganglion cells were determined by spectroscopic measurement of MTT method.
RESULTS AND CONCLUSION：After transfection with two vectors, spiral ganglion cells had no changes in morphology. The transfection rate of PEI-PEG was statistical y higher than that of liposome （P〈0.05）. Also, the toxicity effects of PEI-PEG to spiral ganglion cells was lightly than that of liposome （P〈0.05）. As a new gene transfer vector, PEI-PEG has a higher transfection rate and lower toxicity effects to spiral ganglion cells compared to liposome, and can serve as gene transferring system into cochlear nervous system.
出处
《中国组织工程研究》
CAS
CSCD
2014年第21期3345-3349,共5页
Chinese Journal of Tissue Engineering Research
基金
广东省自然科学基金(博士启动)项目(S2011040003553)~~
