摘要
该文通过体外构建三维(3D)细胞培养模型,探讨基于3D细胞模型的基因转染条件。实验选用甲基纤维素半固体培养体系,建立了体外3D细胞模型。并采用聚乙烯亚胺(PEI,25 k Da)介导荧光标记质粒和增强绿色荧光蛋白报告基因的转染,激光扫描共聚焦三维重构技术进一步分析基因在3D细胞模型中的传输及表达情况。结果表明,当PEI和质粒DNA的复合比为10时,PEI介导的基因在3D细胞模型中具有良好的传输能力和一定的表达能力。本研究结果可为3D细胞模型在基因转染方面的应用提供实验依据。
This paper aims to explore the conditions of gene transfection utilizing the three-dimensional(3D)cell culture model.Firstly,3D cell model was established by vitro using semisolid culture system of methyl cellulose.Then Cy5 labeled gene and enhanced green fluorescence protein(EGFP)report gene were transfected into 3D cell model via polyethylene imine(PEI,25 kDa).The gene delivery and EGFP expression was analyzed through 3D reconstruction method using confocal laser scanning microscope.The results showed that when the composite ratio of PEI to plasmid DNA was 10,the PEI-mediated gene had good transport ability and certain expression ability in the 3D cell model.The results of this study can provide experimental basis for the application of 3D cell model in gene transfection.
作者
刘艳红
张骥
余孝其
李静
王娜
LIU Yanhong;ZHANG Ji;YU Xiaoqi;LI Jing;WANG Na(Experiment Teaching Center of Basic Chemistry,Sichuan University,Chengdu 610064,China;College of Chemistry,Sichuan University,Chengdu 610064,China)
出处
《实验科学与技术》
2018年第6期20-23,共4页
Experiment Science and Technology
基金
四川大学实验技术项目资助(2015-44)
国家自然科学基金(21472131)
关键词
化学生物学
创新实验
3D细胞模型
基因转染
chemical biology
innovative experiment
3D cell model
gene transfection
