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利用磁性纳米颗粒进行体外磁转染:一种用于非病毒组织工程的新型平台。

Ex vivo magnetofection with magnetic nanoparticles: a novel platform for nonviral tissue engineering.

作者信息

Yang Shieh-Yueh, Sun Jui-Sheng, Liu Cheng-Heng, Tsuang Yang-Hwei, Chen Li-Ting, Hong Chin-Yih, Yang Hong-Chang, Horng Herng-Er

机构信息

Institute of Electro-optical Science and Technology, National Taiwan Normal University, Taipei, Taiwan.

出版信息

Artif Organs. 2008 Mar;32(3):195-204. doi: 10.1111/j.1525-1594.2007.00526.x. Epub 2008 Jan 14.

Abstract

Several methods have been described to introduce DNA expression vectors into mammalian cells both in vitro and in vivo. Each system has benefits and limitations, and to date there is still no ideal method for gene transfer. In this study, we introduced a novel method of gene transfer by using Fe3O4 nanoparticles. The magnetic nanoparticles composed of Fe3O4, and the transfected genes used are Lac Z and enhanced green fluorescence protein gene (EGFG). Four different groups of preparations included in this study were homemade liposome-enveloped EGFP-DNA/Fe3O4, homemade liposome EGFP-DNA gene without magnetic Fe3O4 nanoparticles, lipofectamine 2000-enveloped EGFP-DNA, and EGFP-DNA gene only. Mice osteoblast and He99 lung cancer cell line were used as host cells for gene transfection. The time-dependent EGFP gene expression was monitored and analyzed. The results showed that the diameter of the complex was less than 100 nm. There was no cytotoxicity observed at any of the magnetic Fe3O4 nanoparticle concentrations tested. In the presence of magnetic field, the liposome-enveloped EGFP-DNA/Fe3O4 complex exhibited a much higher efficiency for transfecting EGFP-DNA into osteoblast cells under external magnetic fields. The gene can be transfected into cells with an aid of magnetic vectors and magnetic force. Under a gradient magnetic field, the efficiency of magnetofection is enhanced as compared to that without magnetic field.

摘要

已经描述了几种在体外和体内将DNA表达载体导入哺乳动物细胞的方法。每个系统都有其优点和局限性,迄今为止,仍然没有理想的基因转移方法。在本研究中,我们介绍了一种使用Fe3O4纳米颗粒进行基因转移的新方法。磁性纳米颗粒由Fe3O4组成,所使用的转染基因是Lac Z和增强型绿色荧光蛋白基因(EGFG)。本研究中包含的四组不同制剂分别是自制脂质体包裹的EGFP-DNA/Fe3O4、不含磁性Fe3O4纳米颗粒的自制脂质体EGFP-DNA基因、Lipofectamine 2000包裹的EGFP-DNA以及仅EGFP-DNA基因。小鼠成骨细胞和He99肺癌细胞系用作基因转染的宿主细胞。监测并分析了EGFP基因的时间依赖性表达。结果表明,复合物的直径小于100nm。在所测试的任何磁性Fe3O4纳米颗粒浓度下均未观察到细胞毒性。在磁场存在下,脂质体包裹的EGFP-DNA/Fe3O4复合物在外部磁场作用下将EGFP-DNA转染到成骨细胞中的效率要高得多。借助磁性载体和磁力可以将基因转染到细胞中。在梯度磁场下,与无磁场相比,磁转染效率得到提高。

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