An epoch-making application of discharge plasma phenomenon to gene-transfer
An epoch-making application of discharge plasma phenomenon to gene-transfer
复制标题
DOI：
10.1002/bit.20659
复制
发表时间：
2005-12-30
影响因子：
3.8
通讯作者：
Nishimura, S
中科院分区：
文献类型：
--
作者：
Ogawa, Y;Morikawa, N;Nishimura, S
我们发现了利用放电等离子体的划时代基因转移方法。尽管目前的基因转移实验普遍使用电穿孔方法，但其不能将基因充分转移至原代细胞中。本研究中采用的大气压放电等离子体最初用于非生物材料的表面处理。我们假设它可以在靶细胞表面提供适当的作用，并将其应用于将基因转移到各种类型的细胞中。等离子体技术成功地将绿色荧光蛋白（GFP）质粒有效转移到从大鼠大脑皮质获得的有丝分裂后神经元细胞中，而传统设备的电穿孔无法充分地将基因转移到其中，因为细胞是附着的。通过血浆处理将GFP基因转染大鼠嗜铬细胞瘤PC12细胞后，细胞保留了其功能，即神经生长因子诱导的分化。此外，等离子体技术的基因转移也适用于其他类型的细胞系，例如作为贴壁细胞系的HeLa细胞和中国仓鼠肺（CHL）细胞，作为悬浮细胞系的Jurkat细胞，以及另一种类型的原代细胞，人脐静脉内皮细胞（HUVEC）。总之，等离子体法是一种划时代的基因转移技术，可以有效地将基因转移到电穿孔无法转移基因的原代细胞中。此外，该方法能够将基因普遍转移到各种类型的细胞中，同时保持细胞的功能。 (c) 2005 年 Wiley 期刊公司。
We discovered an epoch-making gene transfer method utilizing discharge plasma. Although an electroporation method is commonly used in present gene transfer experiments, it cannot transfer genes into primary cells sufficiently. The atmospheric pressure discharge plasma employed in this study was originally used for surface treatment of non-biological materials. We hypothesized that it could provide a suitable effect on the surface of target cells and applied it to gene transfer into various types of cells. The plasma technology succeeded in the efficient transfer of green fluorescence protein (GFP) plasmid into post-mitotic neuronal cells obtained from cerebral cortices of rats, into which an electroporation with conventional equipment cannot transfer genes sufficiently, as the cells were attached. After the transfection of rat pheochromocytoma PC12 cells with the GFP gene by plasma treatment, the cells retained their function, that is, nerve growth factor-induced differentiation. Furthermore, gene transfer with the plasma technology was also applicable to other types of cell lines such as HeLa cells and Chinese hamster lung (CHL) cells as adherent cell lines, and Jurkat cells as a suspended cell line, and another type of primary cell, human umbilical vein endothelial cells (HUVEC). In conclusion, the plasma method is an epoch-making gene transfer technology which efficiently transfers genes into primary cells into which electroporation cannot transfer genes. Moreover, the method is able to universally transfer genes into various types of cells as the function of the cells was maintained. (c) 2005 Wiley Periodicals, Inc.