A novel transfection method for mammalian cells using gas plasma
A novel transfection method for mammalian cells using gas plasma
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DOI：
10.1016/j.jbiotec.2005.08.020
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发表时间：
2006-02-10
影响因子：
4.1
通讯作者：
Hara, T
中科院分区：
文献类型：
--
作者：
Sakai, Y;Khajoee, V;Hara, T
将外源基因导入靶细胞是实现基因治疗的关键步骤。我们最近开发了一种新的真核细胞转染系统，即电脉冲激活气体等离子体发生器。为了通过流式细胞术测量转染效率和死亡率，我们分别采用增强型绿色荧光蛋白和碘化丙啶染色。在0.5 μ g/μ l的DNA浓度下，1-3秒的血浆暴露后1天，对于HeLa-S3、HT-1080和MCF-7细胞获得了有利的转染效率（17.8-21.6%）和死亡率（0.65-2.86%）。受体细胞在等离子体暴露期间变得瞬时可渗透质粒DNA，表明等离子体介导的转染可能涉及类似的机制，解释电穿孔。相对较低的死亡率是令人鼓舞的，我们试图将该系统应用于各种细胞系，包括原代细胞培养。(c)2005 Elsevier B. V.保留所有权利。
introduction of foreign genes into target cells is a crucial step for achievement of gene therapy. We have recently developed a novel transfection system for eukaryotic cells, namely the electric pulse-activated gas plasma generator. To measure the transfection efficiency and mortality by flow-cytometry, we employed enhanced green fluorescent protein and propidium iodide staining, respectively. One day after the 1-3 s plasma exposures with DNA concentration at 0.5 mu g/mu l, favorable transfection efficiencies (17.8-21.6%) and mortalities (0.65-2.86%) were obtained for HeLa-S3, HT-1080 and MCF-7 cells. The recipient cells became transiently permeable for plasmid DNA during the plasma exposure, suggesting that plasma-mediated transfection may involve similar mechanisms that accounts for electroporation. The relatively low mortality rates are encouraging in our attempt to apply this system to the various cell lines including the primary cell cultures. (c) 2005 Elsevier B.V. All rights reserved.