An electroporation microchip for gene transfection and system optimization

Min Li, Kuo-Chin Lin, C. Su, Yu-Tsung Wang, Chun-Sheng Fang, Yu-Cheng Lin
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引用次数: 2

Abstract

This paper describes and characterizes an electroporation microchip device and system optimization. This device combines microfabrication techniques, logic circuit, and electrophoresis design to create a multi-function gene transfection device that can be used in a number of medical science research applications. This device consists of two components: a cell-accommodation cavity and electrodes for providing electroporation and electrophoresis electric power. We demonstrated the ability to use electrophoresis force to increase the gene concentration on site-specific surface of cell lines which further enhanced the gene delivery. Meanwhile, the parameters of the electroporation system, which could have an influence on the delivery rate, were optimized by the Taguchi method, resulting in optimized values of 50 mum electrode gap, 80 mug/mL pEGFP-N1 concentration, 6 V applied transfection voltage, and 2-pulse per time with above 95 % confidence. Experimental results showed that the efficiency of gene transfection with an attracting-electric field become much higher than that without an attracting-electric field, and the delivery rate was up to 35.89 % when utilizing GFP genes into NIH-3T3 cells. The electrostatic force can be designed into specific regions, where the DNA plasmids are attracted to provide the region-targeting function. The adherent cells could be manipulated in situ without detachment by this electroporation microchip. The system has several advantages of portable, cost-effective, high transfection rate and easy operation.
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一种用于基因转染和系统优化的电穿孔芯片
本文介绍了一种电穿孔微芯片器件的特点和系统优化。该装置结合了微加工技术、逻辑电路和电泳设计,创造了一种多功能基因转染装置,可用于许多医学科学研究应用。该装置由两个部分组成:一个容纳细胞的腔和提供电穿孔和电泳电力的电极。我们证明了利用电泳力增加基因在细胞系位点特异性表面的浓度的能力,从而进一步增强了基因的传递。同时,采用田口法对影响递送率的电穿孔系统参数进行优化,得到电极间隙为50 μ m、pEGFP-N1浓度为80 μ g/mL、转染电压为6 V、2脉冲/次的最优值,置信度在95%以上。实验结果表明,有吸引电场的转染效率明显高于无吸引电场的转染效率,利用GFP基因转染NIH-3T3细胞的转染率高达35.89%。静电力可以被设计到特定的区域,在那里DNA质粒被吸引,以提供区域靶向功能。利用该微晶片可以原位操作贴壁细胞,使其不脱落。该系统具有便携、经济、转染率高、操作方便等优点。
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