Microfluidic electroporation for delivery of small molecules and genes into cells using a common DC power supply
Microfluidic electroporation for delivery of small molecules and genes into cells using a common DC power supply
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DOI：
10.1002/bit.21784
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发表时间：
2008-06-15
影响因子：
3.8
通讯作者：
Lu, Chang
中科院分区：
文献类型：
--
作者：
Wang, Hsiang-Yu;Lu, Chang
电穿孔是一种将药物和基因等不必要的外来分子导入细胞的有效方法。传统的电穿孔一直基于短电脉冲(电脉冲)的应用。电脉冲需要专门的设备，而且不能很容易地与基于恒定电压的电泳等技术相结合。在这里，我们演示了利用我们先前开发的基于恒定直流(DC)电压的微流控电穿孔技术将小分子和基因输送到细胞中。我们证明了两个分子进入中国仓鼠卵巢细胞(CHO-K1)：膜不透性核酸染料(SYTOX(R)Green)和携带绿色荧光蛋白基因的质粒载体(pEGFP-C1)。我们的设备可以通过不同的几何结构对流动的细胞施加场变化，类似于施加单个或多个脉冲。我们研究了器件的电气参数和不同的几何形状对转染率和细胞存活率的影响。我们的技术通过消除对脉冲发生器的需求，为基于电穿孔的药物和基因输送提供了一种简单的解决方案。我们设想，这些简单的微型电穿孔设备将有可能在微芯片平台上并行工作，这种技术将允许对药物和基因进行高通量功能筛选。
Electroporation is an efficient method of introducing foreign impermeant molecules such as drugs and genes into cells. Conventional electroporation has been based on the application of short electrical pulses (electropulsation). Electropulsation requires specialized equipment and cannot be integrated easily with techniques such as electrophoresis which is based on constant voltage. Here we demonstrate the delivery of small molecules and genes into cells, using a microfluidic electroporation technique based on constant direct current (DC) voltage that we developed earlier. We demonstrate the delivery of two molecules into Chinese hamster ovary (CHO-K1) cells: a membrane impermeable nucleic acid dye (SYTOX (R) Green) and a plasmid vector carrying the gene for green fluorescent protein (pEGFP-C1). Our devices can exert field variations to flowing cells that are analogous to the application of single or multiple pulses by having different geometries. We investigate the effects of the electrical parameters and different geometries of the device on the transfection efficiency and cell viability. Our technique provides a simple solution to electroporation-based drug and gene delivery by eliminating the need for a pulse generator. We envision that these simple microscale electroporation devices will have the potential to work in parallel on a microchip platform and such technology will allow high-throughput functional screening of drugs and genes.