Wang Hsiang-Yu, Lu Chang
School of Chemical Engineering, Purdue University, West Lafayette, Indiana, USA.
Biotechnol Bioeng. 2008 Jun 15;100(3):579-86. doi: 10.1002/bit.21784.
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 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.
电穿孔是一种将药物和基因等外来非渗透性分子导入细胞的有效方法。传统的电穿孔基于短电脉冲(电脉动)的应用。电脉动需要专门的设备,并且不容易与基于恒定电压的电泳等技术集成。在这里,我们展示了使用我们之前开发的基于恒定直流(DC)电压的微流体电穿孔技术,将小分子和基因导入细胞。我们展示了将两种分子导入中国仓鼠卵巢(CHO-K1)细胞:一种膜不可渗透的核酸染料(SYTOX Green)和一种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具有不同的几何形状,对流动的细胞施加类似于单个或多个脉冲应用的场变化。我们研究了电参数和设备不同几何形状对转染效率和细胞活力的影响。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递送提供了一个简单的解决方案。我们设想这些简单的微尺度电穿孔设备有可能在微芯片平台上并行工作,并且这种技术将允许对药物和基因进行高通量功能筛选。