Geng Tao, Zhan Yihong, Lu Chang
Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN 47906, USA. tgeng@ purdue.edu
Annu Int Conf IEEE Eng Med Biol Soc. 2012;2012:2579-82. doi: 10.1109/EMBC.2012.6346491.
Electroporation is one of the most widely used physical methods to deliver exogenous nucleic acids into cells with high efficiency and low toxicity. Conventional electroporation systems typically require expensive pulse generators to provide short electrical pulses at high voltage. In this work, we demonstrate a flow-through electroporation method for continuous transfection of cells based on disposable chips, a syringe pump, and a low-cost power supply that provides a constant voltage. We successfully transfect cells using either DC or AC voltage with high flow rates (ranging from 40 µl/min to 20 ml/min) and high efficiency (up to 75%). We also enable the entire cell membrane to be uniformly permeabilized and dramatically improve gene delivery by inducing complex migrations of cells during the flow.
电穿孔是将外源核酸高效、低毒地导入细胞的最广泛使用的物理方法之一。传统的电穿孔系统通常需要昂贵的脉冲发生器来提供高电压的短电脉冲。在这项工作中，我们展示了一种基于一次性芯片、注射泵和提供恒定电压的低成本电源的用于细胞连续转染的流通式电穿孔方法。我们使用直流或交流电压在高流速（范围从40微升/分钟到20毫升/分钟）下成功转染细胞，且效率高达75%。我们还能使整个细胞膜均匀通透，并通过在流动过程中诱导细胞的复杂迁移显著改善基因递送。
