Flow-through Electroporation for Transfection Based On Low-Frequency AC Voltage

Electroporation has been demonstrated to be an efficient technique for gene transfection of mammalian cells. Flow-through electroporation facilitates high throughput processing of large numbers of cells. However, the employment of direct current (DC) voltage produces electrolysis of water and bubbles with high field intensities and these problems lower the cell viability and prevent using of very high field intensities. We demonstrate here flow-through electroporation based on low-frequency alternating current (AC) voltage (27-4870Hz). We discovered that the transfection based on 379Hz AC voltage generated higher efficiency than that using DC voltage. We systematically studied the effects of the AC frequency on the cell size expansion, permeability, transfection efficiency and cell viability. Our study provides a alternative means for highly efficiency gene delivery and generates new insights into AC based electroporation.