Study of gene transfection enhancement and Parameters Optimization using microfluidic electroporation chip

We developed a simple fabrication, low voltage, low power consumption and easy way of enhancing gene transfection efficiency: combining high and low electric fields, adjusting the voltage, pulse number, duration, buffer and conductivity. The 293T cells (Human Embryonic Kidney cells) were tested. The optimum combining condition was a high electric field 1600 V/cm for 0.6 ms x 2 and low electric field 800 V/cm for 1.2 ms x 3. It resulted in a survival rate larger than 70%, and the transfection rate was about 45%. The buffer of the largest transfection rate was Hypoosmolar. The conductivity of the buffer was an important parameter, and the appropriate value was 1.0-3.6 mS/cm. The transfection rate of 293T cells using Cytoporation buffer could be enhanced by adding potassium ion, and the volume ratio should be lower than 2%.