A detailed study of a novel electroporation (EP) microchip for in vitro gene transfection has been conducted. Electroporation is a technique with which DNA molecules can be delivered into cells in a chamber using high electric field pulses. Compared to a commercial electroporator, the EP microchip can attain the necessary electric field for the electroporation process but takes advantage of a much lower required voltage. The EP microchip overcomes the potential risk of using a high voltage, which is the drawback of current electroporation technology. Furthermore, the advantages of this EP microchip are the use of a very small amount of cells and genes, a very low applied voltage, a simple power supply and a much simpler cell preparation process. Any of these advantages are better than, or cannot be found in, a commercial electroporator. The EP microchip, consisting of a defined cell culture cavity region with thin-film electrodes made of titanium and gold, was fabricated on a glass slide using microfabrication technologies, which include evaporation, photolithography and wet-etching methods. Two different cell lines, Huh-7 and 293T, were used to demonstrate the transfection. The experimental results show that the EP microchip can successfully deliver green fluorescent protein genes into both cell lines. This work demonstrates that this EP microchip can provide in vitro gene transfection with a very small amount of cells and plasmids, requiring a much lower applied voltage and a simpler process than current commercial equipment. This EP microchip will have many useful applications in gene therapy.
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