Determination of optimum gene transfection conditions using the Taguchi method for an electroporation microchip

This study demonstrates the optimization of an electroporation (EP) microchip system for gene transfection using the Taguchi method. The computer-aided engineering simulations show that approximately 80% of the cells are distributed in the middle region of the reaction chamber and are subjected to the applied electric field. A smaller electrode gap, such as 50 mu m, gained a larger and more uniform electric field distribution in the middle region. The parameters of the EP microchip system, which can have an influence on the transfecting rate were optimized, resulting in values of 50 mu m, electrode gap, 20 mu g/mL pEGFP-N1 concentration, 6 V applied voltage, and 2-pulse. A larger-the-better Omega transformation was employed for better additivity. The analyzed results of interactions A x C, A x D, and C x D show that there were strong interactions on level 3 of factors A, C, and D among each other. Using the optimized EP microchip system, the great majority of pEGFP-N1 plasmids was successfully transfected into the NIH Swiss mouse embryo fibroblast cell lines (NIH-3T3). The optimized EP parameters helped with improving the EP microchip system and elevating the average transfection rate from 17.2 to 40.36%, and the inaccuracy of the predicted transfection rate of 43.67% was as small as 3.31%. (C) 2007 Elsevier B.V. All rights reserved.