Authors: W.G. Lee, H. Bang, H. Yun, J.A. Kim, K. Cho, Y.S. Shin, C. Chung, N.C. Chung, J.K. Chang and D.-C. Han
Affilation: Seoul National University, Korea
Pages: 243 - 246
Keywords: electroporation, electrotransfection, microstructure, mammalian cells, silica nanoparticles
Electroporation is a powerful gene transfection method to deliver foreign biological molecules such as DNA and protein into living cells with electric pulsations. However, conventional cuvette-type electroporation techniques generally suffer from poor transfection efficiency and cell survivability. We recently developed a novel method for electrotransfection using various microchannels and capillaries. The uniform electric field inside the narrow microchannels and capillaries and few side reactions owing to the small area of electrodes enabled us to enhance transfection efficiency and cell survivability. One of the advantages of microchannel-type electroporation is in-situ visualization of uptake phenomena of molecules into cells, which makes it possible to profoundly investigate the basic mechanisms of DNA transfer and intracellular response to the external electric pulses. In this study, many experimental observations at single-cell level were made to explore the uptake mechanism of small and macro molecules into mammalian cells using single- and multi-microchannel equipped on an electroporation tool kit. Several image analyses on directionality and polarity of uptake materials were performed such as propidium iodide (PI), RNA, DNA and fluorescent dye-doped silica nanoparticles (SiNPs). In conclusion, a full understanding of these uptake phenomena would be helpful to practical applications of future gene or cell therapy.
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