Electrotransfection of mammalian cells using microchannel-type electroporation chip.
Transfection of DNA molecules into mammalian cells with electric pulsations, which is so-called electroporation, is a powerful and widely used method that can be directly applied to gene therapy. However, very little is known about the basic mechanisms of DNA transfer and cell response to the electric pulse. We developed a microelectroporation chip with poly(dimethylsiloxane) (PDMS) to investigate the mechanism of electroporation as a first step of DNA transfer and to introduce the benefits of miniaturization into the genetic manipulation. The microelectroporation chip has a microchannel with a height of 20 microm and a length of 2 cm. Owing to the transparency of PDMS, we could in situ observe the uptake process of propidium iodide (PI) into SK-OV-3 cells, which shows promise in visualization of gene delivery in living cells. We also noticed the geometric effect on the degree of electroporation in microchannels with diverse channel width. This experimental result shows that the geometry can be another parameter to be considered for the electroporation when it is performed in microchannels with an exponential decaying pulse generator. Cell culturing is possible within the microelectroporation chip, and we also successfully transfected SK-OV-3 cells with enhanced green fluorescent protein genes, which demonstrates the feasibility of the microelectroporation chip in genetic manipulation.
Shin
Young Shik
YS
School of Mechanical and Aerospace Engineering, Seoul National University, 312-203, San 56-1, Shinlim-dong, Kwanak-gu, Seoul, 151-742, Korea.
Cho
Keunchang
K
Kim
Jung Kyung
JK
Lim
Sun Hee
SH
Park
Chan Hee
CH
Lee
Kyu Baek
KB
Park
Yongdoo
Y
Chung
Chanil
C
Han
Dong-Chul
DC
Chang
Jun Keun
JK
D000818:Animals; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000502:physiology; D004129:Dimethylpolysiloxanes; D018274:Electroporation / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D037401:Scyphozoa / Q000737:chemistry; D012680:Sensitivity and Specificity; D012828:Silicones; D013499:Surface Properties; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured
D004129:Dimethylpolysiloxanes; D012828:Silicones; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C013830:baysilon