Electrotransfection of mammalian cells using microchannel-type electroporation chip

Shik Shin Young, Keunchang Cho, Kim Jung Kyung, Hee Lim Sun, Hee Park Chan, Baek Lee Kyu, Yongdoo Park, Chanil Chung, Dong Chul Han, Keun Chang Jun

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

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 μm 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.

Original languageEnglish
Pages (from-to)7045-7052
Number of pages8
JournalAnalytical chemistry
Volume76
Issue number23
DOIs
Publication statusPublished - 2005 Jan 3

ASJC Scopus subject areas

  • Analytical Chemistry

Fingerprint Dive into the research topics of 'Electrotransfection of mammalian cells using microchannel-type electroporation chip'. Together they form a unique fingerprint.

  • Cite this

    Young, S. S., Cho, K., Jung Kyung, K., Sun, H. L., Chan, H. P., Kyu, B. L., Park, Y., Chung, C., Han, D. C., & Jun, K. C. (2005). Electrotransfection of mammalian cells using microchannel-type electroporation chip. Analytical chemistry, 76(23), 7045-7052. https://doi.org/10.1021/ac0496291