Document Detail


Electrotransfection of mammalian cells using microchannel-type electroporation chip.
MedLine Citation:
PMID:  15571358     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Young Shik Shin; Keunchang Cho; Jung Kyung Kim; Sun Hee Lim; Chan Hee Park; Kyu Baek Lee; Yongdoo Park; Chanil Chung; Dong-Chul Han; Jun Keun Chang
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Analytical chemistry     Volume:  76     ISSN:  0003-2700     ISO Abbreviation:  Anal. Chem.     Publication Date:  2004 Dec 
Date Detail:
Created Date:  2004-12-01     Completed Date:  2006-07-26     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  7045-52     Citation Subset:  IM    
Affiliation:
School of Mechanical and Aerospace Engineering, Seoul National University, 312-203, San 56-1, Shinlim-dong, Kwanak-gu, Seoul, 151-742, Korea.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Line, Tumor
Cell Membrane Permeability / physiology
Dimethylpolysiloxanes
Electroporation / instrumentation,  methods*
Equipment Design
Gene Expression Regulation
Green Fluorescent Proteins / chemistry,  genetics
Humans
Microfluidic Analytical Techniques / instrumentation,  methods*
Scyphozoa / chemistry
Sensitivity and Specificity
Silicones
Surface Properties
Time Factors
Transfection / methods*
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Dimethylpolysiloxanes; 0/Silicones; 0/enhanced green fluorescent protein; 147336-22-9/Green Fluorescent Proteins; 63148-62-9/baysilon

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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