Electroporation microchips for in vitro gene transfection

被引:47
作者
Lin, YC [1 ]
Huang, MY [1 ]
机构
[1] Natl Cheng Kung Univ, Dept Engn Sci, Tainan 701, Taiwan
关键词
D O I
10.1088/0960-1317/11/5/315
中图分类号
TM [电工技术]; TN [电子技术、通信技术];
学科分类号
0808 ; 0809 ;
摘要
A detailed study of a novel electroporation (EP) microchip for in vitro gene transfection has been conducted. Electroporation is a technique with which DNA molecules can be delivered into cells in a chamber using high electric field pulses. Compared to a commercial electroporator, the EP microchip can attain the necessary electric field for the electroporation process but takes advantage of a much lower required voltage. The EP microchip overcomes the potential risk of using a high voltage, which is the drawback of current electroporation technology. Furthermore, the advantages of this EP microchip are the use of a very small amount of cells and genes, a very low applied voltage, a simple power supply and a much simpler cell preparation process. Any of these advantages are better than, or cannot be found in, a commercial electroporator. The EP microchip, consisting of a defined cell culture cavity region with thin-film electrodes made of titanium and gold, was fabricated on a glass slide using microfabrication technologies, which include evaporation, photolithography and wet-etching methods. Two different cell lines, Huh-7 and 293T, were used to demonstrate the transfection. The experimental results show that the EP microchip can successfully deliver green fluorescent protein genes into both cell lines. This work demonstrates that this EP microchip can provide in vitro gene transfection with a very small amount of cells and plasmids, requiring a much lower applied voltage and a simpler process than current commercial equipment. This EP microchip will have many useful applications in gene therapy.
引用
收藏
页码:542 / 547
页数:6
相关论文
共 23 条
[1]   STUDIES OF CELL PELLETS .1. ELECTRICAL-PROPERTIES AND POROSITY [J].
ABIDOR, IG ;
LI, LH ;
HUI, SW .
BIOPHYSICAL JOURNAL, 1994, 67 (01) :418-426
[2]   Assessing the potential of skin electroporation for the delivery of protein- and gene-based drugs [J].
Banga, AK ;
Prausnitz, MR .
TRENDS IN BIOTECHNOLOGY, 1998, 16 (10) :408-412
[3]  
Budker V, 2000, J GENE MED, V2, P76, DOI 10.1002/(SICI)1521-2254(200003/04)2:2<76::AID-JGM97>3.0.CO
[4]  
2-4
[5]  
Canatella P. J., 1999, Proceedings of the First Joint BMES/EMBS Conference. 1999 IEEE Engineering in Medicine and Biology 21st Annual Conference and the 1999 Annual Fall Meeting of the Biomedical Engineering Society (Cat. No.99CH37015), DOI 10.1109/IEMBS.1999.802124
[6]   Immunization of rabbits with cottontail rabbit papillomavirus E1 and E2 genes: protective immunity induced by gene gun-mediated intracutaneous delivery but not by intramuscular injection [J].
Han, R ;
Reed, CA ;
Cladel, NM ;
Christensen, ND .
VACCINE, 2000, 18 (26) :2937-2944
[7]   ELECTRONIC GENETIC - PHYSICAL AND BIOLOGICAL ASPECTS OF CELLULAR ELECTROMANIPULATION [J].
HOFMANN, GA ;
EVANS, GA .
IEEE ENGINEERING IN MEDICINE AND BIOLOGY MAGAZINE, 1986, 5 (04) :6-25
[8]   Micro-Electroporation: Improving the Efficiency and Understanding of Electrical Permeabilization of Cells [J].
Huang, Yong ;
Rubinsky, Boris .
BIOMEDICAL MICRODEVICES, 1999, 2 (02) :145-150
[9]   TRANSFECTION BY ELECTROPORATION - CELL GENE-TRANSFER USING ELECTRICAL IMPULSES, A NEW PROCESS IN BLOOD CANCER-RESEARCH [J].
JAMIESON, RD ;
BODGER, MP ;
BODGER, PS ;
BARAN, D .
IEE PROCEEDINGS-A-SCIENCE MEASUREMENT AND TECHNOLOGY, 1989, 136 (01) :41-44
[10]  
JAYARAM S, 1991, IND APPL SOC ANN M P, V1, P674