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A novel transfection method for mammalian cells using gas plasma
Yasunari Sakai1, Vahid Khajoee, Yasuhiro Ogawa
1Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Fukuoka 812-8582, Japan. yasunari@pediatr.med.kyushu-u.ac.jp
Journal of Biotechnology
|October 12, 2005
Summary
A new electric pulse-activated gas plasma generator offers an efficient method for gene therapy. This novel system achieves high transfection efficiencies with low cell mortality in various eukaryotic cell lines.
Area of Science:
- Biotechnology
- Cell Biology
- Gene Therapy
Background:
- Gene therapy relies on efficient foreign gene delivery into target cells.
- Existing transfection methods can have limitations in efficiency and cell viability.
Purpose of the Study:
- To introduce a novel electric pulse-activated gas plasma generator for eukaryotic cell transfection.
- To evaluate the transfection efficiency and cell mortality of this new system.
Main Methods:
- Utilized an electric pulse-activated gas plasma generator for gene delivery.
- Employed enhanced green fluorescent protein (eGFP) for transfection efficiency measurement.
- Used propidium iodide staining and flow cytometry to assess cell mortality.
- Tested on HeLa-S3, HT-1080, and MCF-7 cell lines with DNA concentration at 0.5 microg/microl.
Main Results:
- Achieved favorable transfection efficiencies ranging from 17.8% to 21.6%.
- Observed low cell mortalities between 0.65% and 2.86% after 1-3s plasma exposures.
- Demonstrated transient cell permeability to plasmid DNA during plasma exposure.
- Successfully transfected multiple eukaryotic cell lines.
Conclusions:
- The electric pulse-activated gas plasma generator is a promising tool for gene delivery.
- The system shows potential for application in various cell lines, including primary cultures, due to low mortality.
- Plasma-mediated transfection may share mechanisms with electroporation.