A novel transfection method for mammalian cells using gas plasma.
Introduction of foreign genes into target cells is a crucial step for achievement of gene therapy. We have recently developed a novel transfection system for eukaryotic cells, namely the electric pulse-activated gas plasma generator. To measure the transfection efficiency and mortality by flow-cytometry, we employed enhanced green fluorescent protein and propidium iodide staining, respectively. One day after the 1-3s plasma exposures with DNA concentration at 0.5 microg/microl, favorable transfection efficiencies (17.8-21.6%) and mortalities (0.65-2.86%) were obtained for HeLa-S3, HT-1080 and MCF-7 cells. The recipient cells became transiently permeable for plasmid DNA during the plasma exposure, suggesting that plasma-mediated transfection may involve similar mechanisms that accounts for electroporation. The relatively low mortality rates are encouraging in our attempt to apply this system to the various cell lines including the primary cell cultures.
Sakai
Yasunari
Y
Department of Pediatrics, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Fukuoka 812-8582, Japan. yasunari@pediatr.med.kyushu-u.ac.jp
Khajoee
Vahid
V
Ogawa
Yasuhiro
Y
Kusuhara
Koichi
K
Katayama
Yoshiki
Y
Hara
Toshiro
T
D045744:Cell Line, Tumor; D002470:Cell Survival; D003470:Culture Media; D004249:DNA Damage; D018274:Electroporation / Q000295:instrumentation* / Q000379:methods*; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000378:metabolism; D014162:Transfection
D003470:Culture Media