TY - JOUR
T1 - A novel transfection method for mammalian cells using gas plasma
AU - Sakai, Yasunari
AU - Khajoee, Vahid
AU - Ogawa, Yasuhiro
AU - Kusuhara, Koichi
AU - Katayama, Yoshiki
AU - Hara, Toshiro
N1 - Funding Information:
We thank Drs. Teruhisa Tsuzuki, Yoshimichi Nakatsu and Kenji Ihara for critical reviews of this manuscript and helpful discussions. This work was supported by a Grant-in-Aid for Scientific Research from Ministry of Education, Culture, Sports, Science and Technology of Japan.
PY - 2006/2/10
Y1 - 2006/2/10
N2 - 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 1-3 s plasma exposures with DNA concentration at 0.5 μg/μl, 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.
AB - 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 1-3 s plasma exposures with DNA concentration at 0.5 μg/μl, 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.
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U2 - 10.1016/j.jbiotec.2005.08.020
DO - 10.1016/j.jbiotec.2005.08.020
M3 - Article
C2 - 16216376
AN - SCOPUS:30944464016
SN - 0168-1656
VL - 121
SP - 299
EP - 308
JO - Journal of Biotechnology
JF - Journal of Biotechnology
IS - 3
ER -