An epoch-making application of discharge plasma phenomenon to gene-transfer.
We discovered an epoch-making gene transfer method utilizing discharge plasma. Although an electroporation method is commonly used in present gene transfer experiments, it cannot transfer genes into primary cells sufficiently. The atmospheric pressure discharge plasma employed in this study was originally used for surface treatment of non-biological materials. We hypothesized that it could provide a suitable effect on the surface of target cells and applied it to gene transfer into various types of cells. The plasma technology succeeded in the efficient transfer of green fluorescence protein (GFP) plasmid into post-mitotic neuronal cells obtained from cerebral cortices of rats, into which an electroporation with conventional equipment cannot transfer genes sufficiently, as the cells were attached. After the transfection of rat pheochromocytoma PC12 cells with the GFP gene by plasma treatment, the cells retained their function, that is, nerve growth factor-induced differentiation. Furthermore, gene transfer with the plasma technology was also applicable to other types of cell lines such as HeLa cells and Chinese hamster lung (CHL) cells as adherent cell lines, and Jurkat cells as a suspended cell line, and another type of primary cell, human umbilical vein endothelial cells (HUVEC). In conclusion, the plasma method is an epoch-making gene transfer technology which efficiently transfers genes into primary cells into which electroporation cannot transfer genes. Moreover, the method is able to universally transfer genes into various types of cells as the function of the cells was maintained.
Ogawa
Yasuhiro
Y
Astellas Pharma, Inc., 10-2, Kanda Tomiyama-cho, Chiyoda-ku, Tokyo 101-0043, Japan. yasuhiro.ogawa@jp.astellas.com
Morikawa
Noriyuki
N
Ohkubo-Suzuki
Akiko
A
Miyoshi
Sousuke
S
Arakawa
Hiroyuki
H
Kita
Yasuhiro
Y
Nishimura
Shintaro
S
D000818:Animals; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D016716:PC12 Cells; D010957:Plasmids; D051381:Rats