An epoch‐making application of discharge plasma phenomenon to gene‐transfer

Abstract

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.

The Necessity of Radicals for Gene Transfection by Discharge Plasma Irradiation

“…Such a technique would be expected to play an important role in advanced medical technologies like gene therapy and regenerative medicine through establishment of iPS (induced pluripotent stem) cells. A technique that uses discharge plasma irradiation was invented by Satoh, who is one of the authors, and his group in 2002 [4] and was reported by Ogawa in 2005 [5]. Since this technique is free from adverse effect associated with viruses, there are no risks as the others mentioned above.…”

Intracellular ROS mediates gas plasma-facilitated cellular transfection in 2D and 3D cultures

“…Examples of its translation into major healthcare innovations include tissue ablation and coagulation, disinfection of food and living tissues, wound healing, and even cancer treatment23242526. Several groups have reported that plasma jet with pulsed high frequency could increase the cell permeability and delivery of GFP vectors in mammalian cells such as HeLa cells, CHL cells, Jurkat cells, Hacat cells and MCF-7 cells272829. Leduc et al .…”

Electrogenetherapy of B16.F10 murine melanoma tumors with an interleukin-28 expressing DNA plasmid

“…One investigation described the use of plasma to deliver DNA into primary neuronal cells in vitro. 24 Another study by Connolly et al demonstrated increased transfection of a luciferase-expressing plasmid in the skin when the injection area was concomitantly exposed to helium plasma. 21 It has been recently demonstrated in several studies that helium plasma exposure enhances the vaccination efficacy of a plasmid-expressing HIV-1 gp120 (envelope glycoprotein) in mice when compared with injection only.…”