Analysis of cell exposed to non-thermal atmospheric pressure plasma for effective gene transfer

Non-thermal atmospheric pressure plasma (NTAPP) has been used in biomedical researches [1]. One of its attractive applications is gene transfection for affecting cell fate. To reveal the mechanism of gene transfer using NTAPP, surface morphology of cells exposed to NTAPP was analyzed.



Murine fibroblast cells (L929) were exposed to NTAPP using a plasma jet source (He 2 slm; 9 kV; 12. 5 kHz) and fluorescent reagent (DiYO-1) was supplied to the cells. After 1 h incubation, the cells were observed by fluorescent microscopy.



NTAPP exposure more than 13 s introduced DiYO-1 into the cells. Then, glutaraldehyde was supplied to the cells for fixing their surface morphology. The cells were freeze-dried for scanning electron microscopy. For the cells exposed to NTAPP, pores were found on the cell membrane.



To exclude the possibility that the pores were formed during freeze-drying, the cells were observed in liquid medium using scanning ion microscopy (SICM) [2]. SICM revealed pores on the cell membrane. The pores formed by NTAPP exposure can play important roles in gene delivery.



[1] S. Kumagai et al., Jpn. J. Apple. Phys., 55, 01AF01 (2016) (OPEN ACCESS); https://doi.org/10.7567/JJAP.55.01AF01

[2] S. Watanabe et al., Rev. Sci. Instrum. 90, 123704 (2019); https://doi.org/10.1063/1.5118360