Permeabilization of Adhered Cells Using an Inert Gas Jet

Abstract

Various cell transfection techniques exist and these can be broken down to three broad categories: viral, chemical and mechanical. This protocol describes a mechanical method to temporally permeabilize adherent cells using an inert gas jet that can facilitate the transfer of normally non-permeable macromolecules into cells. We believe this technique works by imparting shear forces on the plasma membrane of adherent cells, resulting in the temporary formation of micropores. Once these pores are created, the cells are then permeable to genetic material and other biomolecules. The mechanical forces involved do run the risk of permanently damaging or detaching cells from their substrate. There is, therefore, a narrow range of inert gas dynamics where the technique is effective. An inert gas jet has proven efficient at permeabilizing various adherent cell lines including HeLa, HEK293 and human abdominal aortic endothelial cells. This protocol is appropriate for the permeabilization of adherent cells both in vitro and, as we have demonstrated, in vivo, showing it may be used for research and potentially in future clinical applications. It also has the advantage of permeabilizing cells in a spatially restrictive manner, which could prove to be a valuable research tool.

The assessment of cold atmospheric plasma treatment of DNA in synthetic models of tissue fluid, tissue and cells

“…This would explain only minimal leakage of the encapsulated CF from the vesicles with plasma jet treatment. (3) The gas flow itself imparts greater mechanical forces onto the phospholipid membrane of the vesicles [100]. Consequently, greater mechanical forces exerted onto the phospholipid membrane by the higher flow rate plasma jet, enhanced the ingress of RONS.…”

Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts

“…Intense pulses of fluid shear can be directed at cells by firing jets of pressurized inert gas toward them 787,788 . Similar to the case of cone-plate viscometers, it was found that excessive shear forces can rip cells from the underlying substrate, but if modulated just below this range, were capable of permeabilizing cell membranes while leaving adherent cells in place.…”

“…With the appropriate optimizations, intracellular delivery of dextrans, plasmids, and other cargo has been demonstrated in common adherent cell lines permeabilized by inert gas jets. 825,826 6.1.12. Fluid Shear: Sonoporation.…”

“…Intense pulses of fluid shear can be directed at cells by firing jets of pressurized inert gas toward them. , Similar to the case of cone–plate viscometers, it was found that excessive shear forces can rip cells away from the underlying substrate, but if modulated just below this range, they were capable of permeabilizing cell membranes while leaving adherent cells in place. With the appropriate optimizations, intracellular delivery of dextrans, plasmids, and other cargo has been demonstrated in common adherent cell lines permeabilized by inert gas jets. , …”