title: Novel Parallelized Electroporation by Electrostatic Manipulation of a Water-in-Oil Droplet as a Microreactor.
abstract: Electroporation is the most widely used transfection method for delivery of cell-impermeable molecules into cells. We developed a novel gene transfection method, water-in-oil (W/O) droplet electroporation, using dielectric oil and an aqueous droplet containing mammalian cells and transgene DNA. When a liquid droplet suspended between a pair of electrodes in dielectric oil is exposed to a DC electric field, the droplet moves between the pair of electrodes periodically and droplet deformation occurs under the intense DC electric field. During electrostatic manipulation of the droplet, the local intense electric field and instantaneous short circuit via the droplet due to droplet deformation facilitate gene transfection. This method has several advantages over conventional transfection techniques, including co-transfection of multiple transgene DNAs into even as few as 103 cells, transfection into differentiated neural cells, and the capable establishment of stable cell lines. In addition, there have been improvements in W/O droplet electroporation electrodes for disposable 96-well plates making them suitable for concurrent performance without thermal loading by a DC electric field. This technique will lead to the development of cell transfection methods for novel regenerative medicine and gene therapy.
authors.affiliation: Department of Environmental and Life Sciences, Toyohashi University of Technology, Toyohashi, Aichi, Japan.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Kurita
authors.affiliation: Department of Environmental and Life Sciences, Toyohashi University of Technology, Toyohashi, Aichi, Japan.
authors.forename: Shota
authors.initials: S
authors.lastname: Takahashi
authors.affiliation: Department of Environmental and Life Sciences, Toyohashi University of Technology, Toyohashi, Aichi, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Asada
authors.affiliation: Department of Environmental and Life Sciences, Toyohashi University of Technology, Toyohashi, Aichi, Japan.
authors.forename: Minako
authors.initials: M
authors.lastname: Matsuo
authors.affiliation: Department of Environmental and Life Sciences, Toyohashi University of Technology, Toyohashi, Aichi, Japan.
authors.forename: Kenta
authors.initials: K
authors.lastname: Kishikawa
authors.affiliation: Department of Environmental and Life Sciences, Toyohashi University of Technology, Toyohashi, Aichi, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Mizuno
authors.affiliation: Department of Environmental and Life Sciences, Toyohashi University of Technology, Toyohashi, Aichi, Japan.|Electronics-Inspired Interdisciplinary Research Institute (EIIRIS), Toyohashi University of Technology, Toyohashi, Aichi, Japan.
authors.forename: Rika
authors.initials: R
authors.lastname: Numano
mesh_terms: D000818:Animals; D002460:Cell Line; D004247:DNA / Q000235:genetics*; D018274:Electroporation; D008322:Mammals; D055672:Static Electricity; D014162:Transfection / Q000379:methods*; D019076:Transgenes; D014867:Water
chemical_list: D014867:Water; D004247:DNA