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Novel Parallelized Electroporation by Electrostatic Manipulation of a Water-in-Oil Droplet as a Microreactor

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...

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Autores principales: Kurita, Hirofumi, Takahashi, Shota, Asada, Atsushi, Matsuo, Minako, Kishikawa, Kenta, Mizuno, Akira, Numano, Rika
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4674099/
https://www.ncbi.nlm.nih.gov/pubmed/26649904
http://dx.doi.org/10.1371/journal.pone.0144254
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author Kurita, Hirofumi
Takahashi, Shota
Asada, Atsushi
Matsuo, Minako
Kishikawa, Kenta
Mizuno, Akira
Numano, Rika
author_facet Kurita, Hirofumi
Takahashi, Shota
Asada, Atsushi
Matsuo, Minako
Kishikawa, Kenta
Mizuno, Akira
Numano, Rika
author_sort Kurita, Hirofumi
collection PubMed
description 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 10(3) 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.
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spelling pubmed-46740992015-12-23 Novel Parallelized Electroporation by Electrostatic Manipulation of a Water-in-Oil Droplet as a Microreactor Kurita, Hirofumi Takahashi, Shota Asada, Atsushi Matsuo, Minako Kishikawa, Kenta Mizuno, Akira Numano, Rika PLoS One Research Article 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 10(3) 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. Public Library of Science 2015-12-09 /pmc/articles/PMC4674099/ /pubmed/26649904 http://dx.doi.org/10.1371/journal.pone.0144254 Text en © 2015 Kurita et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Kurita, Hirofumi
Takahashi, Shota
Asada, Atsushi
Matsuo, Minako
Kishikawa, Kenta
Mizuno, Akira
Numano, Rika
Novel Parallelized Electroporation by Electrostatic Manipulation of a Water-in-Oil Droplet as a Microreactor
title Novel Parallelized Electroporation by Electrostatic Manipulation of a Water-in-Oil Droplet as a Microreactor
title_full Novel Parallelized Electroporation by Electrostatic Manipulation of a Water-in-Oil Droplet as a Microreactor
title_fullStr Novel Parallelized Electroporation by Electrostatic Manipulation of a Water-in-Oil Droplet as a Microreactor
title_full_unstemmed Novel Parallelized Electroporation by Electrostatic Manipulation of a Water-in-Oil Droplet as a Microreactor
title_short Novel Parallelized Electroporation by Electrostatic Manipulation of a Water-in-Oil Droplet as a Microreactor
title_sort novel parallelized electroporation by electrostatic manipulation of a water-in-oil droplet as a microreactor
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4674099/
https://www.ncbi.nlm.nih.gov/pubmed/26649904
http://dx.doi.org/10.1371/journal.pone.0144254
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