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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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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. |
format | Online Article Text |
id | pubmed-4674099 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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