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D-Glucosamine Promotes Transfection Efficiency during Electroporation

D-Glucosamine is a useful medicament in various fields of medicine and dentistry. With respect to stability of the cell membrane, it has been reported that bradykinin-induced nociceptive responses are significantly suppressed by the direct application of D-glucosamine. Electroporation is usually use...

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Autores principales: Igawa, Kazunari, Ohara, Naoko, Kawakubo, Atsushi, Sugimoto, Kouji, Yanagiguchi, Kajiro, Ikeda, Takeshi, Yamada, Shizuka, Hayashi, Yoshihiko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3942286/
https://www.ncbi.nlm.nih.gov/pubmed/24678506
http://dx.doi.org/10.1155/2014/485867
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author Igawa, Kazunari
Ohara, Naoko
Kawakubo, Atsushi
Sugimoto, Kouji
Yanagiguchi, Kajiro
Ikeda, Takeshi
Yamada, Shizuka
Hayashi, Yoshihiko
author_facet Igawa, Kazunari
Ohara, Naoko
Kawakubo, Atsushi
Sugimoto, Kouji
Yanagiguchi, Kajiro
Ikeda, Takeshi
Yamada, Shizuka
Hayashi, Yoshihiko
author_sort Igawa, Kazunari
collection PubMed
description D-Glucosamine is a useful medicament in various fields of medicine and dentistry. With respect to stability of the cell membrane, it has been reported that bradykinin-induced nociceptive responses are significantly suppressed by the direct application of D-glucosamine. Electroporation is usually used to effectively introduce foreign genes into tissue culture cells. Buffers for electroporation with or without D-glucosamine are used in experiments of transfection vectors. This is the first study to indirectly observe the stability and protection of the osteoblast membrane against both electric stress and gene uptake (the proton sponge hypothesis: osmotic rupture during endosomes prior to fusion with lysosomes) in electroporation with D-glucosamine application. The transfection efficiency was evaluated as the fluorescence intensity of the transfected green fluorescent protein (GFP) in the cultured cells (osteoblasts; NOS-1 cells). The transfection efficiency increased over 30% in the electroporation samples treated with D-glucosamine-supplemented buffer after one day. The membrane absorption of D-glucosamine is the primary mechanism of membrane stress induced by electric stress. This new function of D-glucosamine is useful and meaningful for developing more effective transformation procedures.
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spelling pubmed-39422862014-03-27 D-Glucosamine Promotes Transfection Efficiency during Electroporation Igawa, Kazunari Ohara, Naoko Kawakubo, Atsushi Sugimoto, Kouji Yanagiguchi, Kajiro Ikeda, Takeshi Yamada, Shizuka Hayashi, Yoshihiko Biomed Res Int Research Article D-Glucosamine is a useful medicament in various fields of medicine and dentistry. With respect to stability of the cell membrane, it has been reported that bradykinin-induced nociceptive responses are significantly suppressed by the direct application of D-glucosamine. Electroporation is usually used to effectively introduce foreign genes into tissue culture cells. Buffers for electroporation with or without D-glucosamine are used in experiments of transfection vectors. This is the first study to indirectly observe the stability and protection of the osteoblast membrane against both electric stress and gene uptake (the proton sponge hypothesis: osmotic rupture during endosomes prior to fusion with lysosomes) in electroporation with D-glucosamine application. The transfection efficiency was evaluated as the fluorescence intensity of the transfected green fluorescent protein (GFP) in the cultured cells (osteoblasts; NOS-1 cells). The transfection efficiency increased over 30% in the electroporation samples treated with D-glucosamine-supplemented buffer after one day. The membrane absorption of D-glucosamine is the primary mechanism of membrane stress induced by electric stress. This new function of D-glucosamine is useful and meaningful for developing more effective transformation procedures. Hindawi Publishing Corporation 2014 2014-02-11 /pmc/articles/PMC3942286/ /pubmed/24678506 http://dx.doi.org/10.1155/2014/485867 Text en Copyright © 2014 Kazunari Igawa et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Igawa, Kazunari
Ohara, Naoko
Kawakubo, Atsushi
Sugimoto, Kouji
Yanagiguchi, Kajiro
Ikeda, Takeshi
Yamada, Shizuka
Hayashi, Yoshihiko
D-Glucosamine Promotes Transfection Efficiency during Electroporation
title D-Glucosamine Promotes Transfection Efficiency during Electroporation
title_full D-Glucosamine Promotes Transfection Efficiency during Electroporation
title_fullStr D-Glucosamine Promotes Transfection Efficiency during Electroporation
title_full_unstemmed D-Glucosamine Promotes Transfection Efficiency during Electroporation
title_short D-Glucosamine Promotes Transfection Efficiency during Electroporation
title_sort d-glucosamine promotes transfection efficiency during electroporation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3942286/
https://www.ncbi.nlm.nih.gov/pubmed/24678506
http://dx.doi.org/10.1155/2014/485867
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