A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis.

Cultured cells are a very powerful tool for investigating biological events in vitro; therefore, cell lines have been established not only in model insect species, but also in non-model species. However, there are few reports on the establishment of stable cell lines and development of systems to in...

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Autores principales:	Kazuyo Watanabe, Mikio Yoshiyama, Gaku Akiduki, Kakeru Yokoi, Hiroko Hoshida, Takumi Kayukawa, Kiyoshi Kimura, Masatsugu Hatakeyama
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e50aca0c4083429e8efa6d6dd6be5e39
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spelling	oai:doaj.org-article:e50aca0c4083429e8efa6d6dd6be5e392021-12-02T20:14:12ZA simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis.1932-620310.1371/journal.pone.0257770https://doaj.org/article/e50aca0c4083429e8efa6d6dd6be5e392021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0257770https://doaj.org/toc/1932-6203Cultured cells are a very powerful tool for investigating biological events in vitro; therefore, cell lines have been established not only in model insect species, but also in non-model species. However, there are few reports on the establishment of stable cell lines and development of systems to introduce genes into the cultured cells of the honey bee (Apis mellifera). We describe a simple ex vivo cell culture system for the honey bee. Hemocyte cells obtained from third and fourth instar larvae were cultured in commercial Grace's insect medium or MGM-450 insect medium for more than two weeks maintaining a normal morphology without deterioration. After an expression plasmid vector bearing the enhanced green fluorescent protein (egfp) gene driven by the immediate early 2 (IE2) viral promoter was transfected into cells, EGFP fluorescence was detected in cells for more than one week from one day after transfection. Furthermore, double-stranded RNA corresponding to a part of the egfp gene was successfully introduced into cells and interfered with egfp gene expression. A convenient and reproducible method for an ex vivo cell culture that is fully practicable for gene expression assays was established for the honey bee.Kazuyo WatanabeMikio YoshiyamaGaku AkidukiKakeru YokoiHiroko HoshidaTakumi KayukawaKiyoshi KimuraMasatsugu HatakeyamaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 9, p e0257770 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kazuyo Watanabe Mikio Yoshiyama Gaku Akiduki Kakeru Yokoi Hiroko Hoshida Takumi Kayukawa Kiyoshi Kimura Masatsugu Hatakeyama A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis.
description	Cultured cells are a very powerful tool for investigating biological events in vitro; therefore, cell lines have been established not only in model insect species, but also in non-model species. However, there are few reports on the establishment of stable cell lines and development of systems to introduce genes into the cultured cells of the honey bee (Apis mellifera). We describe a simple ex vivo cell culture system for the honey bee. Hemocyte cells obtained from third and fourth instar larvae were cultured in commercial Grace's insect medium or MGM-450 insect medium for more than two weeks maintaining a normal morphology without deterioration. After an expression plasmid vector bearing the enhanced green fluorescent protein (egfp) gene driven by the immediate early 2 (IE2) viral promoter was transfected into cells, EGFP fluorescence was detected in cells for more than one week from one day after transfection. Furthermore, double-stranded RNA corresponding to a part of the egfp gene was successfully introduced into cells and interfered with egfp gene expression. A convenient and reproducible method for an ex vivo cell culture that is fully practicable for gene expression assays was established for the honey bee.
format	article
author	Kazuyo Watanabe Mikio Yoshiyama Gaku Akiduki Kakeru Yokoi Hiroko Hoshida Takumi Kayukawa Kiyoshi Kimura Masatsugu Hatakeyama
author_facet	Kazuyo Watanabe Mikio Yoshiyama Gaku Akiduki Kakeru Yokoi Hiroko Hoshida Takumi Kayukawa Kiyoshi Kimura Masatsugu Hatakeyama
author_sort	Kazuyo Watanabe
title	A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis.
title_short	A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis.
title_full	A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis.
title_fullStr	A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis.
title_full_unstemmed	A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis.
title_sort	simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/e50aca0c4083429e8efa6d6dd6be5e39
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A simple method for ex vivo honey bee cell culture capable of in vitro gene expression analysis.

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