In:
PLOS ONE, Public Library of Science (PLoS), Vol. 16, No. 9 ( 2021-9-23), p. e0257770-
Abstract:
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.
Type of Medium:
Online Resource
ISSN:
1932-6203
DOI:
10.1371/journal.pone.0257770
DOI:
10.1371/journal.pone.0257770.g001
DOI:
10.1371/journal.pone.0257770.g002
DOI:
10.1371/journal.pone.0257770.g003
DOI:
10.1371/journal.pone.0257770.g004
DOI:
10.1371/journal.pone.0257770.g005
DOI:
10.1371/journal.pone.0257770.g006
DOI:
10.1371/journal.pone.0257770.g007
DOI:
10.1371/journal.pone.0257770.g008
DOI:
10.1371/journal.pone.0257770.g009
DOI:
10.1371/journal.pone.0257770.g010
DOI:
10.1371/journal.pone.0257770.t001
DOI:
10.1371/journal.pone.0257770.s001
DOI:
10.1371/journal.pone.0257770.s002
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2021
detail.hit.zdb_id:
2267670-3
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