title: A cell line resource derived from honey bee (Apis mellifera) embryonic tissues.
abstract: A major hindrance to the study of honey bee pathogens or the effects of pesticides and nutritional deficiencies is the lack of controlled in vitro culture systems comprised of honey bee cells. Such systems are important to determine the impact of these stress factors on the developmental and cell biology of honey bees. We have developed a method incorporating established insect cell culture techniques that supports sustained growth of honey bee cells in vitro. We used honey bee eggs mid to late in their embryogenesis to establish primary cultures, as these eggs contain cells that are progressively dividing. Primary cultures were initiated in modified Leibovitz's L15 medium and incubated at 32(°)C. Serial transfer of material from several primary cultures was maintained and has led to the isolation of young cell lines. A cell line (AmE-711) has been established that is composed mainly of fibroblast-type cells that form an adherent monolayer. Most cells in the line are diploid (2n = 32) and have the Apis mellifera karyotype as revealed by Giemsa stain. The partial sequence for the mitochondrial-encoded cytochrome c oxidase subunit I (Cox 1) gene in the cell line is identical to those from honey bee tissues and a consensus sequence for A. mellifera. The population doubling time is approximately 4 days. Importantly, the cell line is continuously subcultured every 10-14 days when split at a 1:3 ratio and is cryopreserved in liquid nitrogen. The cell culture system we have developed has potential application for studies aimed at honey bee development, genetics, pathogenesis, transgenesis, and toxicology.
authors.affiliation: Department of Entomology, University of Minnesota, St. Paul, Minnesota, USA. goblirmj@umn.edu
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Goblirsch
authors.forename: Marla S
authors.initials: MS
authors.lastname: Spivak
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Kurtti
mesh_terms: D000818:Animals; D001516:Bees / Q000166:cytology*; D002460:Cell Line / Q000201:enzymology; D004171:Diploidy; D003576:Electron Transport Complex IV / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000166:cytology; D021122:Protein Subunits / Q000378:metabolism
chemical_list: D021122:Protein Subunits; D003576:Electron Transport Complex IV
references.citation: J Insect Sci. 2002;2:9
references.pmid: 15455043
references.citation: Environ Microbiol. 2011 Feb;13(2):404-13
references.pmid: 20880328
references.citation: Tissue Cell. 1994 Aug;26(4):551-8
references.pmid: 18621280
references.citation: C R Acad Hebd Seances Acad Sci D. 1967 Sep 11;265(11):800-3
references.pmid: 4965090
references.citation: Nature. 2006 Oct 26;443(7114):931-49
references.pmid: 17073008
references.citation: J Gen Virol. 2004 Apr;85(Pt 4):863-868
references.pmid: 15039529
references.citation: In Vitro Cell Dev Biol Anim. 2011 Aug;47(7):451-3
references.pmid: 21688083
references.citation: J Invertebr Pathol. 2009 Nov;102(3):256-62
references.pmid: 19761771
references.citation: Exp Appl Acarol. 1989 Aug;7(3):219-29
references.pmid: 2766897
references.citation: Nature. 1962 Aug 25;195:788-9
references.pmid: 13900940
references.citation: Am J Hyg. 1963 Sep;78:173-80
references.pmid: 14063721
references.citation: In Vitro Cell Dev Biol Anim. 2010 Feb;46(2):83-6
references.pmid: 20033792
references.citation: Proc Natl Acad Sci U S A. 1991 May 15;88(10):4548-52
references.pmid: 1674608
references.citation: J Clin Microbiol. 1999 Aug;37(8):2518-24
references.pmid: 10405394
references.citation: Annu Rev Entomol. 1971;16:27-52
references.pmid: 4930372
references.citation: PLoS Pathog. 2012;8(5):e1002716
references.pmid: 22615573
references.citation: J Invertebr Pathol. 2013 Mar;112 Suppl:S138-41
references.pmid: 22841637
references.citation: BMC Dev Biol. 2006 Mar 17;6:17
references.pmid: 16545111
references.citation: J Biomed Sci. 2010 May 11;17:36
references.pmid: 20459778
references.citation: In Vitro Cell Dev Biol Anim. 2001 Jun;37(6):319-21
references.pmid: 11515961
references.citation: PLoS One. 2011;6(6):e20656
references.pmid: 21687739
references.citation: Mol Biol Evol. 1989 Jul;6(4):399-411
references.pmid: 2559293
references.citation: Biotechnol Adv. 2012 May-Jun;30(3):766-81
references.pmid: 22297133
references.citation: Insect Mol Biol. 2010 Oct;19(5):653-8
references.pmid: 20546039
references.citation: Science. 2007 Oct 12;318(5848):283-7
references.pmid: 17823314
references.citation: J Invertebr Pathol. 2007 Mar;94(3):211-7
references.pmid: 17217954
references.citation: Invert Neurosci. 2008 Mar;8(1):19-29
references.pmid: 18004599
references.citation: Insect Biochem Mol Biol. 2003 Nov;33(11):1123-34
references.pmid: 14563363
references.citation: J Exp Biol. 1999 Apr;202(Pt 8):929-38
references.pmid: 10085265
references.citation: Genetics. 1993 Jan;133(1):97-117
references.pmid: 8417993
references.citation: J Neurocytol. 1992 Aug;21(8):545-56
references.pmid: 1354720
references.citation: Adv Virus Res. 2007;70:33-80
references.pmid: 17765703
references.citation: Exp Cell Res. 1961 Dec;25:585-621
references.pmid: 13905658
references.citation: Arch Insect Biochem Physiol. 1991;18(2):99-104
references.pmid: 1799679