Abstract |
The ability to differentiate genetically modified mouse embryonic stem (ES) cells into functional macrophages provides a potentially attractive resource to study host-pathogen interactions without the need for animal experimentation. This is particularly useful in instances where the gene of interest is essential and a knockout mouse is not available. Here we differentiated mouse ES cells into macrophages in vitro and showed, through a combination of flow cytometry, microscopic imaging, and RNA-Seq, that ES cell-derived macrophages responded to S. Typhimurium, in a comparable manner to mouse bone marrow derived macrophages. We constructed a homozygous mutant mouse ES cell line in the Traf2 gene that is known to play a role in tumour necrosis factor-α signalling but has not been studied for its role in infections or response to Toll-like receptor agonists. Interestingly, traf2-deficient macrophages produced reduced levels of inflammatory cytokines in response to lipopolysaccharide (LPS) or flagellin stimulation and exhibited increased susceptibility to S. Typhimurium infection.
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Authors | A T Y Yeung, C Hale, J Xia, P H Tate, D Goulding, J A Keane, S Mukhopadhyay, L Forrester, O Billker, W C Skarnes, R E W Hancock, G Dougan |
Journal | Scientific reports
(Sci Rep)
Vol. 5
Pg. 8908
(Mar 10 2015)
ISSN: 2045-2322 [Electronic] England |
PMID | 25752829
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Lipopolysaccharides
- TNF Receptor-Associated Factor 2
- Toll-Like Receptors
- Tumor Necrosis Factor-alpha
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Topics |
- Animals
- Cell Differentiation
(genetics)
- Gene Expression Regulation, Developmental
(drug effects, genetics)
- Humans
- Lipopolysaccharides
(toxicity)
- Macrophages
(drug effects, metabolism)
- Mice
- Mouse Embryonic Stem Cells
(cytology, metabolism)
- Salmonella typhimurium
(pathogenicity)
- Signal Transduction
(drug effects, genetics)
- TNF Receptor-Associated Factor 2
(biosynthesis, genetics)
- Toll-Like Receptors
(metabolism)
- Tumor Necrosis Factor-alpha
(genetics)
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