Conditional-ready mouse embryonic stem cell derived macrophages enable the study of essential genes in macrophage function.
AuthorYeung, ATY; Hale, C; Xia, J; Tate, PH; Goulding, D; Keane, JA; Mukhopadhyay, S; Forrester, L; Billker, O; Skarnes, WC; ...
Source TitleScientific Reports
PublisherSpringer Science and Business Media LLC
University of Melbourne Author/sDougan, Gordon
AffiliationMicrobiology and Immunology
Document TypeJournal Article
CitationsYeung, A. T. Y., Hale, C., Xia, J., Tate, P. H., Goulding, D., Keane, J. A., Mukhopadhyay, S., Forrester, L., Billker, O., Skarnes, W. C., Hancock, R. E. W. & Dougan, G. (2015). Conditional-ready mouse embryonic stem cell derived macrophages enable the study of essential genes in macrophage function.. Sci Rep, 5 (1), pp.8908-. https://doi.org/10.1038/srep08908.
Access StatusOpen Access
Open Access at PMChttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC4354151
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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