Format

Send to

Choose Destination
Sci Rep. 2015 Mar 10;5:8908. doi: 10.1038/srep08908.

Conditional-ready mouse embryonic stem cell derived macrophages enable the study of essential genes in macrophage function.

Author information

1
Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, United Kingdom.
2
Centre for Microbial Diseases and Immunity Research, University of British Columbia, Vancouver, BC, Canada.
3
University of Edinburgh/MRC Centre for Regenerative Medicine, Edinburgh, United Kingdom.
4
1] Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, United Kingdom [2] Centre for Microbial Diseases and Immunity Research, University of British Columbia, Vancouver, BC, Canada.

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.

PMID:
25752829
PMCID:
PMC4354151
DOI:
10.1038/srep08908
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center