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

Scientific Reports
A T Y YeungGordon Dougan

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.

References

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Citations

May 27, 2016·Scandinavian Journal of Immunology·C YangH Du
Dec 13, 2016·Journal of Tissue Engineering and Regenerative Medicine·Viktoriia Starokozhko, Geny M M Groothuis
Apr 20, 2019·Scandinavian Journal of Immunology·Caiyun QianDaya Luo
Feb 27, 2021·Frontiers in Immunology·Clare PridansDavid A Hume

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Datasets Mentioned

BETA
ERP002100

Methods Mentioned

BETA
gene knock out
flow cytometry
RNA-Seq
transfecting
transmission electron microscopy
RNA Seq
gene trapping
scraping
Assay
pull down

Software Mentioned

house
R
InnateDB
Innate
GraphPad Prism
xPONENT Multiplex Assay Analysis
Burrows Aligner
DESeq package

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