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

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, Robert Hancock, G. Dougan

Research output: Contribution to journalArticle

7 Citations (Scopus)

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.

LanguageEnglish
Article number8908
JournalScientific Reports
Volume5
DOIs
Publication statusPublished - 1 Jan 2015

ASJC Scopus subject areas

  • General

Cite this

Yeung, 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, Robert ; Dougan, G. / Conditional-ready mouse embryonic stem cell derived macrophages enable the study of essential genes in macrophage function. In: Scientific Reports. 2015 ; Vol. 5.
@article{e82806f9671c45a59a933009264e337f,
title = "Conditional-ready mouse embryonic stem cell derived macrophages enable the study of essential genes in macrophage function",
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-{\^I}± 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.",
author = "Yeung, {A. T.Y.} and C. Hale and J. Xia and Tate, {P. H.} and D. Goulding and Keane, {J. A.} and S. Mukhopadhyay and L. Forrester and O. Billker and Skarnes, {W. C.} and Robert Hancock and G. Dougan",
year = "2015",
month = "1",
day = "1",
doi = "10.1038/srep08908",
language = "English",
volume = "5",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",

}

Yeung, ATY, Hale, C, Xia, J, Tate, PH, Goulding, D, Keane, JA, Mukhopadhyay, S, Forrester, L, Billker, O, Skarnes, WC, Hancock, R & Dougan, G 2015, 'Conditional-ready mouse embryonic stem cell derived macrophages enable the study of essential genes in macrophage function', Scientific Reports, vol. 5, 8908. https://doi.org/10.1038/srep08908

Conditional-ready mouse embryonic stem cell derived macrophages enable the study of essential genes in macrophage function. / Yeung, 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, Robert; Dougan, G.

In: Scientific Reports, Vol. 5, 8908, 01.01.2015.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Conditional-ready mouse embryonic stem cell derived macrophages enable the study of essential genes in macrophage function

AU - Yeung, A. T.Y.

AU - Hale, C.

AU - Xia, J.

AU - Tate, P. H.

AU - Goulding, D.

AU - Keane, J. A.

AU - Mukhopadhyay, S.

AU - Forrester, L.

AU - Billker, O.

AU - Skarnes, W. C.

AU - Hancock, Robert

AU - Dougan, G.

PY - 2015/1/1

Y1 - 2015/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84924370813&partnerID=8YFLogxK

U2 - 10.1038/srep08908

DO - 10.1038/srep08908

M3 - Article

VL - 5

JO - Scientific Reports

T2 - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 8908

ER -