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Chicken CSF2 and IL-4-, and CSF2-dependent bone marrow cultures differentiate into macrophages over time

Chicken bone marrow-derived macrophages (BMMΦ) and dendritic cells (BMDC) are utilized as models to study the mononuclear phagocytic system (MPS). A widely used method to generate macrophages and DC in vitro is to culture bone marrow cells in the presence of colony-stimulating factor-1 (CSF1) to dif...

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Autores principales: Borowska, Dominika, Sives, Samantha, Vervelde, Lonneke, Sutton, Kate M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9812659/
https://www.ncbi.nlm.nih.gov/pubmed/36618373
http://dx.doi.org/10.3389/fimmu.2022.1064084
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author Borowska, Dominika
Sives, Samantha
Vervelde, Lonneke
Sutton, Kate M.
author_facet Borowska, Dominika
Sives, Samantha
Vervelde, Lonneke
Sutton, Kate M.
author_sort Borowska, Dominika
collection PubMed
description Chicken bone marrow-derived macrophages (BMMΦ) and dendritic cells (BMDC) are utilized as models to study the mononuclear phagocytic system (MPS). A widely used method to generate macrophages and DC in vitro is to culture bone marrow cells in the presence of colony-stimulating factor-1 (CSF1) to differentiate BMMΦ and granulocyte-macrophage-CSF (GM-CSF, CSF2) and interleukin-4 (IL-4) to differentiate BMDC, while CSF2 alone can lead to the development of granulocyte-macrophage-CSF-derived DC (GMDC). However, in chickens, the MPS cell lineages and their functions represented by these cultures are poorly understood. Here, we decipher the phenotypical, functional and transcriptional differences between chicken BMMΦ and BMDC along with examining differences in DC cultures grown in the absence of IL-4 on days 2, 4, 6 and 8 of culture. BMMΦ cultures develop into a morphologically homogenous cell population in contrast to the BMDC and GMDC cultures, which produce morphologically heterogeneous cell cultures. At a phenotypical level, all cultures contained similar cell percentages and expression levels of MHCII, CD11c and CSF1R-transgene, whilst MRC1L-B expression decreased over time in BMMΦ. All cultures were efficiently able to uptake 0.5 µm beads, but poorly phagocytosed 1 µm beads. Little difference was observed in the kinetics of phagosomal acidification across the cultures on each day of analysis. Temporal transcriptomic analysis indicated that all cultures expressed high levels of CSF3R, MERTK, SEPP1, SPI1 and TLR4, genes associated with macrophages in mammals. In contrast, low levels of FLT3, XCR1 and CAMD1, genes associated with DC, were expressed at day 2 in BMDC and GMDC after which expression levels decreased. Collectively, chicken CSF2 + IL-4- and CSF2-dependent BM cultures represent cells of the macrophage lineage rather than inducing conventional DC.
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spelling pubmed-98126592023-01-05 Chicken CSF2 and IL-4-, and CSF2-dependent bone marrow cultures differentiate into macrophages over time Borowska, Dominika Sives, Samantha Vervelde, Lonneke Sutton, Kate M. Front Immunol Immunology Chicken bone marrow-derived macrophages (BMMΦ) and dendritic cells (BMDC) are utilized as models to study the mononuclear phagocytic system (MPS). A widely used method to generate macrophages and DC in vitro is to culture bone marrow cells in the presence of colony-stimulating factor-1 (CSF1) to differentiate BMMΦ and granulocyte-macrophage-CSF (GM-CSF, CSF2) and interleukin-4 (IL-4) to differentiate BMDC, while CSF2 alone can lead to the development of granulocyte-macrophage-CSF-derived DC (GMDC). However, in chickens, the MPS cell lineages and their functions represented by these cultures are poorly understood. Here, we decipher the phenotypical, functional and transcriptional differences between chicken BMMΦ and BMDC along with examining differences in DC cultures grown in the absence of IL-4 on days 2, 4, 6 and 8 of culture. BMMΦ cultures develop into a morphologically homogenous cell population in contrast to the BMDC and GMDC cultures, which produce morphologically heterogeneous cell cultures. At a phenotypical level, all cultures contained similar cell percentages and expression levels of MHCII, CD11c and CSF1R-transgene, whilst MRC1L-B expression decreased over time in BMMΦ. All cultures were efficiently able to uptake 0.5 µm beads, but poorly phagocytosed 1 µm beads. Little difference was observed in the kinetics of phagosomal acidification across the cultures on each day of analysis. Temporal transcriptomic analysis indicated that all cultures expressed high levels of CSF3R, MERTK, SEPP1, SPI1 and TLR4, genes associated with macrophages in mammals. In contrast, low levels of FLT3, XCR1 and CAMD1, genes associated with DC, were expressed at day 2 in BMDC and GMDC after which expression levels decreased. Collectively, chicken CSF2 + IL-4- and CSF2-dependent BM cultures represent cells of the macrophage lineage rather than inducing conventional DC. Frontiers Media S.A. 2022-12-21 /pmc/articles/PMC9812659/ /pubmed/36618373 http://dx.doi.org/10.3389/fimmu.2022.1064084 Text en Copyright © 2022 Borowska, Sives, Vervelde and Sutton https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Borowska, Dominika
Sives, Samantha
Vervelde, Lonneke
Sutton, Kate M.
Chicken CSF2 and IL-4-, and CSF2-dependent bone marrow cultures differentiate into macrophages over time
title Chicken CSF2 and IL-4-, and CSF2-dependent bone marrow cultures differentiate into macrophages over time
title_full Chicken CSF2 and IL-4-, and CSF2-dependent bone marrow cultures differentiate into macrophages over time
title_fullStr Chicken CSF2 and IL-4-, and CSF2-dependent bone marrow cultures differentiate into macrophages over time
title_full_unstemmed Chicken CSF2 and IL-4-, and CSF2-dependent bone marrow cultures differentiate into macrophages over time
title_short Chicken CSF2 and IL-4-, and CSF2-dependent bone marrow cultures differentiate into macrophages over time
title_sort chicken csf2 and il-4-, and csf2-dependent bone marrow cultures differentiate into macrophages over time
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9812659/
https://www.ncbi.nlm.nih.gov/pubmed/36618373
http://dx.doi.org/10.3389/fimmu.2022.1064084
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