Abstract

Immune cells resident in adipose tissue have important functions in local and systemic metabolic homeostasis. Nevertheless, these immune cell populations remain poorly characterized in bovines. Recently, we described diverse lymphocyte subpopulations in adipose tissue of Holstein-Friesian cows. Here, we aimed at characterising myeloid cell populations present in bovine adipose tissue using multicolour flow cytometry, cell sorting and histochemistry/immunohistochemistry. Macrophages, CD14+CD11b+MHC-II+CD45+ cells, were identified in mesenteric and subcutaneous adipose tissue, though at higher proportions in the latter. Mast cells, identified as SSC-AhighCD11b−/+CD14−MHC-II−CH138A−CD45+ cells, were also observed in adipose tissue and found at higher proportions than macrophages in mesenteric adipose tissue. Neutrophils, presenting a CH138A+CD11b+ phenotype, were also detected in mesenteric and subcutaneous adipose tissue, however, at much lower frequencies than in the blood. Our gating strategy allowed identification of eosinophils in blood but not in adipose tissue although being detected by morphological analysis at low frequencies in some animals. A population not expressing CD45 and with the CH138A+ CD11b−MHC-II− phenotype, was found abundant and present at higher proportions in mesenteric than subcutaneous adipose tissue. The work reported here may be useful for further studies addressing the function of the described cells.

Highlights

  • Colour-flow cytometry revealed the presence of CD14+ cells, CD172a+ cells, CD11c+ cells and CD163+ cells in the stromal vascular fraction (SVF) of subcutaneous and omental adipose tissue[23,24]

  • We showed that CD45 negative cells represent more than 40% of all the cells in the adipose tissue SVF of mesenteric adipose tissue (MAT)

  • In contrast to blood monocytes, CD11c expression was only observed in a fraction of adipose tissue macrophages (22,4% and 33,05% of total macrophages in MAT and subcutaneous adipose tissue (SAT), respectively) (Fig. 2d)

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Summary

Introduction

Colour-flow cytometry revealed the presence of CD14+ cells, CD172a+ cells, CD11c+ cells and CD163+ cells in the stromal vascular fraction (SVF) of subcutaneous and omental adipose tissue[23,24]. Aylward et al.[25] reported the presence of CD172a+CD11b+cells, CD11c+MHC-II+ cells, and CD11b+CD11c+ cells in mesenteric adipose tissue. These markers are not exclusive to macrophages but common to other myeloid cell populations or can be even expressed in non-hematopoietic cells. To characterize the different myeloid cell populations in bovine adipose tissue we used a multicolour flow cytometry panel associated with cell sorting and histochemistry/immunohistochemistry analysis. Our strategy allowed identification and isolation of macrophages, mast cells and neutrophils with macrophages being observed at different proportions in mesenteric adipose tissue (MAT) and subcutaneous adipose tissue (SAT). Our results highlight that distinct adipose tissue depots may present distinct cellular characteristics

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