TY - JOUR KW - 3D culture KW - Adipose Tissue KW - bone marrow KW - Metabolism KW - Phagocytosis KW - Resident macrophage KW - macrophage subpopulation AU - Adèle Arlat AU - Marie-Laure Renoud AU - Jean Nakhle AU - Miguel Thomas AU - Jessica Fontaine AU - Emmanuelle Arnaud AU - Cédric Dray AU - Hélène Authier AU - Paul Monsarrat AU - Agnès Coste AU - Louis Casteilla AU - Marielle Ousset AU - Béatrice Cousin AB - Introduction

Within adipose tissue (AT), different macrophage subsets have been described, which played pivotal and specific roles in upholding tissue homeostasis under both physiological and pathological conditions. Nonetheless, studying resident macrophages in-vitro poses challenges, as the isolation process and the culture for extended periods can alter their inherent properties.

Methods

Stroma-vascular cells isolated from murine subcutaneous AT were seeded on ultra-low adherent plates in the presence of macrophage colony-stimulating factor. After 4 days of culture, the cells spontaneously aggregate to form spheroids. A week later, macrophages begin to spread out of the spheroid and adhere to the culture plate.

Results

This innovative three-dimensional (3D) culture method enables the generation of functional mature macrophages that present distinct genic and phenotypic characteristics compared to bone marrow–derived macrophages. They also show specific metabolic activity and polarization in response to stimulation, but similar phagocytic capacity. Additionally, based on single-cell analysis, AT-macrophages generated in 3D culture mirror the phenotypic and functional traits of in-vivo AT resident macrophages.

Discussion

Our study describes a 3D in-vitro system for generating and culturing functional AT-resident macrophages, without the need for cell sorting. This system thus stands as a valuable resource for exploring the differentiation and function of AT-macrophages in vitro in diverse physiological and pathological contexts.

 BT - Frontiers in Immunology DA - 2024-06-21 DO - 10.3389/fimmu.2024.1356397 LA - English N2 - Introduction

Within adipose tissue (AT), different macrophage subsets have been described, which played pivotal and specific roles in upholding tissue homeostasis under both physiological and pathological conditions. Nonetheless, studying resident macrophages in-vitro poses challenges, as the isolation process and the culture for extended periods can alter their inherent properties.

Methods

Stroma-vascular cells isolated from murine subcutaneous AT were seeded on ultra-low adherent plates in the presence of macrophage colony-stimulating factor. After 4 days of culture, the cells spontaneously aggregate to form spheroids. A week later, macrophages begin to spread out of the spheroid and adhere to the culture plate.

Results

This innovative three-dimensional (3D) culture method enables the generation of functional mature macrophages that present distinct genic and phenotypic characteristics compared to bone marrow–derived macrophages. They also show specific metabolic activity and polarization in response to stimulation, but similar phagocytic capacity. Additionally, based on single-cell analysis, AT-macrophages generated in 3D culture mirror the phenotypic and functional traits of in-vivo AT resident macrophages.

Discussion

Our study describes a 3D in-vitro system for generating and culturing functional AT-resident macrophages, without the need for cell sorting. This system thus stands as a valuable resource for exploring the differentiation and function of AT-macrophages in vitro in diverse physiological and pathological contexts.

 PY - 2024 T2 - Frontiers in Immunology TI - Generation of functionally active resident macrophages from adipose tissue by 3D cultures UR - https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2024.1356397/full VL - 15 Y2 - 2024-12-30 SN - 1664-3224 ER -