02782nas a2200373   4500000000100000000000100001008004100002260001500043653001500058653001900073653001600092653001500108653001700123653002400140653002900164100001700193700002300210700001600233700001800249700002100267700002200288700001700310700002100327700001900348700001700367700002000384700002000404700002100424245009400445856009300539490000700632520175500639022001402394       2024                            d  c2024-06-2110a3D culture10aAdipose Tissue10abone marrow10aMetabolism10aPhagocytosis10aResident macrophage10amacrophage subpopulation1 aAdèle Arlat1 aMarie-Laure Renoud1 aJean Nakhle1 aMiguel Thomas1 aJessica Fontaine1 aEmmanuelle Arnaud1 aCédric Dray1 aHélène Authier1 aPaul Monsarrat1 aAgnès Coste1 aLouis Casteilla1 aMarielle Ousset1 aBéatrice Cousin00aGeneration of functionally active resident macrophages from adipose tissue by 3D cultures  uhttps://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2024.1356397/full0 v153 a<sec><title>Introduction</title><p>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 <italic>in-vitro</italic> poses challenges, as the isolation process and the culture for extended periods can alter their inherent properties.</p></sec><sec><title>Methods</title><p>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.</p></sec><sec><title>Results</title><p>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 <italic>in-vivo</italic> AT resident macrophages.</p></sec><sec><title>Discussion</title><p>Our study describes a 3D <italic>in-vitro</italic> 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 <italic>in vitro</italic> in diverse physiological and pathological contexts.</p></sec>  a1664-3224