UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
在巨噬细胞集落刺激因子存在下,将从鼠皮下AT分离的基质-血管细胞接种在超低粘附板上。培养4天后,细胞自发聚集形成球体。一周后,巨噬细胞开始从球体中扩散并粘附到培养板上。
■这种创新的三维(3D)培养方法能够产生功能性成熟的巨噬细胞,与骨髓来源的巨噬细胞相比,这些巨噬细胞具有不同的基因和表型特征。它们还显示出特定的代谢活性和对刺激的反应极化,但吞噬能力相似。此外,基于单细胞分析,在3D培养中产生的AT-巨噬细胞反映了体内AT常驻巨噬细胞的表型和功能特征。
■我们的研究描述了一种用于产生和培养功能性AT驻留巨噬细胞的3D体外系统,不需要细胞分选。因此,该系统是探索AT-巨噬细胞在不同生理和病理环境中体外分化和功能的宝贵资源。