A macrophage–intestinal organoid co-culture model captures balanced epithelial homeostasis and inflammation
Immune–epithelial interactions are essential for intestinal homeostasis, yet organoid monocultures lack immune components and fail to model how immune cells shape epithelial identity. Here, we establish a macrophage–organoid co-culture integrating bone marrow–derived macrophages (BMDMs) into the intestinal niche to create a controlled, immune-competent system. Under homeostatic conditions, the optimized BMDMs 5k configuration preserves crypt-like architecture, maintains stem cell localization, and secretory lineage organization. TNFa stimulation induces coordinated epithelial remodeling characterized by reduced canonical stem cell marker expression, and density-dependent shifts in macrophage phenotype. Transcriptomic profiling confirms enrichment of immune-associated and epithelial stress pathways. Comparative analysis with a DSS-induced murine inflammation model reveals overlapping inflammatory and epithelial remodeling features, supporting physiological relevance. This platform provides a reproducible framework for studying epithelial–immune interactions and inflammatory remodeling under defined conditions, offering a controlled alternative to animal models.
Immune–epithelial interactions are essential for intestinal homeostasis, yet organoid monocultures lack immune components and fail to model how immune cells shape epithelial identity. Here, we establish a macrophage–organoid co-culture integrating bone marrow–derived macrophages (BMDMs) into the intestinal niche to create a controlled, immune-competent system. Under homeostatic conditions, the optimized BMDMs 5k configuration preserves crypt-like architecture, maintains stem cell localization, and secretory lineage organization. TNFa stimulation induces coordinated epithelial remodeling characterized by reduced canonical stem cell marker expression, and density-dependent shifts in macrophage phenotype. Transcriptomic profiling confirms enrichment of immune-associated and epithelial stress pathways. Comparative analysis with a DSS-induced murine inflammation model reveals overlapping inflammatory and epithelial remodeling features, supporting physiological relevance. This platform provides a reproducible framework for studying epithelial–immune interactions and inflammatory remodeling under defined conditions, offering a controlled alternative to animal models.
Jeju National University