A bioengineered intestinal epithelial-stromal model for studying coordinated epithelial migration during wound repair

Fibroblasts reside underneath most epithelial tissues. In the intestine, recent studies have shown that fibroblast migration contributes to tissue morphogenesis and wound healing. Yet, whether physical interactions between epithelial cells and fibroblasts contribute to epithelial movement remains elusive. Here, we establish a reconstituted epithelial-stromal gap-closure model that enables the study of physical epithelial-fibroblast interactions during intestinal repair. Using this platform, we show that subepithelial fibroblast alignment enhances directed and persistent migration of organoid-derived intestinal epithelia. We demonstrate that direct contact with fibroblasts improves gap closure by promoting cell alignment, sustaining tissue integrity, and synchronizing crypt-villus migration. Fibroblasts undergo long-range ordering to align perpendicularly to the epithelial front and deposit protein paths that act as guidance features to direct epithelial migration. In parallel, epithelial cells acquire a wound-associated epithelial-like phenotype, but insufficient to explain the effects of fibroblast contact. Our findings uncover a dual role for intestinal fibroblasts in epithelial repair, coordinating both biochemical and physical cues to ensure efficient and cohesive migration.