Pleura organoid transplantation facilitates visceral pleural repair through CD133+ progenitor cell-mediated regeneration.

Persistent pulmonary air leakage following thoracic surgery remains a major clinical challenge. Here we establish murine and human pleura organoids as a novel regenerative therapy. Morphological and functional analyses confirm that these organoids retain mesothelial markers and secretory activity critical for pleural homeostasis. Xenotransplantation beneath the renal capsule of immunodeficient mice generates pleura-like structures with a mesothelial monolayer and collagen-elastin networks. In vitro wound healing and precision-cut lung slice models demonstrate enhanced mesothelial migration and organoid integration at injury sites. In vivo, transplantation into a murine pleural rupture model facilitates dynamic structural restoration, with continuous mesothelial layer reformation and sustained recovery of pleural barrier function, and attenuates acute inflammation and fibrosis. Transcriptomic and functional studies, notably human lung single-cell RNA sequencing analysis, identify CD133+ cells as a key subpopulation, exhibiting enhanced organoid-forming capacity and multilineage differentiation in vitro. Critically, CD133+ organoid transplantation significantly alleviates murine pleural rupture. This pioneers a biologically precise interventions, elucidating the important role of CD133+ cells in pleura repair, with transformative potential for thoracic regenerative medicine.