Pleura Organoid Transplantation Facilitates Visceral Pleural Repair through CD133+ Progenitor Cell-Mediated Regeneration

Persistent pulmonary air leakage following thoracic surgery remains a significant clinical challenge due to inadequate repair strategies. 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, closely resembling native tissue architecture. 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 restores structural continuity and reduces inflammatory cytokines (Tumor necrosis factor-alpha and Interleukin-6). Transcriptomic and functional studies identify CD133+ cells as a key regenerative subpopulation, exhibiting multilineage differentiation and an enhanced organoid-forming capacity. Transplantation of CD133+ organoids rather than CD133- organoids significantly alleviates murine pleural rupture. This work not only pioneers a biologically precise alternative to current interventions but also reveals CD133+ progenitors as central mediators of mesothelial repair, offering transformative potential for thoracic regenerative medicine.