Journal of Capital Medical University ›› 2026, Vol. 47 ›› Issue (4): 707-714.doi: 10.3969/j.issn.1006-7795.2026.04.011

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Establishment of nasal mucosal epithelial organoids and assessment of their barrier function

Wang Jingwen1,2,3, Zhang Yuan3,4, Zhang Xu1,2,3 , Zhang Luo1,2,3,4*, Li Jingyun1,2,3*   

  1. 1.Department of Otolaryngology Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China; 2.Beijing Laboratory of Allergic Diseases, Beijing Municipal Education Commission and Beijing Key Laboratory of New Medicine and Diagnostic Technology Research for Nasal Disease, Beijing Institute of Otolaryngology, Beijing 100005, China； 3.National Engineering Research Center for Innovative Drugs for Allergic Diseases, Beijing Tongren Hospital, Capital Medical University, Beijing 100005, China; 4.Department of Allergy, Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China
  • Received:2026-04-09 Revised:2026-06-09 Online:2026-08-21 Published:2026-07-26
  • Supported by:
    This study was supported by National Natural Science Foundation of China (81970849,82271141).

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Abstract: Objective  To establish, characterize and validate nasal epithelial organoids derived from surgical specimens of patients with chronic rhinosinusitis with nasal polyps (CRSwNP) and to comprehensively evaluate their growth, cellular composition, and functional epithelial barrier properties. Methods  Cell clusters isolated from CRSwNP specimens were encapsulated in 3D Matrigel and serially passaged. Organoid growth was quantified by organoid quantity, average area of organoids, and organoid forming efficiency (OFE). Cellular composition was assessed using flow cytometry, immunohistochemistry(IHC), and immunofluorescence(IF) for canonical epithelial markers. Barrier function was evaluated by transepithelial electrical resistance (TEER). Results  CRSwNP-derived clusters developed into compact, well-defined 3D organoids that remained at least five generation (P5). At day 7 post-seeding (D7)—a time point representing structural maturation—organoid quantity and OFE increased significantly compared with day 1 (D1) (P < 0.05), whereas organoid area remained statistically invariant (P > 0.05), indicating preserved clonogenic efficiency and proliferative fidelity across passages.  KRT5-positive basal cells accounted for >90% of undifferentiated organoids with stable distribution across passages. TEER increased progressively during differentiation, indicating establishment of epithelial barrier function. Differentiated organoids expressed β-tubulin (ciliated cells) and MUC5AC (goblet cells). Conclusion  We  established patient-derived, CRSwNP-origin nasal epithelial organoids that retain key structural, cellular, and functional features. Organoid quantity, area, and OFE are effective indicators for quantifying growth, and combined with TEER and differentiation markers, they are used to evaluated barrier function. This model offers a robust in vitro platform for investigating nasal inflammatory disease mechanisms and for the preclinical evaluation of novel therapeutics, including biologics.

Key words: chronic rhinosinusitis with nasal polyps, nasal epithelial organoids, basal cells, cell differentiation, epithelial barrier function, three-dimensional culture 

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