鼻黏膜上皮类器官的构建及屏障功能分析

doi:10.3969/j.issn.1006-7795.2026.04.011

首都医科大学学报 ›› 2026, Vol. 47 ›› Issue (4): 707-714.doi: 10.3969/j.issn.1006-7795.2026.04.011

鼻黏膜上皮类器官的构建及屏障功能分析

王瀞雯^1,2,3,张媛^3,4,张旭^1,2,3,张罗^1,2,3,4*,李景云^1,2,3*

1.首都医科大学附属北京同仁医院耳鼻咽喉头颈外科，北京 100730；2.北京市耳鼻咽喉科研究所，过敏性疾病北京实验室（北京市教育委员会），慢性鼻病新药及诊断技术研发北京市重点实验室，北京 100005；3.首都医科大学附属北京同仁医院过敏性疾病创新药物国家工程研究中心，北京 100005；4.首都医科大学附属北京同仁医院变态反应（鼻过敏）科，北京 100730

收稿日期:2026-04-09 修回日期:2026-06-09 出版日期:2026-08-21 发布日期:2026-07-26
通讯作者: 张罗, 李景云 E-mail:dr.luozhang@139.com； lijingyun_1234@163.com
基金资助:
国家自然科学基金项目（81970849，82271141）。

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

Wang Jingwen^1,2,3, Zhang Yuan^3,4, Zhang Xu^1,2,3 , Zhang Luo^1,2,3,4*, Li Jingyun^1,2,3*

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).

摘要/Abstract

摘要： 目的探讨利用慢性鼻窦炎伴鼻息肉（chronic rhinosinusitis with nasal polyps, CRSwNP）手术样本来源鼻黏膜上皮类器官的构建及传代可行性，并系统评估生长特性、细胞组成及屏障功能。方法取CRSwNP手术标本分离细胞团，在基质胶中进行三维培养并传代。使用类器官形成数量、类器官平均面积及类器官形成效率（organoid forming efficiency,OFE）量化生长特性。采用流式细胞术、免疫组化和免疫荧光检测细胞标志物，并结合上皮电阻测定（transepithelial electrical resistance,TEER）评估屏障功能。结果 CRSwNP息肉组织细胞团在基质胶条件下成功形成边界清晰、结构致密的三维类器官，并可稳定传代至第五代（P5）以上。下一次传代前类器官形成成熟阶段（Day 7,D7）与传代初始阶段（Day 1,D1）相比，成熟类器官数量和OFE显著增加（P＜0.05），而平均面积无显著变化（P＞0.05），提示类器官在连续传代过程中维持较好的形成能力和增殖潜力。流式细胞术及免疫组化显示,未分化类器官KRT5（基底细胞）占比超过90%，分代间分布稳定。TEER随分化时间显著升高，提示上皮屏障功能逐步建立，分化后类器官呈现β-tubulin（纤毛细胞）和MUC5AC（杯状细胞）标志表达。结论成功构建CRSwNP手术标本来源的可传代鼻上皮类器官。类器官数量、平均面积和OFE为量化生长的有效指标，可结合TEER和分化标记用于评估屏障功能。该模型为鼻腔炎症疾病机制研究及新药（尤其是生物制剂）开发提供了可行的体外实验平台。

关键词: 慢性鼻窦炎伴鼻息肉, 鼻上皮类器官, 基底细胞, 细胞分化, 上皮屏障功能, 三维培养

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

中图分类号:

R765.4+1

王瀞雯, 张媛, 张旭, 张罗, 李景云. 鼻黏膜上皮类器官的构建及屏障功能分析[J]. 首都医科大学学报, 2026, 47(4): 707-714.

Wang Jingwen, Zhang Yuan, Zhang Xu, , Zhang Luo, Li Jingyun. Establishment of nasal mucosal epithelial organoids and assessment of their barrier function[J]. Journal of Capital Medical University, 2026, 47(4): 707-714.

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鼻黏膜上皮类器官的构建及屏障功能分析

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

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