What question did this study set out to answer?

The aim is to evaluate how different culturing methods impact the physiology of human nasal epithelial cells.

March 25, 2026Open Access

Nasal organoids as optimal models for studying structure and function of primary nasal epithelial cell cultures

MRMahnaz Ramezanpour GBGeorge BourasFlinders University SFSholeh FeiziHealth Net

Key Points

The aim is to evaluate how different culturing methods impact the physiology of human nasal epithelial cells.
Collected HNECs from nasal brushings of control and CRSwNP patients.
Cultured cells under four conditions: monolayers, air-liquid interface, Dome organoids, and ALI organoids.
Applied mass spectrometry for proteomic profiling to analyze protein expression.
Conducted gene set enrichment analyses comparing healthy and CRSwNP samples.
Organoid cultures resembled nasal brushings more closely than other methods.
ALI organoids had increased cilia-related protein expression and superior cilia beat frequency.
Dome organoids displayed varied orientation of differentiated cells compared to ALI organoids.

Abstract

Primary cell cultures serve as representative model systems for studying the normal physiology and biochemistry of cells. However, various methods exist for culturing these cells, and it remains unclear how these methods impact cell physiology. Participants included both non-CRS control individuals and patients diagnosed with chronic rhinosinusitis with nasal polyps (CRSwNP). Human nasal epithelial cells (HNECs) were collected from nasal brushings and cultured under four different conditions: monolayers, air-liquid interface (ALI) cultures, Dome organoids, and ALI organoids, resulting in a total of 40 samples, including nasal brushings. We utilized the latest advances in mass spectrometry (MS) technology to gain new insights into how these different culture methods affect the protein expression of HNECs. Gene set enrichment analyses comparing nasal brushings from healthy and chronic rhinosinusitis with nasal polyps (CRSwNP) patients to HNECs cultured in various conditions showed that organoid cultures closely resemble nasal brushings. Ultrastructural analysis revealed opposing orientations of differentiated cells in Dome and ALI organoids, with ALI organoids showing increased expression of cilia-related proteins and cilia positioned on the external surface of the organoids. Both ALI and Dome organoids contained ciliated cells; however, cilia beat frequency measurements were more consistent and uniform in ALI organoids compared to Dome organoids. This research could contribute to future studies aiming to improve our understanding of the pathophysiology and treatment of CRS. • Proteomic profiling compared four HNEC culture models with nasal brushings. • Organoid cultures show fewer perturbed pathways and closer in vivo similarity. • ALI organoids exhibit apical-out orientation with enhanced ciliogenesis. • ALI organoids display more consistent cilia beat frequency than Dome cultures. • ALI organoids enable improved access to the apical surface for functional studies.

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