Characterization of human nasal organoids from chronic rhinosinusitis patients

慢性鼻窦炎患者鼻腔类器官的表征

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作者:Mahnaz Ramezanpour, Harrison Bolt, Karen Hon, Gohar Shaghayegh, Hadi Rastin, Kevin Aaron Fenix, James Psaltis Alkis, Peter-John Wormald, Sarah Vreugde

Abstract

Patient-derived organoids grown in three-dimensional cultures provide an excellent platform for phenotypic high-throughput screening and drug-response research. Organoid technology has been applied to study stem cell biology and various human pathologies. This study investigates the characteristics and cellular morphology of organoids derived from primary human nasal epithelial cells (HNECs) of chronic rhinosinusitis (CRS) patients. Nasal organoids were cultured up to 20 days and morphological, cell composition and functional parameters were measured by immunofluorescence, RT-qPCR, western blot and FACS analysis. The results showed that nasal organoids expressed the stem cell marker leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5), and markers for apical junction genes, goblet cells and ciliated cells. Moreover, we were able to regrow and expand the nasal organoids well after freezing and thawing. This study provides an effective and feasible method for development of human nasal organoids, suitable for the phenotypic high-throughput screening and drug response research.

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