Department of Surgery-Otolaryngology, Head and Neck Surgery, Central Adelaide Local Health Network (Basil Hetzel Institute), The Queen Elizabeth Hospital and The University of Adelaide, Adelaide, Australia.
College of Medicine and Public Health, Flinders University, GPO Box 2100, Adelaide, South Australia 5001, Australia.
Biol Open. 2022 Aug 15;11(8). doi: 10.1242/bio.059267. Epub 2022 Aug 16.
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.
在三维培养中生长的患者来源的类器官为表型高通量筛选和药物反应研究提供了极好的平台。类器官技术已被应用于研究干细胞生物学和各种人类病理学。本研究调查了源自慢性鼻-鼻窦炎（CRS）患者的原代人鼻腔上皮细胞（HNECs）的类器官的特征和细胞形态。对鼻腔类器官进行培养，最长可达 20 天，并通过免疫荧光、RT-qPCR、western blot 和 FACS 分析测量形态学、细胞组成和功能参数。结果表明，鼻腔类器官表达干细胞标志物富含亮氨酸重复的 G 蛋白偶联受体 5（LGR5），以及顶端连接基因、杯状细胞和纤毛细胞的标志物。此外，我们能够在冷冻和解冻后很好地重新生长和扩大鼻腔类器官。本研究为开发人鼻腔类器官提供了一种有效且可行的方法，适用于表型高通量筛选和药物反应研究。
