3D Spheroid Culture of Human Nasal Polyp Cells: A Functional Organoid-Like Model for Assessing Epithelial Restoration in Chronic Rhinosinusitis with Nasal Polyps (CRSwNP).

This chapter describes a reproducible protocol for establishing three-dimensional (3D) spheroid cultures from human nasal polyp epithelial cells obtained from patients with chronic rhinosinusitis with nasal polyps (CRSwNP). The model reproduces key features of epithelial dysfunction characteristic of CRSwNP, while also enabling the study of barrier restoration induced by biologic therapies such as Dupilumab. The cultures, although referred to as spheroids, share structural and functional features with airway organoids, including epithelial motility, polarity, and responsiveness to inflammatory cues. Compared to conventional two-dimensional (2D) cell cultures, this system provides a physiologically relevant platform for investigating epithelial pathophysiology and therapeutic modulation in CRSwNP and other airway diseases.