Establishment of human nasolacrimal duct epithelial organoid model.

INTRODUCTION: The nasolacrimal duct (NLD) holds significant clinical importance in diseases represented by primary acquired NLD obstruction (PANDO). This study aims to culture human NLD epithelial cells using organoid culture systems, with the goal of establishing cell models that recapitulate the in vivo characteristics of the NLD in an in vitro setting. METHODS: Human NLD epithelial specimens were surgically obtained from PANDO patients during endoscopic dacryocystorhinostomy. Human NLD epithelial cells were isolated via enzymatic digestion and expanded using Expansion Medium. Air-liquid interface culture was performed to induce differentiation, evidenced by active ciliary beating within 28 days. Both apical-in and apical-out NLD organoids were successfully generated using Matrigel-based 3D culture systems and AggreWell™ 400 24-well plate. RESULTS: Histological analysis confirmed that the human NLD epithelium comprised a stratified columnar architecture with ciliated, goblet, and basal cells. Using an air-liquid interface system, primary epithelial cells were successfully differentiated into a polarized layer exhibiting native tissue characteristics. Furthermore, both apical-in and apical-out organoids were established, recapitulating key structural and cellular features of the native NLD epithelium. CONCLUSION: This NLD epithelial organoid system provides a versatile platform for studying NLD physiology, pathology, drug screening, and naso-ocular interactions, with potential for further expansion into broader research applications.