Harvesting and Culturing Human Nasal Epithelial Cells

HNECs were harvested from the inferior turbinate using a nasal brush and cultured as described (Ramezanpour et al., 2016 (link), 2017 (link)). Briefly, extracted HNECs were suspended in Bronchial Epithelial Growth Media (BEGM, CC-3170, Lonza, Walkersville, MD, USA), supplemented with 2% Ultroser G (Pall Corporation, Port Washington, NY, USA). The cell suspension was depleted of monocytes using anti-CD68 (Dako, Glostrup, Denmark) coated culture dishes, and HNECs expanded in routine cell culture conditions of 37°C humidified air with 5% CO₂ in collagen coated flasks (Thermo Scientific, Walthman, MA, USA). HNECs were tested at passage two and confirmed to be of epithelial lineage via reactivity to pan-Cytokeratin and CD45 antibodies (both from Abcam, Cambridge, MA, USA), and a Diff-Quick staining method used in the assessment of cell morphology by professional cytologists (IMVS, The Queen Elizabeth Hospital, Woodville, Australia).