Culturing Diverse Respiratory Cell Lines

The stable cell line MN8CampLuc-HT-29 was cultured as described previously^{14 (link)} except addition of antibiotics and 10% FBS instead of FCS was used. The parental HT-29 cell line (ATCC: HTB-38, batch 70019050) was cultured in McCoy’s 5A (modified) medium (Life Tech., Cat. No. 26600-023) supplemented with 10% FBS (Life Tech., Cat. No. 10270-106), otherwise indicated. Human bronchial/tracheal epithelial primary cells (HBEpC) from two different donors labeled #2644 and #1865 were provided from Cell Applications (Cat. No. 502-05a). BCi-NS1.1^{35 (link)}, VA10^{36 (link)} and HBEpC cells were cultured in Bronchial/Tracheal Epithelial cell growth medium (BEGM) supplemented with retinoic acid (Cell Applications, 511A-500 and 511-RA). HBEpC were sub-cultured using subculture reagent kit (Cell Applications, 090K) and following instructions provided by the vendor. Experiments with SBI-115 antagonist were performed by culturing BCi cells in BEGM BulletKit from Lonza (Cat. No. CC-3170), as BEGM from Cell Applications was temporarily not available from the vendor. Cells were cultured at 37 °C and 5% CO₂.