Hela h1

The epithelial cell line 16HBE14o-, derived from human bronchial epithelial cells (Prof. D.C. Gruenert, University of California, San Francisco, CA, USA), was used as a surrogate for the respiratory epithelium. The cells show properties of differentiated airway epithelial cells including formation of tight junctions, apical microvilli and cilia and form polarized monolayers when grown on Transwell filters [44 (link)]. The 16HBE14o- cells were cultured at 37 °C in a humidified atmosphere containing 5% CO₂ in Minimum Essential Medium (MEM; Gibco, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (FBS; HyClone, GE Healthcare, Buckinghamshire, UK), 100 U/mL penicillin and 100 µg/mL streptomycin (Gibco, Thermo Fisher Scientific). Cells were cultured in flasks coated with LHC basal medium (Gibco, Thermo Fisher Scientific) supplemented with 100 µg/mL BSA, 30 µg/mL collagen and 10 µg/mL fibronectin (BD Biosciences, San Jose, CA, USA).
The cervical epithelial cell line H1HeLa (American Type Culture Collection, ATCC, Manassas, VA, USA) was grown at 37 °C in a humidified atmosphere containing 5% CO₂ in MEM supplemented with 10% FBS (Gibco, Thermo Fisher Scientific) and antibiotics (100 U/mL penicillin, 100 µg/mL streptomycin and 1.2 µg/mL gentamycin (Gibco, Thermo Fisher Scientific).

Human cervix carcinoma cells expressing ICAM-1 (HeLa-H1, CRL 1958; ATCC, Manassas, VA) were maintained in suspension culture or as attached cells. For suspension cultures, cells were maintained in complete DMEM (8% FBS, 1 mM EDTA, 1% pluronic F68, and 1% penicillin/streptomycin) within Erlenmeyer flasks shaken at 100 rpm at 37 °C in a 5% CO₂ atmosphere. Cultures were seeded at a concentration of 5 × 10⁵ cells/mL and subdivided every 3–4 days. Monolayer cell cultures were maintained in DMEM without phenol red supplemented with 10% FBS, 30 mM MgCl₂, and 1% penicillin/streptomycin. Cells were subdivided before confluence.