Bronchial epithelial cell growth supplement

Primary human bronchial epithelial cells (PBEC) were isolated from tumor-free resected bronchus rings obtained from lung cancer patients undergoing a resection surgery at the Leiden University Medical Center (LUMC, Leiden, the Netherlands). Patients from which this lung tissue was derived were enrolled in the biobank via a no-objection system for coded anonymous further use of such tissue (www.coreon.org). Within this framework, individual written informed consent is not needed. Since 01-09-2022, patients are enrolled in the biobank using active informed consent in accordance with local regulations from the LUMC biobank with approval by the institutional medical ethical committee (B20.042/Ab/ab and B20.042/Kb/kb). hPBEC were thawed in a T75 flask coated with 30 ​μg/mL bovine type-1 collagen (PureCol®, Advanced BioMatrix), 10 ​μg/mL fibronectin (Promocell, PromoKine, Bio-connect) and 10 ​μg/mL BSA (ThermoFisher Scientific) in BEpiCM-b basal medium (ScienCell Research Laboratories, Sanbio), supplemented with 100 U/mL penicillin (Lonza), 100 ​μg/mL streptomycin (Lonza) and bronchial epithelial cell growth supplement (ScienCell). When reaching ∼90% confluency, cells were trypsinized in 0.03% (w/v) trypsin (ThermoFisher Scientific), 0.01% (w/v) EDTA (BDH, Poole, UK), 0.1% glucose (BDH) in phosphate buffered saline (PBS) and prepared for seeding.