Stable F508del CFTR cell lines for research

A CFBE41o- cell line stably expressing F508del CFTR harboring an HRP-tag in the fourth extracellular loop was obtained from Professor Gergely Lukacs (Department of Physiology, McGill University, Montreal, QC, Canada) (Veit et al., 2012 (link)). Cells were grown in Eagle’s minimal essential medium (MEM) (Life Technologies) supplemented with 10% FBS, 1% L-glutamine (Life Technologies), 10 mM HEPES (Life Technologies), 200 μg/ml geneticin (Life Technologies) and 3 μg/ml puromycin (Sigma) in culture flasks coated with 0.01% bovine serum albumin (BSA) (Sigma), 30 μg/ml Purecol (Nutacon) and 0.001% human fibronectin (Sigma). HEK293 cells were cultured in uncoated flasks using Dulbecco’s Modified Eagle Medium (DMEM) (Life Technologies) supplemented with 10% FBS and 1% penicillin/streptomycin. The U2OS EA-MEM F508delCFTR cell line expresses the larger beta-galactosidase EFC fragment localized in the plasma MEMbrane (EA: enzyme acceptor) and CFTR with the EA-MEM fusion protein, and was obtained from DiscoverX. These cells were cultured in a medium developed by DiscoverX (assay complete medium, 92-0018GK3). CHO cells were cultured in DMEM containing 10% FBS.