Immortalization of Fibroblasts and ADC-TAFs

Control fibroblasts and ADC-TAFs from randomly selected patients (#5, #13, #37) were immortalised with hTERT as reported [9 (link)]. Unless otherwise indicated, all fibroblast experiments were performed in culture medium containing serum-free high-glucose (4.5 g/l) DMEM supplemented with 1% insulin–transferrin–selenium (ITS) and antibiotics as described [10 (link)]. In some experiments, fibroblasts were stimulated with 2.5 ng/ml recombinant human TGF-β1 (Miltenyi Biotec) at different time points as indicated, which is similar to the average TGF-β1 concentration reported in the bronchoalveolar lavage fluid of lung cancer patients [21 (link)].