Mycoaler kit

Human lung cancer cell lines with wild-type EGFR (H358, H23, A549, H441, H69, Calu-6, and H460) (ATCC) and with EGFR mutations (H1650, H1975, HCC827, PC9, PC9-ER, and H3255) (provided by Dr. Susumu Kobayashi) were cultured in DMEM (high glucose) (Gibco) with 10% fetal bovine serum (FBS), 2 mM L-glutamine and 1% penicillin–streptomycin. Primary lung fibroblast CCD-13Lu (ATCC) and rat alveolar macrophage NR8383 (ATCC) were cultured in DMEM (high glucose) (Gibco) with 10% FBS, 2 mM l-glutamine and 1% penicillin–streptomycin. Human lung microvascular endothelial cell HULEC-5A (ATCC) was cultured in MCDB131 (Gibco) supplemented with 10 ng/ml epidermal growth factor (EGF)(Gibco), 1 μg/ml hydrocortisone (Stemcell), 10 mM l-glutamine and 10% FBS. Immortalised tracheobronchial epithelial (AALE) cells were derived as previously described and maintained in SAGM media (Lonza) [60 (link)]. Cell line identities were confirmed by STR fingerprinting and all were found negative for mycoplasma using the MycoAler Kit (Lonza).

Each cell line was maintained in a 5% CO₂ atmosphere at 37 °C. Human lung EGFR-mutant cell lines H1650, H1975, HCC827, PC9, and H3255 (provided by Dr. Susumu Kobayashi) were cultured in DMEM (high glucose) (GIBCO) with 10% FBS, 2 mM l-glutamine and 1% penicillin–streptomycin. Immortalized tracheobronchial epithelial AALE cells (provided by William C. Hahn) were derived as previously described [100 (link)] and maintained in SAGM media (Lonza). Cell lines were negative for mycoplasma using the MycoAler Kit (Lonza).