Nsclc h460 cells

Primary normal human LFs were obtained from Lonza (Walkersville, MD) and were cultured in FGM-2 medium containing 2% fetal bovine serum (FBS), 0.1% human basic fibroblast growth factor, 0.1% insulin and 0.1% gentamicin (Lonza). Early-passage cells (p < 6) were seeded (5 × 10⁵ cells/well) in 6-well plate and exposed to lightly sonicated, BSA-dispersed CNTs (Cheap Tubes Inc.), including SWCNT, MWCNT and f-MWCNT at various concentrations for 48 h. Vehicle-only exposed cells were used as a control. Non-small lung cancer cell (NSCLC)-H460 cells were obtained from American Type Culture Collection (ATCC; Manassas, VA) and were cultured in RPMI 1640 medium supplemented with 5% FBS, 2 mM L-glutamine, and 100 units/mL penicillin and 100 μg/mL streptomycin (P/S). Human bronchial epithelial BEAS-2B cells were obtained from ATCC and were chronically exposed to low-dose SWCNT (0.02 μg/cm²) for 6 months to generate BEAS-2B/SWCNT cells as previously described13 (link)14 (link)40 (link). They were cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 5% FBS, 2 mM L-glutamine and P/S. All cells were maintained in a humidified atmosphere of 5% CO₂ at 37 °C.

Primary human small airway epithelial cells (SAECs) immortalized with hTERT were kindly provided by Dr. Hei (Columbia University, NY) [68 (link)]. SAECs were cultured in SABM medium supplemented with Clonetics SAGM SingleQuots (Lonza, Walkersville, MD) which contain 0.4% v/v bovine pituitary extract, 0.1% insulin, 0.1% hydrocortisone, 0.1% retinoic acid, 1% bovine serum albumin, 0.1% transferrin, 0.1% triiodothyronine, 0.1% epinephrine, 0.1% human epidermal growth factor and 0.1% gentamicin. Human bronchial epithelial BEAS-2B cells were obtained from American Type Culture Collection (ATCC; Manassas, VA). They were cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 5% fetal bovine serum (FBS), 2 mM L-glutamine, 100 units/mL penicillin and 100 μg/mL streptomycin. Non-small lung cancer cell (NSCLC)-H460 cells were obtained from American Type Culture Collection (Manassas, VA) and were cultured in RPMI 1640 medium supplemented with 5% FBS, 2 mM L-glutamine, and 100 units/mL penicillin/streptomycin. All cells were maintained in a humidified atmosphere of 5% CO₂ at 37°C.