BEAS-2B所用培养基.doc

BEBM培养基（ATCC）(1个) 1.African Dust Storms Reaching Puerto Rican Coast Stimulate the Secretion of IL-6 and IL-8 and Cause Cytotoxicity to Human Bronchial Epithelial Cells (BEAS-2B) BEAS-2B cells were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA; Cat No CRL-9609). Cells were cultured according to ATCC protocols, maintained in keratinocyte basal medium (KBM-2, Lonza, Walkersville, MD, USA Cat No CC 3103) and supplemented with keratinocyte growth medium (KGM-2 SingleQuots; Lonza, Walkersville, MD, USA; Cat No CC4152). Cells were used at passages 44 - 59 and maintained at 37°C in a humidified atmosphere of 5% CO2 DMEM（11） 1.Diatom-Derived Polyunsaturated Aldehydes Activate Cell Death in Human Cancer Cell Lines but Not Normal Cells In an independent experiment, A549 cells (2×103 cells well-1) were seeded in a 96-well plate and kept overnight for attachment. The next day the medium was replaced with fresh medium with three concentrations (2, 5 and 10 μM) for each of three PUAs (DD, OD, and HD, Sigma-Aldrich Inc., Milano, Italy) tested and with caspase-3 Inhibitor (C30H41FN4O12, sc-3075, Santa Cruz) at 9.7 μM; cells were allowed to grow for 24, 48 and 72 h. After aldehyde treatment, viable cells were evaluated as described below. The BEAS-2B (ATCC CRL-9609) lung/brunch normal epithelial cell line was maintained in DMEM (Dulbeccos modified Eagles medium) supplemented with 50% fetal bovine serum (FBS), 100 units ml?1 penicillin and 100 μg ml?1 streptomycin. Cells were incubated in a 5% CO2 humidified chamber at 37°C for growth. BEAS-2B (2×103 cells well?1) was seeded in a 96-well plate and kept overnight for attachment. The next day the medium was replaced with fresh medium with three concentrations (2, 5 and 10 μM) for each of three PUAs (DD, OD, and HD, Sigma-Aldrich Inc., Milano, Italy) tested; cells were allowed to grow for 24, 48 and 72 h. After incubation, the supernatant was removed and adherent cells were examined for viability. 2.Gene Ex