Perfluorooctane sulfonate affects proliferation and differentiation of pluripotent human teratocarcinoma cells

Cell Cultures Derivatives of the NCCIT teratocarcinoma cell line were cultured in Dulbecco's modified eagle medium (DMEM) (Lonza BioWhittaker, Basel, Switzerland), containing 2 mM glutamine and 4.5 g/L glucose. Media was supplemented with 10% fetal bovine serum (FBS) (Gibco-Invitrogen Corporation, Carlsbad, CA, USA). Cell cultures were maintained at 37 ̊C with 5% CO2 in a humidified atmosphere. NCCIT cultures at 60% confluence were harvested by washing with 10 mL of phosphate saline buffer (PBS) in T-75, treated with 1 mL trypsin0.53 mM EDTA solution (Gibco) for 5 min at 37 ̊C, DMEM (10% FBS) was added, cells centrifuged, and then resuspended in DMEM (10% FBS) for sub-culturing, stock freezing or cell proliferation assays. Stocks were obtained from one large NCCIT batch culture, aliquoted in DMEM (20% FBS, 8% dimethyl sulfoxide), frozen gradually and stored in liquid nitrogen for subsequent tests. Cells were confirmed as being free from Mycoplasma infection by the method of Tang et al. (Tang et al. 2000).