Culturing Human Glioblastoma Cell Lines

Human glioblastoma cell lines A172, U87, SNB75 and U251 were obtained from the American Type Culture Collection (ATCC). Cells were cultured in DMEM with 4.5 g/L D-glucose (Gibco, Life Technologies) supplemented with 10% foetal calf serum, 100 units/mL penicillin and 100 μg/ml streptomycin in a humidified incubator at 37°C containing 5% CO₂. 2,5-Dimethyl-celecoxib (DMC) was produced as described earlier (Kardosh et al. 2005 (link); Ahlstrom et al. 2007 (link)). Celecoxib (CXB) was purchased from Key Organics. DMC and CXB were both dissolved in DMSO; the control with only solvent (DMSO) showed no toxicity. RhTRAIL wild-type (WT), DR4-specific TRAIL variant 4C7 and DR5-specific TRAIL variant D269H/E195R (amino acids 114–281) were constructed and produced as described earlier (van der Sloot et al. 2006 (link); Reis et al. 2010 (link)).