D247mg

Human GIC (S-24, T-269, T-325, ZH-161, and ZH-305) were established from freshly resected tumors [62 (link)]. The human long-term glioma cell lines LN-18, LN-428, D247MG, LN-319, A172, LN-308, and LN-229 [63 (link)] were kindly provided by N. de Tribolet (Lausanne, Switzerland) and T98G cells were obtained from the American Type Culture Collection (ATCC) (Rockville, MD). GIC were cultured as neurospheres in Neurobasal medium (NB) supplemented with 2 mM l-glutamine, 20 μg/ml B-27 supplement (Gibco, Waltham, MA), 20 ng/ml fibroblast growth factor (FGF) 2, and 20 ng/ml epidermal growth factor (EGF) (Peprotech, Rocky Hill, PA). Long-term glioma cell lines were grown as adherent monolayers in Dulbecco´s modified Eagle´s medium (DMEM) supplemented with 10% fetal calf serum (FCS) and 2 mM l-glutamine (Gibco). Cells were regularly tested for mycoplasma contamination by means of MycoAlert^TM PLUS Mycoplasma Detection (Lonza, Basel, Switzerland).

Eight human malignant glioma cell lines (LN-18, LN-229, LN-308, LN-428, U87MG, U251MG, U373MG, and D247MG) were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA). All cell lines were cultured in Dulbecco's modified essential medium (DMEM, Invitrogen Corp., Grand Island, NY, USA) and supplemented with 10% heat-inactivated fetal bovine serum (Invitrogen Corp., Grand Island, NY, USA), 2 mM glutamine, and gentamicin at 37°C in a humidified incubator with 5% CO₂. Experiments were performed in triplicate.