HyClone Dulbecco's Modified Eagle Medium (DMEM) with high glucose: Liquid

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Cells and cell culture conditions: The human GBM cell lines U87MG (RRID, CVCL_0022, glioblastoma of unknown origin), LN229 (RRID, CVCL_0393), LN18 (RRID, CVCL_0392), T98G (RRID, CVCL_0556), A172 (RRID, CVCL_0131) and A1207 (RRID, CVCL_8481) were purchased from the American Type Culture Collection. Normal human astrocytes (NHAs) were purchased from ScienCell Research Laboratories, Inc. Human GBM cells and NHAs were cultured in high-glucose DMEM supplemented with 10% FBS, 1% penicillin/streptomycin and 2 mM L-glutamine (all from HyClone; GE Healthcare Life Sciences) and placed at 37°C in a humidified incubator containing 5% CO2 and 95% humidity. PN-GSCs [528NS, GSC11 (RRID, CVCL_DR55) and GSC23 (RRID, CVCL_DR59)], MES-GSCs (GSC20) and non-defined GSCs (GSC28) were kindly provided by Dr Ichiro Nakano of Ohio State University (528NS) and Dr Erik Sulman of The University of Texas M.D. Anderson Cancer Center (GSC11, GSC23, GSC20 and GSC28). GSCs were cultured in DMEM/F12 (HyClone; GE Healthcare Life Sciences) supplemented with 0.04% B27 (Invitrogen; Thermo Fisher Scientific, Inc.), 20 ng/ml epidermal growth factor (EGF; R&D Systems, Inc.), 20 ng/ml basic fibroblast growth factor (bFGF; R&D Systems, Inc.), 1% penicillin/streptomycin (HyClone; GE Healthcare Life Sciences) and 2 mM L-glutamine (HyClone; GE Healthcare Life Sciences) and placed at 37°C in a humidified incubator containing 5% CO2 and 95% humidity. Mycoplasma testing was performed using reverse transcription-quantitative PCR (RT-qPCR). All human cell lines were authenticated using short tandem repeat profiling.

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