Establishing and Characterizing Glioma Cell Lines

Twelve human glioma cell lines (U251, U373, U118, U138, A172, U87MG, SW1783, SNB19, LN229, T98G, SF268, and D54) were purchased from ATCC or DSMZ cell collections and were maintained in Dulbecco’s modified Eagle medium (DMEM) containing 10% FBS, 4 mM glutamine, 100 IU/mL penicillin, and 100 μg/mL streptomycin. To minimize the risk of working with misidentified and/or contaminated cell lines, cells were stocked at very low passages. However, periodically, DNA profiling was carried out by using the AmpFLSTRTM identifillerTM PCR amplification kit from Thermo Fisher following the manufacturer’s instructions. Luciferase-transfected U87MG cells were kindly provided by J.E. Heikkila (Abo Akademi University, Turku, Finland). Three GBM patient-derived stem cell lines (BT12M, BT48EF, BT50EF) were kindly provided by J.G. Cairncross and S. Weiss (Arnie Charbonneau Cancer Institute, University of Calgary, Canada) [45 (link),46 (link)], and GSCs-5 and GSCs-7 [47 (link),48 (link)] were from M. Izquierdo (Universidad Autónoma de Madrid, Spain). Luciferase was inserted in the genome of GSCs-5 cells using the pGL4.13 vector (Promega, Milan, Italy) and the jetPEI DNA transfection method (Polyplus, Illkirch, France). Molecular characteristics of the 12 cell lines used in this report are summarized in Supplementary Table S1.