Androgen loss accelerates brain tumour growth via HPA axis activation

Cell lines The syngeneic mouse GBM model SB28 was provided by H. Okada, and GL261 was obtained from the Developmental Therapeutic Program, NCI. CPA was provided by the Castro-Lowenstein Laboratory, and KR158 was provided by L. Deleyrolle. The mouse bladder cancer cell line MB49 was obtained from the Animal Tumour Core at the Cleveland Clinic. The mouse melanoma B16-F10 cells were a gift from T. Stappenbeck. The GBM patient-derived xenograft cell model L1 was obtained from B. Reynolds (originally from the laboratory of A. Vescovi) and GBM23 was obtained from E. Sulman. After thawing, all cell lines were treated with 1:100 MycoRemoval agent (MP Biomedicals) and regularly tested for Mycoplasma spp. (Lonza). GBM cell lines were maintained in complete RPMI 1640 (Media Preparation Core, Cleveland Clinic) supplemented with 10% FBS (Thermo Fisher), 1% penicillin–streptomycin (Media Preparation Core) and GlutaMAX (Gibco). MB49 and B16-F10 cells were cultured in DMEM (Media Preparation Core, Cleveland Clinic) supplemented with 10% FBS, 1% penicillin–streptomycin, GlutaMAX and sodium pyruvate (Thermo Fisher Scientific). GBM patient-derived xenograft cells were cultured in DMEM/F12 medium (ThermoFisher) supplemented with 1% penicillin–streptomycin (ThermoFisher Scientific), 1% B27 without vitamin A (ThermoFisher), 20 ng ml –1 EGF (R&D Systems) and 20 ng ml –1 FGF (R&D Systems). Cells were cultured in humidified incubators at 37 °C and 5% CO 2 and were not allowed to exceed 15 passages. …