Dmem f 12 glutamax media

LN18, LN229, T98G (T98), U87-MG (U87) and U251 cell lines were purchased from the ATCC, USA, and cultured in DMEM medium (Dulbecco’s modified Eagle medium, Thermo Fisher Scientific, USA). Primary glioblastoma cell lines (WG4 and WG9) were derived as described [43 (link)] and cultured in DMEM/F12 GlutaMAX media (Thermo Fisher Scientific, USA). Normal human astrocytes (NHA, Lonza) were cultured in medium containing ABM^TM Basal Medium (CC-3187) and AGM^TM SingleQuots^TM Supplements (CC-4123) from Lonza.. All culture media were supplemented with 10% FBS (Gibco, USA), antibiotics (100 U/mL penicillin, 100 µg/mL streptomycin) and cultured in a humidified atmosphere of CO₂/air (5%/95%) at 37 °C. Cells were treated with Temozolomide (Sigma, USA) alone or with combination with: Olaparib, 3-aminobenzamide, Rucaparib, Tioguanine or Etoposide. Aforementioned compounds were dissolved in DMSO. Irradiation of UV-C light was used with 30 J/m² dose. For reagent specifications and catalogue numbers see the Supplementary Table S1.

Human iPSCs were differentiated into CD31 + endothelial cells^{40 (link)}. For endothelial differentiation, iPSCs were dissociated into single cells and seeded onto Matrigel-coated plates at a density of 1 × 10⁵ cells/cm² in TeSR-E8 media supplemented with 10 μM Y − 27632. After 24 h, referred to as day 0, media was replaced with DMEM/F12 GlutaMAX media (Thermo Fisher Scientific) containing N-2 supplement (Thermo Fisher Scientific), B27 supplement, 8 mM CHIR99021 and 25 ng/mL BMP4 (STEMCELL Technologies) for 3 days. Media was then replaced with StemPro-34 SFM complete media (Thermo Fisher Scientific) supplemented with 200 ng/mL VEGF-165 (Peprotech) and 2 µM forskolin (Sigma-Aldrich) for 3 days. At day 6, hiPSC-derived endothelial cells were dissociated into single cells, resuspended in fluorescence-activated cell sorting (FACS) buffer (0.5% (w/v) BSA, 2 mM EDTA in DPBS) and incubated with conjugated FITC Mouse Anti-Human CD31 (1:10 dilution, #555445, clone WM59, BD Pharmingen, lot#8212882) for 30 mins. Stained cells were resuspended in FACS buffer, passed through a 40 µm mesh, and sorted using a BD FACS Aria III (Supplementary Fig. 34). An unstained cell sample was used as a negative control for gating. CD31 positive cells expanded on human fibronectin (Merck) coated plates and cultured in EGM2-MV media (Lonza) supplemented with 50 ng/mL VEGF-165.

Mouse embryonic fibroblasts obtained from (Cyagen, Suzhou, China). Cells were cultured in DMEM/F12 + GlutaMAX media (Gibco, Waltham, MA, United States) plus 10% fetal bovine serum (Excell, St. Louis, MO, United States), 100 U/mL streptomycin/penicillin (Gibco, Waltham, MA, United States), 1% 100 × NEAA (Sigma, St. Louis, MO, United States) at 37°C in a 5% CO₂ cell incubator.