Propagation and Culturing of Patient-Derived Glioma Cell Lines

Human patient-derived glioma cell lines GBM6, GBM12, and GBM39 were propagated in mouse flanks before use in culture [46 (link)]. GBM6 and GBM39 spheroids were generated in human neurobasal media (human Neurocult NS-A Basal Medium; Cat 5750; StemCell Technologies, Vancouver, Canada) supplemented with 20 ng/mL human recombinant EGF (Cat 78006; StemCell Technologies), 10 ng/mL human recombinant beta-FGF (Cat 78003; StemCell Technologies), 0.5% StemPro Neural Supplement proliferation media [Cat A1050801; Thermofisher Scientific), and 1% penicillin/streptomycin). Cells were grown in a humidified incubator at 37 °C and 5% CO₂.