Characterization of Pediatric Glioblastoma Cell Lines

Human paediatric glioblastoma (GBM) cell lines SF188 and KNS42 have been extensively characterised previously [1 (link)]. KNS42 harbours a histone H3.3 (H3F3A) G34V mutation whilst SF188 is H3 wild-type [4 (link)]. These cell lines were grown as monolayers in DMEM/F12 Ham’s medium (Life Technologies) supplemented with 10% fetal calf serum (GIBCO) and antibiotics at 37 °C and 5% CO₂. Paediatric pHGG cell lines HSJD-DIPG-012 and HSJD-GBM-002 were grown according to the in house protocol from the Montero lab. Cells were cultured as monolayers in flasks pre-treated with laminin (10 μg/ml, BioLamina). Tumour Stem Medium (TSM) base was prepared with 50% Neurobasal-A Medium, 50% D-MEM/F-12, 1% HEPES buffer solution (1 M), 1% sodium pyruvate MEM Life 100MM(CE), 1% MEM non-essential amino acids solution 10 mM (100×), 1% GlutaMAX-I supplement and 1% antibiotic-antimycotic (100×) (LifeTechnologies). 10% FBS was added fresh to TSM base media for preparing a differentiation TSM media. Cells were grown as monolayers in this media at 37 °C with 5% CO₂.