| GSM8397410 channel 1 treatment |
Patient-derived cultures OPBG-GBM-001 (H3.3 G34R), HSJD-GBM-002 (H3.3 G34R) and CHOP-GBM-001 (7316-158; H3.3 G34R) were grown in stem cell media consisting of Dulbecco’s Modified Eagles Medium: Nutrient Mixture F12 (DMEM/F12), Neurobasal-A Medium, 10 mM HEPES Buffer Solution, 1 mM sodium pyruvate solution, 1% (v/v) non-essential amino acids solution (100x) and 1% (v/v) Glutamax-I supplement (100x). The media was supplemented with 2% (v/v) B-27 supplement minus vitamin A (50x), 20 ng/ml Human-EGF, 20 ng/ml Human-FGF-basic-154, 20 ng/ml Human-PDGF-AA, 20 ng/ml Human-PDGF-BB (all Shenandoah Biotech, Warwick, PA, USA) and 2 μg/ml Heparin Solution (Stem Cell Technologies, Cambridge, UK). Cells were cultured on Cultrex® 3D Culture Matrix Laminin (Bio-Techne Ltd) except for CHOP-GBM-001 cells, which were cultured on Geltrex (Invitrogen). KNS42 cells (H3.3 G34V) were obtained from the Japanese Collection of Research Bioresources Cell Bank (#IFO50356) and grown in DMEM/F12 with 10% (v/v) FBS. HEK293FT (RRID:CVCL_6911) cells were purchased from Invitrogen and grown in DMEM containing 10% (v/v) FBS. BT690 (H3.3G34R) were grown in NSA Working Media (Stem Cell Technologies, #5751), supplemented with 20 ng/ml Human-EGF and 20 ng/ml Human-FGF, and 2 µg/ml Heparin Solution. Cells were incubated at 37℃ in a humidified atmosphere at 5% CO2. |
1 |