All patient-derived GSC cell lines and neural progenitor cells (NPCs) were kindly donated by Dr. Krishna P.L. Bhat (The University of Texas, M.D. Anderson Cancer Center, Houston, TX). GSC11, GSC8–11, GSC20, GSC267, GSC28 were established and widely applied in previous studies (6 (link), 29 (link), 30 (link)); their subtypes had already been identified according to the metagene score for PN or MES subtypes based on Philips and Verhaak gene set, respectively (3 (link), 5 (link)). All cell lines were cultured in medium prepared from DMEM/F12 (10565018; Gibco, USA), 2% B-27 no serum supplement (17,504,044; Gibco, USA), 20 ng/mL human recombinant EGF (236-EG; R&D Systems, USA), and 20 ng/mL human recombinant bFGF (233-FB; R&D Systems) using a 37°C, 5% CO2 environment. Accutase solution (A6964; Sigma–Aldrich, USA)-digested tumor spheres were used for passaging. All cell lines used in the experiments were free of mycoplasma contamination. Poly-L-ornithine solution (P4957; Sigma–Aldrich) and laminin (L4544; Sigma–Aldrich) were used to coat the plates to make the cells adhere to the wall for the experiment.
Free full text: Click here