Glioma Initiating Cells Characterization

Both GICs belong to non-mutated and non-G-CIMP (G-CIMP-) subtypes. Figure 1 displays the optical microscopy images in the medium with and without laminin coated surface. GICs grow as neurospheres in non-laminin coated plates and in adherence in laminin coated surface. Cells placed on 7,5 mg/mL laminin-coated plates (Sigma, St. Louis, MO, USA) were maintained in a complete Neuronal Stem Cell (NSC) medium at 37 °C in a humidified 5% CO₂ and 5% O₂ atmosphere (hypoxia conditions) to simulate brain microenvironment (Heracell 150i incubator). NSC medium was constituted by Dulbecco’s Modified Eagle Medium and Nutrient Mixture F-12,DMEM/F12, (Gibco, Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with N₂ (GIBCO), 4,5% glucose (Sigma, Merck KGaA, Darmstadt, Germany), 1M Hepes (Sigma), 2% BSA (Sigma) basic fibroblast growth factor 20 ng/mL (Gibco), and epidermal growth factor 20 ng/mL (Gibco).
GICs were treated with 200 µM or 2mM of Na[o-COSAN] to study the uptake of the anionic small molecule [o-COSAN]⁻ and its effects on cell function.
Glioma initiating cells (GICs), proneural GIC7 (a kind gift from Dr. Marta María Alonso, Department of Pediatrics, Clínica Universidad de Navarra, University of Navarra, Pamplona, Spain) [74 (link)], and mesenchymal PG88 cells were obtained from human GBM specimens as described previously [46 (link),75 (link)].