Generation of three-dimensional (3D) glioblastoma spheroids
1. U251 MG cell line human (Sigma Aldrich)
2. DMEM High Glucose; Dulbecco's Modified Eagle Medium (Thermo Fisher Scientific)
3. Fetal bovine serum (FBS)
4. 75-cm2 cell culture flask
5. Sterile seropipettes and pipette tips (1000 μl)
9. Non-treated flat-bottom 6-well plate (Sigma Aldrich)
12. Humidified incubator (37 C, 5% CO2)
Detection of apoptosis in glioblastoma spheroid model
2. 12% Bis Tris polyacrylamide gels
3. Electrophoresis machine
4. Trans-Blot® SD Semi-Dry Cell (Bio-rad)
5. Rabbit Anti-Caspase-3 antibody (Cell Signaling)
6. Goat anti-rabbit antibody-HRP (Merck)
7. Anti-β-actin-HRP (Cell Signaling)
8. Immobilon Western Chemiluminescent HRP Substrate
Assessment of drug response in glioblastoma spheroid model
1. DMEM High Glucose; Dulbecco's Modified Eagle Medium (Thermo Fisher Scientific)
2. Fetal bovine serum (FBS)
3. Temozolomide (Sigma Aldrich)
6. Humidified incubator (37 C, 5% CO2)