Chalcone's Impact on Glioblastoma Clonogenicity

The effect of chalcones’ clonogenic growth potential was determined using sphere-forming assays. Glio3 and Glio38 neurosphere cell lines [6 (link),7 (link)] were dissociated using Accutase (Gemini Bioscience, Sacramento, CA, USA), filtered using 40-micron filter and single cells were seeded at approximately 50 cells per well in a 96-well plate. Cells were treated with 0.1, 0.25 or 0.5 μM of chalcone or 0.01, 0.05 or 0.1 μM of C-dot chalcone conjugate on day 0 and spheres were manually counted under microscopy on day 14. All experiments were done in triplicate. Representative images were obtained using EVOS XL Core light microscope (Thermo Fisher Scientific, Waltham, MA, USA).