Deshmukh, A., Arfuso, F., Newsholme, P. and Dharmarajan, A.
Notes: Cancer stem cells (CSCs; 10,000 cells/well in 96-well plates) were washed with PBS and 50µl of 1mM 2-Deoxy-d-Glucose (2DG) added. After 90 minutes, glucose uptake was measured using the Glucose Uptake-Glo™ Assay. CSCs were isolated from cell lines and plated at 10,000 cells/well in 96-well plates. After growing in defined medium supplemented with various amounts of glucose plus glutamine and growth factors for 2 days, cells were treated for 24 hours and 2µl samples removed and added to 98µl of PBS. Extracellular metabolites were assessed using the Glutamine/Glutamate-Glo™ Assay. Isolated CSCs were plated at 10,000 cells/well in 96-well plates, grown for 3 days, treated for 24 hours, medium removed and cells lysed. The lysates were treated to preserve NAD+ and NADH before mixing with the NAD/NADH-Glo™ Detection Reagent and luminescence measured. (4959)