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)

Notes: The authors of this paper describe bioluminescent glucose, lactate, glutamine and glutamate assays suitable for high-throughput analysis of two major metabolic pathways in cancer cells: glycolysis and glutaminolysis. (4769)

Notes: These authors report that glutamine promotes proliferation of cancer cells independently of its use as a metabolic fuel or as a precursor of glutathione, and that glutamine also activates transcription factors hypoxia-inducible factor-1, mammalian target of rapamycin and c-Myc, but these factors do not mediate the effects of glutamine on cancer cell proliferation. During the study, total RNA was isolated from cells using the Maxwell® RSC instrument,  ATP levels were measured using the CellTiter-Glo® Assay and intracellular glutamine and glutamate were measured using the Glutamine/Glutamate-Glo™ Assay. HIF-1 and PHD activities were measured using the Dual-Luciferase® Reporter Assay.