Glucose or lactate assay kit

Lymphoma cell lines were seeded (5 × 10⁵/ml cells) in phenol-free culture media and incubated for 24 hrs. The culture media was collected to measure glucose consumption and lactate production. Glucose and lactate levels were determined using a Glucose or Lactate assay kit (Biovision, San Francisco, CA) according to manufacturer instructions. Glucose consumption was determined by correcting the difference between glucose concentration in cell-free media and the supernatant collected from each cell line culture. Cell numbers were counted with an Automated Cell Counter (Invitrogen Inc, Grand Island, NY) to normalize the glucose and lactic acid concentration according to manufacturer instructions. In addition, differences in ATP production over time between RSCL, RRCL and TRCL were determined using the CellTiter Glo cell viability assay kit from Promega (Madison, WI) following the manufacturer’s instructions. Results were standardized to the number of cell/well and repeated in triplicate.

Viability of cells was determined by resazurin reduction assay in a Chameleon MicroplateReader (Hidex, Turku, Finnland) 31 (link). Extracellular Glucose- and Lactate-amounts were measured after cultivation of identical cell numbers for 72 hours by colorimetric method using the Glucose or Lactate Assay Kit (BioVision, Mountain View, USA) according to the manufacturer's instruction. All levels were normalized to cell numbers.