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The metabolism of [1-13C] glucose was followed in C6 rat glioma cells immobilized on a gel thread and in perchloric extracts of the same cells in culture. The results showed that the main metabolite of [1-13C] glucose is [3-13C] lactate. The effects of hypoxia were followed in the perchloric acid extracts of C6 cells. In normoxic conditions, the main metabolites produced by the cells were [3-'3C] lactate, [3-13C] alanine, [2-13C], [3-13C] and [4-13C] glutamate. Lactate newly synthesized from glucose appeared to be exported in the perfusion medium when living cells were immobilized in gel threads made of extracellular matrix. After 5 h of hypoxia, the lactate labelling measured in PCA cell extracts was increased that of glutamate decreased and the appearance of a spectral line at 66.01 ppm, identified as [1-13C] glycerol-3-phosphate, was observed. The data suggest that the synthesis of glycerol-3-phosphate in these cells might represent a sign of hypoxia.