Cultured human fibroblasts isolated from embryonic muscle, skin and peripheral nerve tissues were found to accumulate [H-3]L-glutamate by a Na+-dependent uptake process strongly inhibited by several glutamate/aspartate analogues including D- and L-aspartate, D- and L-thleo-3-hydroxyaspartate and L-trans-pyrrolidine-2,4-dicarboxylate but not D-glutamate. It was also reduced by elevated concentrations of K+, Rb+ and Cs+. The values of K-m's were 5-20 mu M, well within the 'high affinity' region. Variations in the capacity (V-max) of [H-3]L-glutamate uptake did not correlate with the origin (muscle, skin or nerve tissue) of the fibroblasts. The uptake characteristics suggest that it is mediated by a transport system similar to that commonly observed only in brain tissue.