After a careful dissection of trabecular tissue from recent postmortem specimens, human trabecular cells were established in tissue culture with 250 ng/ml fibroblast growth factor (FGF), Dulbecco's modified Eagle's medium (DME), and 10% human serum. These conditions have allowed propagation of human trabecular cells for a number of passages at high density without apparent cellular degeneration. FGF increased the rate of cell division and the plating efficiency for trabecular cells but was not needed after cells had achieved confluency. Human trabecular cells had a pattern of growth which differed from human corneal keratocytes and human corneal endothelial cells compared at a similar passage. Propagation of human trabecular cells in vitro may provide a valuable source of experimental material to study the functional aspects of these cells which line the trabecular meshwork.