Unscheduled DNA synthesis (UDS) was demonstrated both in fresh slices of rabbit articular cartilage and in resting chondrocytes dissociated from it. Uptake of [3H]thymidine by chondrocytes treated with 10 mM hydroxyurea and irradiated with 254 nm ultraviolet light (20 J/m2) was measured by autoradiography. The UDS in the resting chondrocytes was minuscule compared with that of chondrocytes growing in monolayer culture. It was greater in resting cells from 3- than in 18-month old rabbits but persisted in animals up to 52 months of age. In both resting and cultured chondrocytes, UDS was greatly inhibited by aphidicolin (5 micrograms/ml). The finding of age-related decrease in UDS of resting chondrocytes, contrasted with previously demonstrated lack of it in dividing ones, raises questions about in vitro artefacts in studies of cellular senescence.