To determine if the exogenous expression of the human telomerase reverse transcriptase (hTERT) protein can extend the in vitro lifespan of chondrocytes from normal and osteoarthritic canine donors, articular chondrocytes were harvested and expanded initially in monolayer culture. Cells were transfected with pCIneo or pCIneo-hTERT and selected using G418. Transfectants were cultured either in monolayer or alginate beads and telomerase activity, replicative lifespan and the tumourogenic potential of the transfected cells were assessed. hTERT expression in canine chondrocytes prolonged the replicative lifespan of these cells but did not permit growth in low serum conditions or promote the formation of foci in anchorage independence assays. In addition, hTERT expression resulted in the down-regulation of MMP-1. This suggests that hTERT may represent a tool for the generation of tissue engineered chondrocytes suitable for autologous re-implantation into the affected areas of osteoarthritic joints.