To examine whether the life span of fibroblast-like synoviocytes (FLSs) can be extended and to establish FLS cell lines that preserve the characteristics of primary FLSs, we introduced human catalytic subunit of telomerase (hTERT) gene into human osteoarthritic (OA) FLSs. Two hTERT-transduced clonal cell lines were established and one line, hTERT-OA FLS 13A, was characterized. The hTERT-OA FLS 13A cells have a morphology similar to that of the parental untransduced cells and a population-doubling time similar to that of the parental cells of early passages. While the parental untransduced OA FLSs reached senescence after 100 days in culture, the hTERT-OA FLS 13A cells continued to grow at a population-doubling rate of once in about every 2-3 days. The hTERT-OA 13A cells have so far grown in culture beyond 450 days and maintained the same growth rate. Furthermore, the hTERT-OA FLS 13A cells preserved their sensitivity and response to the treatment with basic calcium phosphate crystals and interleukin-1beta. In conclusion, exogenous expression of telomerase represents a way to extend the life span of human FLSs and telomerase-transduced FLS cells offer a promising tool for gene regulation, cell-based assay, cell transplantation-based gene therapy, and tissue engineering research and development.