Periosteum-derived progenitor cells(PDPCs) have a capacity to differentiate into a number of connective tissue lineage. They are an attractive source of chondrocyte-progenitor cells. When these cells are cultured in a threedimensional format in the presence of transforming growth factor-b3(TGF-b3), they showed characteristic morphological changes with the deposition of cartilaginous extracellular matrix(ECM). In this study, effects of various cartilage-specific ECMs or feeder cells on the chondrogenesis of PDPCs were investigated. Application of this system resulted in a more functional format and enhanced the cell viability than the chondrogenesis without cartilage-specific ECMs or feeder cells. Differentiation was found to be dependent on immunophenotypes and was interrelated with passage number. FACS analysis on CD29, CD44, CD105 by serial passage numbers of PDPCs provided an optimal seeding time for chondrogenesis. Addition of chondroitin 6-sulfate, known as cartilage-specific ECM, in induction media enhanced chondrogenesis, as indicated by the synthesis of glycosaminoglycan(GAG). When chondrocytes were used as feeder cells, the cell viability was increased during chondrogenesis.