Acknowledging the supportive influence of matrix molecules on the chondrocytic phenotype, we combined heparin and chitosan to develop a novel biomaterial, supporting chondrogenesis. Chitosan had been shown as a promising structural material for a number of tissue engineering applications. Similar to heparin, one of the glycosamino-glycans (GAGs) had been known to exert various influences on the biological activities. In this study we evaluated the potential of heparin/chitosan polyelectrolyte complex materials for controlling the proliferation and differentiation of chondrocytes. The heparin/chitosan polyelectrolyte complex was coated over the polyethyleneimine (PEI) layer precoated on the commercial polystyrene dish. When examined by using human chondrocyte cell line (C28/I2), the heparin/chitosan surfaces supported well not only the cell proliferation but also the chondrocytic functions, such as expression of collagen type II. These results indicated that the heparin/chitosan complex could be used to modulate the activity of chondrocytes in vitro.