Objective To study the separation and cultivation of human osteoarthritis chondrocytes.Methods Cartilage slices were harvested from 10 osteoarthritis patients during arthroscopic surgery.The cartilage slices were digested with 0.2% collagenase Ⅱ and 0.25% trypsin,and cultured in monolayer cell in vitro.During the cultivation,the chondrocytes' shape,growth and proliferation were observed.Results Chondrocyte could be effectively isolated by digesting with collagenase Ⅱ and trypsin.The osteoarthritis chondrocytes proliferated slowly and showed weak heterochromia to toluidine blue and immunohistochemical stain of collagen type Ⅱ.Conclusions It is an effective method to digest cartilage by collagenase Ⅱ and trypsin.The cultured chondrocyte in vitro has biological characteristics of human osteoarthritis degenerated cartilage and this method could be useful for the diagnosis and treatment of osteoarthritis.