The objective of this study was to establish a culture system of primary human articular chondrocytes with which to test novel drugs for the treatment of osteoarthritis (OA). Articular cartilage was obtained from OA subjects undergoing total knee arthroplasty (TKA) or above knee amputation (AKA). Chondrocytes were isolated by protease digestion and cultured in alginate at 2.5, 5, 10 x 106 cells in serum, hormone, and growth factor free medium. Production of collagen degradation products was measured in days 2 and 5 culture medium and day 5 cell associated matrix. The optimal plating density was 2.5 x 106 cells/mL, as determined by end point analysis. Cell yield and collagen breakdown products were higher in cultures established from the tibial plateau and femoral condyle than from the patella. Proteoglycans, collagen, and collagen breakdown products were detected in all OA cultures. However, the production of collagen breakdown products differed significantly between the side of greatest pathology and contralateral region within each subject. These results demonstrate that human articular chondrocytes survive and are metabolically active when cultured in this serum-free, 3D culture system. The femoral condyles and tibial plateau were the optimal sites for tissue harvesting. With this model system we can effectively measure clinical endpoints for testing new drugs that may reduce tissue destruction in OA.
Available at: http://works.bepress.com/marina_dangelo/26/