Development of a Serum-free, Three Dimensional Cell Culture Model for Primary Human Articular Chondrocytes to Test Potential Therapeutics

Chondrocytes were isolated by protease digestion and cultured in alginate at 2, 5, 10 x 10 cells in serum-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 5 x 10 cells/culture, as determined by Lowry protein analysis. Type I collagen telopeptide breakdown products were higher in cultures established from the femoral condyles than from the patella, and in OA cultures compared to AKA controls. Type II collagen telopeptides were also detected in both OA and TKA cultures; however, the amounts of these telopeptides were not significantly different in the two types of cultures. 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.