Objective To establish a 3D culture model of chondrocytes within agarose gel and evaluate the mechanical properties of the chondrocyte-agarose disks. Methods Chondrocytes were mixed with agarose, serum and medium to form engineered cartilaginous constructs. At the 7th day, 14th day and 21st day, the constructs were removed respectively from the culture plates, and stained with Safranin-O for sulfated glycosaminoglycan or Toluidine Blue for proteoglycans or labeled using antibodies against collagen types II. The mechanical properties of ultimate stress, strain, Young’s modulus and tangent modulus were assessed using Instron5544 machine at the same time. Result Agarose gel was found to provide a good microenvironment for formation of extracellular matrix. In addition, it could construct mature and mechanical properties depended on the culture time. At the 21st day, experimental studies have shown that ultimate stress, Young’s modulus, tangent modulus(0.0226±0.006, 0.608±0.061, 0.096±0.004) exhibited a higher value compared with that of 7th day (0.0204±0.004, 0.558±0.036, 0.029±0.002) and14th day (0.0213±0.008, 0.586±0.095, 0. 049±0.005), but there is no significantly difference in ultimate strain (P>0.05). Conclusion The mechanical properties of chondrocytes/agrose disks depend directly upon the extracellular matrix surrounding the cells.