The viability of cartilage explants preserved in vitro decreases with time, which limits its use for transplantation. The effect of mechanical stimulation to cartilage explants in vitro is unknown. In this study, we observed the effects of mechanical stimulation on chondrocyte viability and the mechanical properties of cartilage explants preserved in vitro using a rolling-sliding loading device designed by us, and the optimal stimulation protocol was established. A cylindrical osteochondral mass drilled on the femoral condyle of a healthy pig was divided into two groups (loading group and control group), and changes in the chondrocyte survival rate, matrix composition and cartilage biomechanical properties was observed at different time points. Additionally, the mRNA expression of the apoptosis-related proteins caspase-3/Bax/Bcl-2, the cytoskeletal proteins actin/vimentin, and the matrix-related protein MMP13 were detected. The loading group exhibited delayed collagen and aggrecan degeneration and improved chondrocyte viability for three days. Protein and mRNA detection showed that apoptotic factors such as caspase-3 and Bax decreased rapidly in cartilage tissue after loading. The cytoskeletal proteins actin and vimentin showed no significant changes in mRNA expression in the control group, but was significantly higher in the loading group. MMP-13 mRNA expression was significantly higher in both the control group and loading group. Overall, this study suggests that suitable mechanical stimulation decreases the loss of chondrocyte viability and the mechanical properties of cartilage explants in vitro and improves cartilage preservation.