As an ideal antioxidant and anti-apoptotic substance, hydrogen (H2) has protective effects on many isolated organs, such as the heart, lung and kidney. In this study, we explore whether H2 improves the preservation effect of osteochondral allograft by adding it to Dulbecco’s Modified Eagle Medium (DMEM) solution during the tissue culture stage. The osteochondral allograft apparatus was used to harvest 60 pieces of cylindrical allografts (l = 10 mm, d = 6 mm) cartilage in the lateral loading area of the femoral condyle from the pig knee joint in the aseptic condition, and the grafts were randomly divided into 4 groups: Control group (DMEM solution without hydrogen); H-1 group (DMEM solution with hydrogen concentration of 0.2 mmol/L); H-2 group (DMEM solution with hydrogen concentration of 0.4 mmol/L); and H-3 group (DMEM solution with hydrogen concentration of 0.8 mmol/L). The chondrocyte viability, histological changes (hematoxylin and eosin staining, Safranine O staining, and collagen type II immunohistochemistry staining) and biomechanical properties (Young’s modulus) of the osteochondral allograft were investigated after 28 days’ storage. The chondrocyte viability and proteoglycan and collagen type II contents in the H-3 and H-2 groups were higher than that in the Control and H-1 groups, and the H-3 group had the highest values. However, significant differences were not observed between the four groups based on Young’s modulus. Hydrogen as an additive to the DMEM solution improved the preservation effect of osteochondral allograft. The preservation effect of hydrogen occurred in a concentration-dependent manner.