Studies have investigated and dismissed the effect of freeze-thaw cycles on both skeletal muscle and on trabecular bone, but have failed to properly address the effects of these storage methods on the integrity of articular cartilage. Preventing cartilage injury is important in minimizing the long term debilitating effects of osteoarthritis. Accurate subfracture injury prediction must take into account the possible effects that freeze thaw cycles may have on the mechanical properties of cartilage tissue. This paper addresses this concern with matched pair testing of various low temperature storage techniques against fresh control groups. Controlled mechanical indention tests were performed on bovine articular cartilage-on-bone specimens to compare stiffness, peak stress, and loading energy of the cartilage. Findings showed that a slow freeze thaw or flash freeze cycle caused cartilage stiffness to decrease by 37% and 31% respectively. Compressive stress at this strain was also lowered by 31% with a single freezing process. These results may be indicative of a weakened extracellular matrix structure caused by the freeze-thaw process. It is still unclear whether these changes in mechanical properties will result in a change in injury susceptibility for articular cartilage.