Osteochondral defects (i.e., those that affect both the articular cartilage and underlying subchondral bone) are often associated with mechanical instability of the joint, and therefore with the risk of inducing osteoarthritic degenerative changes. The in vitro fabrication of osteochondral grafts of predefined size and shape, starting from autologous cells combined with three-dimensional porous biomaterials, is a promising approach for the treatment of osteochondral defects. However, the quality of ex vivo generated cartilage and bone-like tissues is currently restricted by a limited understanding of the regulatory role of physicochemical culture parameters on tissue development. By allowing reproducible and controlled changes in specific biochemical and biomechanical factors, bioreactor systems provide the technological means to reveal fundamental mechanisms of cell function in a three-dimensional environment and the potential to improve the quality of engineered tissues. In addition, by automating and standardizing the manufacturing process in controlled closed systems, bioreactors could reduce production costs and thus facilitate broader clinical impact of engineered osteochondral grafts.