Proper in vitro models to evaluate the performance of relevant tissue engineered constructs are still a major demand of the field. The use of in vitro platforms presents obvious ethical and cost advantages over in vivo models. In vitro models can also offer important scientific benefits in the study of biological mechanisms of action focusing on isolated variables/effects. Different 2D and 3D cell culturing systems, static (e.g., tissue culture polysterene with or without transwell) and dynamic (e.g., bioreactors), have been proposed along the years. However, the lack of suitable physical structures capable of supporting the generation of multilayered and complex tissues has been hampering the field of tissue engineering [1]. Among the challenges to attain multilayered structures, the creation of a continuous gradient that allow a smooth interface formation between layers is one of the most demanding [2]. In addition, improved cell culturing conditions in a 3D environment are still to be achieved and one of the biggest problems to be overcome. In this sense, this work refers to the development of a rotational dual chamber bioreactor adapted for cell culture in multi-layered scaffolds, providing an enhanced cellular homogenization inside of the layered 3D structure. Ultimately the developed system will allow creating an osteochondral 3D in vitro model.