There are several types of bioreactors currently available for the culture of orthopaedic tissue engineered constructs. These vary from the simple to the complex in design and culture. Preparation of samples for bioreactors varies depending on the system being used. This chapter presents data and describes tried and tested methodologies for the preparation of 3D samples for a Rotatory Synthecon Bioreactor (Cellon), a plate shaker, a perfusion system, and a Bose Electroforce Systems Biodynamic Instrument for the in vitro culture of bone and ligament tissue.