Several deficiencies in currently available right ventricular valved conduits make them problematic for use in infants and children. A solution would be to develop a tissue-engineered valved conduit containing autologous cells. A method was devised to produce a decellularized bovine matrix scaffold for developing a tissue-engineered right ventricular valved conduit. Fresh bovine jugular veins were treated with sodium deoxycholate and Triton X-100. The major structural proteins of the fresh and decellularized jugular venous valves and vessel walls were detected by histological methods. Thickness, water absorption rate, water maintenance rate, disruption strength, and extensibility were determined. Circumferential and radial specimens of valves and vessel walls were subjected to tensile testing. Histological analysis showed that no cell fragments were retained within the decellularized matrix scaffold and the major structural proteins had been retained intact. There were no significant differences in thickness, rates of absorption and maintenance of water, disruption strength, and extensibility between the decellularized and fresh veins. It was concluded that this treatment can successfully remove cellular components while maintaining the major structural components and the histological and biological properties of bovine jugular veins.