Objective: To investigate the feasibility of using acellularized porcine heart valve leaflet as a scaffold for tissue engineering. Methods: The porcine aortic valve leaflets were acellularized by trypsin-EDTA,Triton X-100,RNase and Dnase, and then its biomechanical characteristics was tested and the immunogenicity of acellularized valve leaflet implanted subcutaneous in rats was observed. Results:The valve leaflets were completely removed of the cell components and the construction of matrix was maintained. The biomechanical characteristics of the acellular valve leaflets and the fresh ones were almost similar. The immunogenicity of the acellularized valve leaflets was alleviated significantly compared with that of fresh ones. Conclusion:Acellularized porcine aortic valve leaflets can be applied to develop tissue engineering heart valve as a scaffold.