Abstract: Objective : Construction of Tissue engineering heart valves in vitro. Methods: firstly, comparison of different methods of obtaining acellularized valve xenografts. Specimens of porcine aortic roots were treated with either Trypsin-sodium deoxycholate（SD）or sodium-dodecyl-sulfate (SDS) or Triton-X100-SD with specific concentrations. Tissue samples were then processed for scanning electron microscopy and Hematoxylin- Eosin and Masson’s trichrome. Observe cell-scaffold under light microscope and scanning electron microscopy. Then, bone marrow mesenchymal stem cells (MSCs) were harvested , cultured and expended and reseeded on the acellularized valve xenografts under static conditions for 7 days .Results: compared with the treatments of Trypsin -SD and SDS, Triton-SD achieved both complete decellularization and preservation of the matrix structure. Cell-scaffold after implantation formed three dimensional tissue, scanning electron microscopy show bone MSCs can grow on the surface of Scaffold. Conclusion: Techniques of decellularization are highly variable in efficiency and matrix preservation and was best achieved in our study with Triton-SD. Bone MSCs are good sources of seeding cells which can adhesion and grow well to decelluarized porcine heart valve in vitro environment.

Key words: Tissue engineering Heart valves, Bone marrow mesenchymal stem cells (MSCS), Triton, Decelluarized porcine heart valve