Construction Of Tissue Engineering Heart Valves By Human Autologous Cells In Vitro

Objective: To investigate the feasibility of constructing tissue engineering heart valves (TEHV) by human autologous saphenous venous derived endothelial cells (hECs) and myofibroblasts (hMFbs), and human marrow mesenchymal stem cells (hMSCs) for the future clinical application. Methods: hECs, hMFbs and hMSCs were isolated, cultured and expanded in vitro and hMSCs were also induced to differentiate to endothelial cells in vitro (hMSC-ECs) . The phenotypes were identified by immunohistochemistry, and the cell functions were measured; the decellularized porcine scaffolds were made by 0.05%Trypsin and 0.02%EDTA and their histology and biomechanical test were studied; TEHV were constructed under pulsatile flow condition. Results: All the human autologous hECs, hMFbs and hMSCs were isolated, cultured and expanded in vitro with the excellent growth ability especially in hMSCs; Complete decellularization of porcine aortic valve were achieved by 0.05%trypsin and 0.02%EDTA digestion for 24h with intact matrices and biomechanical characteristic preserved; The functions of TEHV, which were affected by both seeding cell types and scaffold preconditioning, improved significantly under pulsatile flow condition. Conclusions: Although the whole function of hMSCs was lower than that of hECs, both human saphenous-derived cells and hMSCs could be the sources of seeding cell for TEHV, and TEHV could be successfully constructed by seeding either hECs or hMSCs on the decellularized porcine scaffolds under pulsatile flow condition in vitro. Further investigations of TEHV are needed on the remodel features in vivo.