Cell extraction from porcine aortic valve

AIM To explore methods to remove cellular components from porcine aortic valve,and obtain decellularised scaffolds for tissue engineering of heart valve.METHODS To remove all cells from porcine aortic valve,samples were digested in a 0.1% trypsin solution,followed by multistep detergent-enzymatic process with both hypotonic and hypertonic solutions and ionic detergent.Different detergents(Sodium dodecyl sulfate and sodium deoxycholate,Triton X-100) and enzyme(trypsin) extraction were applied.Specimens were observed grossly and the changes of shrinkage temperature were also studied.Haematoxylin-eosin was performed to confirm the removal of cells and transmission electron microscope was used to observe the integrity of collagen and elastin.RESULTS The cells of porcine aortic valve were removed effectively by Triton X-100 and sodium dodecyl sulfate.Sodium deoxycholate could not effectively remove the cells from the root of aortic valve.The major structural components and shrinkage temperature of decellularised porcine aortic valve by Triton X-100 were well maintained.CONCLUSION Acellular tissue matrix(ACTM) of porcine aortic valve can be successfully obtained through this procedure by Triton X-100.