Abstract
Tissue engineering could now be considered as one of the main spheres of the medical basic science and shows a dramatical increase in number of publications. Decellularization is one of the tissue engineering methods that results in non-immunogenic tissue matrix as a basis for future tissue construction. The comparison of multiple decellularization methods is nowadays quite complex as far as there is no standardized approach to the decellularization quality control. 19 different detergent-based decellularizations protocol were analyzed for treatment of whole ovine mitral valves (n=78). The decellularization quality control included three levels: routine light microscopy (hematoxylin-eosin staining), immunofluorescent analysis for DNA (DAPI staining) and immunofluorescent analysis for ?-GAL (Lectin I Isolectin B4 staining). The best decellularization results were shown by the protocol using detergents (sodium dodecyl sulfate and sodium deoxycholate) in rising concentrations with reducing agent (?-mercaptoethanol). The routine light microscopy wasshown to be the least sensitive method, opposite to immunofluorescent analysis for membrane antigen ?-GAL. The developed three level decellularization quality control approach was shown to be effective and could be used for non-mammalian tissues.