Automation of Pressure Control Improves Whole Porcine Heart Decellularization

作者：Momtahan Nima; Poornejad Nafiseh; Struk Jeremy A; Castleton Arthur A; Herrod Brenden J; Vance Brady R; Eatough Jordan P; Roeder Beverly L; Reynolds Paul R; Cook Alonzo D^*

来源：Tissue Engineering - Part C: Methods , 2015, 21(11): 1148-1161.

DOI：10.1089/ten.tec.2014.0709

Whole heart decellularization combined with patient-specific cells may prove to be an extremely valuable approach to engineer new hearts. Mild detergents are commonly used in the decellularization process, but are known to denature and solubilize key proteins and growth factors and can therefore be destructive to the extracellular matrix (ECM) during the decellularization process. In this study, the decellularization of porcine hearts was accomplished in 24h with only 6h of sodium dodecyl sulfate exposure and 98% DNA removal. Automatically controlling the pressure during decellularization reduced the detergent exposure time while still completely removing immunogenic cell debris. Stimulation of macrophages was greatly reduced when comparing native tissue samples to the processed ECM. Complete cell removal was confirmed by analysis of DNA content. General collagen and elastin preservation was demonstrated. Glycosaminoglycans and collagen quantification both showed no significant differences in content after decellularization. The compression elastic modulus of the ECM after decellularization was lower than native at low strains, but there was no significant difference at high strains. Polyurethane casts of the vasculature of native and decellularized hearts demonstrated that the microvasculature network was preserved after decellularization. A static blood thrombosis assay using bovine blood was also developed. Finally, the recellularization potential of the ECM samples was demonstrated by reseeding cardiac fibroblasts and endothelial cells on the myocardium and endocardium samples.

• 出版日期2015-11-1