Decellularization of porcine whole lung to obtain a clinical-scale bioengineered scaffold

Whole-organ engineering is emerging as an alternative source for xenotransplantation in end-stage diseases. Utilization of decellularized whole lung scaffolds created by detergent perfusion is an effective strategy for organ replacement. In the current study, we attempted to decellularize porcine whole lungs to generate an optimal and reproducible decellularized matrix for future clinical use. Porcine whole lungs were decellularized via perfusion of various detergents (sodium dodecyl sulfate (SDS)/Triton X-100, sodium lauryl ether sulfate (SLES)/Triton X-100, dextrose/SDS/Triton X-100 and dextrose/SLES/Triton X-100) through the pulmonary artery and bronchus of the lung. The decellularized scaffolds were evaluated for decellularization efficiency, extracellular matrix (ECM) component preservation, xenoantigen removal and compatibility. The resulting lung scaffolds obtained from treatment with the dextrose/SLES/Triton X-100 cocktail showed minimal residual cellular components and xenoantigens, including DNA and protein, and good preservation of ECM composition. Evaluation of the porcine lung ECM by specific staining and immunofluorescence confirmed that the three-dimensional ultrastructure of the ECM was noticeably preserved in the SLES-treated groups. In addition, the decellularized lung scaffolds originating from the dextrose/SLES/Triton X-100 cocktail supported cell adhesion and growth. In summary, the novel detergent SLES alleviated the damage to retain a better-preserved ECM than SDS. Sequential Triton X-100 perfusion eliminated SLES. Moreover, performing dextrose perfusion in advance further protected scaffold components, especially collagen. We developed an optimal dextrose/SLES/Triton X-100 cocktail method that can be used for the decellularization of porcine whole lung to obtain a clinical-scale bioengineered scaffold.