Decellularization and Recellularization Methods for Avian Lungs: An Alternative Approach for Use in Pulmonary Therapeutics

The shortage of compatible allogeneic organs and an increase in the number of patients requiring long-term lung assist devices while waiting for lung transplantation have motivated scientists to explore alternatives to bioengineer new lungs, including through decellularization and recellularization processes. A novel approach for bioengineering an extracorporeal membrane oxygenator is based on the parenchymal structure of avian lungs which utilizes a cross-current unidirectional flow of air and blood rather than bidirectional airflow, and thus eliminates dead-space ventilation. This provides more efficient gas exchange than mammalian lungs. The novel approach utilized is to decellularize avian lungs and then to recellularize with patient-derived human lung epithelial and vascular endothelial cells with the goal of creating a fully functional structure that can be used as a gas-exchange device. Here, we present avian lung decellularization and recellularization methods for chicken and emu lungs, in order to study both small- and large-scale avian lung models. For decellularization, a detergent-based protocol is utilized, and different techniques are used to validate the de- and recellularization of those lungs, including microscopy, mass spectrometry, and immunohistochemical analyses. For recellularization, techniques for seeding different human lung cell types into the decellularized scaffolds are presented.