A Fully Automated High-Throughput Bioreactor System for Lung Regeneration

The careful study of cell-based lung repair and regeneration ex vivo may one day provide us with the necessary tools to create a patient-derived alternative to cadaveric donor lungs for transplantation. Many parameters must be monitored and optimized to advance this aim. The use of rat lungs as a small-scale model for lung regeneration is an efficient way to develop the key improvements required for optimal tissue repair and regeneration. In this study, we report the use of a novel high-throughput, automated, multichannel lung bioreactor system, which allows for culture and analysis of rodent scale isolated lungs. In this model, five decellularized rat lungs can be repopulated with human primary endothelial and epithelial cells, cultured under varying perfusion and ventilation parameters in parallel, and analyzed for standardized endpoints. As a proof of principle, we report a multiphase organ culture protocol, which achieves consistent tissue regeneration across lungs at multiple points during culture, and further promotes overall tissue maturation through the application of lung ventilation. Impact Statement This work presents methods for ex vivo lung recellularization and biomimetic culture in a high-throughput and consistent manner. These methods allow for the testing of multiple variables, all of which are simultaneously controlled and monitored on a single fully automated pump system, and subsequent assessment of both epithelial and endothelial repair and tissue regeneration. This system provides a controlled environment for tissue repair, wherein key variables can be modified, monitored, reproduced, and optimized to advance the goal of ex vivo tissue regeneration based on native organ scaffolds.