This project focuses on a developmental candidate that is an airway construct, composed of a customized tissue engineered biologic scaffold, populated with cells from the recipient (mesenchymal and epithelial). The construct is a decellularized donor section of trachea, based on implants used in humans in compassionate use cases (adults and children). The studies, to date, have focused on optimized techniques for collecting and characterizing cells, tracheal decellularization techniques, and tracheal recellularization strategies. A summary of achievements includes the following: (1) Cells: comparable marker expression and tri-lineage differentiation potential shown, and no differences in phenotypic or functional properties with transduction to express reporter genes for imaging confirmed; (2) Decellularization: decellularization standard operating procedures (SOP) identified, decellularized trachea shown to meet all success criteria, and biomechanical testing optimized; (3) Recellularization: compared bioreactor designs and identified a recellularization SOP, and optimized conditions for external and intraluminal seeding. In Year 1 we have made significant progress with new and innovative approaches to meet our milestones and goals.