LAUSR

More than one billion people worldwide suffer from congenital and chronic respiratory diseases, leading to high rates in mortality and morbidity. The unpredictability of disease progression and responsiveness towards treatment requires sophisticated experimental models to better mimic the patient-specific biology of the airways. Currently, the best reproducible in vitro airway system is the air-liquid interface (ALI) method, a static, air-exposed airway epithelial layer. However, this model lacks the cellular diversity and 3D structure of the in vivo microenvironment, the flow dynamics and the mechanical stretch of the human lung. To address these issues, we aim to stepwise develop a dynamic and reproducible three-dimensional model of the human airway by co-culturing human primary cells and/or induced pluripotent stem cell (iPSC)-derived lung epithelial cells. First, we optimized media culture conditions for the co-cultures of human primary lung endothelial cells (hLECs) with human primary bronchial epithelial cells (hPBECs) and human primary lung fibroblasts (hLFs). We will show the co-culture medium that best promotes survival, proliferation and differentiation of hPBECs to mucociliary cells. Next, we increased the complexity of the ALI culture system by seeding hPBECs on top of hydrogels containing hLFs, and seeding hLECs at the basal side of the insert membrane. Transdifferentiation of the different cell types will also be evaluated to prevent cells from losing their phenotype and being under stress in the co-culture model. These complex cultures will subsequently be subjected to dynamic flow to investigate the effect on the viability and differentiation potential of the various cell types.