Relevant human in vitro culture models can contribute to a reduction in animal use for respiratory research and generate data that can be better translated to patients. In this thesis various... Show moreRelevant human in vitro culture models can contribute to a reduction in animal use for respiratory research and generate data that can be better translated to patients. In this thesis various improvements of different current human lung in vitro models were explored. Human induced pluripotent stem cells (hiPSC) were used to generate alveolar type 2 (AEC2)-like cells, that were subsequently cultured at the air-liquid interface (ALI) to create a model of alveolar wound healing at the ALI. Furthermore, a method to utilize organoids was developed for expansion of cells from human lungs to obtain sufficient numbers to reliably establish epithelial cultures. This was done for AEC2 isolated from resected lung tissue, as well as for cell populations from broncho-alveolar lavage fluid from adults and tracheal aspirates from preterm new-borns. Finally, the cellular complexity of the in vitro cultures was expanded by introducing polarized macrophages to the airway ALI cultures. Using these cultures, the cross-talk between macrophages and airway was studied in relation to wound repair. Collectively, the methods that have been developed are expected to contribute to contribute to the use of patient- and disease relevant models in respiratory research. Show less