In Vitro Model Alveoli From Photodegradable Templates And Primary Lung Epithelial Cells

摘要

This model has the potential to transform how alveolar development and disease are studied by replacing a flat monolayer with a physiologically relevant 3D structure. Alveolar epithelial cells take on the spherical shape of the microsphere template and maintain a hollow lumen after the template is removed with light. Future work will focus on the ATII cells and the assessment of epithelial polarity, differentiation to type Ⅰ pneumocytes (ATI cells), and presence of tight junctions. Immunostaining for basolateral and apical proteins as well as tight junction proteins will confirm polarity of the epithelial layer. Cell phenotype will be assessed by immunostaining for ATII and ATI specific markers and RT-PCR for phenotype-specific genes,. Further utility of the model will be demonstrated with endothelial cells co-encapsulated within the collagen gels next to the epithelial pre-cysts. In addition, FGF-10 will be added to the media after cyst formation to induce budding of the epithelial layer. Further studies will determine the effect of growth factor location on the directionality of budding.