Human Lung Organoid Culture in Alginate With and Without Matrigel to Model Development and Disease

Human lung organoids (HLOs) are enabling the study of human lung development and disease by modeling native organ tissue structure, cellular composition, and cellular organization. In this report, we demonstrate that HLOs derived from human pluripotent stem cells cultured in alginate, a fully defined nonanimal product substrate, exhibit enhanced cellular differentiation compared with HLOs cultured in the commercially available Matrigel. More specifically, we observed an earlier onset and increase in the number of multiciliated cells, along with mucus producing MUC5AC+ goblet-like cells that were not observed in HLOs cultured in Matrigel. The epithelium in alginate-grown HLOs was organized in a pseudostratified epithelium with airway basal cells lining the basal lamina, but with the apical surface of cells on the exterior of the organoid. We further observed that HLOs cultured in Matrigel exhibited mesenchymal overgrowth that was not present in alginate cultures. The containment of the mesenchyme within HLOs in alginate enabled modeling of key features of idiopathic pulmonary fibrosis (IPF) by treatment with transforming growth factor β (TGFβ). TGFβ treatment resulted in morphological changes including an increase in mesenchymal growth, increased expression of IPF markers, and decreased numbers of alveolar-like cells. This culture system provides a model to study the interaction of the mesenchyme with the epithelium during lung development and diseased states such as IPF. Human lung organoids (HLOs) can potentially be used as model systems for diseases of the lung; however, they are currently limited by the ability of existing culture systems to support functional organoids. In this study, we describe an alginate culture system that supports the development of HLOs from embryonic stem cells. Organoids grown in alginate both model key aspects of lung physiology and display hallmarks of fibrosis when stimulated with transforming growth factor β. This model system could potentially be used to screen treatments for fibrotic lungs and provide crucial insights into lung biology.