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Developing an ex-vivo organotypic co-culture system to model airway epithelial cell-chondrocyte interactions during tracheal development

In this project, we study the effects of airway epithelial cell (AEC)-chondrocyte co-culture on hyaline cartilage formation and ECM deposition as a model of tracheal development.

Our group has an active research interest in the role of AECs in the coordination of airway remodeling and airway inflammation associated with alteration in the ECM during lung disease. Our lab has previously developed an ex vivo AEC-human lung fibroblast (HLF) co-culture model system to investigate the role of AEC-HLF crosstalk on ECM deposition and composition. In this project, we capitalize on the existing expertise with AEC-HLF co-culture model system to evaluate ECM changes to establish a new model of AEC-chondrocyte crosstalk to evaluate alterations in hyaline cartilage formation and ECM composition.

Using primary mesenchymal stromal cells (MSCs) differentiated into chondrocytes in ex vivo organoids, we will determine the effect of AEC-chondrocyte co-culture on the formation of hyaline cartilage and ECM deposition by co-cultured chondrocytes.

A primary goal of ours is to differentiate chondrocytes from MSCs obtained from pediatric donors with known FGFR2 craniosynostosis syndromes to be utilized in the AEC-chondrocyte co-cultures in collaboration with the Eldredge and Dahl Labs. This will allow us to determine the effect of FGFR2 gain of function signaling on AEC differentiation and ciliogenesis compared to co-culture with chondrocytes differentiated from MSCs obtained from healthy donors.

We will also characterize the differences in the hyaline cartilage and ECM deposition by chondrocytes with a FGFR2 gain of function mutation co-cultured with AECs compared to co-cultured chondrocytes derived from unaffected donors.

These studies will give us insight to into the altered regulation of tracheal cartilage deposition that may underline the development of TCS.

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