Esophageal progenitor cell plasticity; a 3D organotypic model of epithelial regeneration

Epithelial stem cells are capable of remarkable plasticity, facilitating efficient regenerative responses following damage. Here, we present a versatile model of ectopic niche regeneration that provides evidence of a switch in epithelial stem cell identity as a response to foreign microenvironmental signals. A combination of 3D organotypic cultures, 3D confocal imaging and Next-Generation Sequencing reveal events of epithelial plasticity during the reepithelialization of denuded stroma. In this study we perform a temporal analysis that provides new insights on the mechanisms driving cell fate flexibility in adult epithelial stem cells. Single cell RNA sequencing of oesophageal epithelial cells cultured in a 3D organotypic system that recapitulates wound healing. Cells growing over their native stroma were compared to those growing over the foreign stroma of the dermis. The temporality of the process was captured by sequencing cells at day 3 and 10 into the regeneration process. For each of the four conditions, at least 3 biological replicates were performed.