Single-Cell RNA Sequencing Pinpoints Exocrine Bipotentiality of Human Pancreatic Organoids

Human organoids have been proposed to be powerful tools mimicking the physiopathological processes of the organs of origin. Recently, human pancreatic organoids (hPOs) have gained increasing attention due to potential regenerative medicine applications. However, the cellular components of these three-dimensional structures have not been defined precisely. In this work, we finely characterized hPOs, focusing first on morphology and identity-defining molecular features under long-term culture conditions. We observed consistent cell growth, accompanied by genetic stability, expression of pancreatic markers, and epithelial organization. Next, we identified hPO cell types using single-cell RNA sequencing. Exploiting cutting-edge bioinformatics pipelines, we reshaped the conventional theory of the univocal ductal nature of hPOs into a ductal and acinar bipotentiality. Finally, we proposed a new definition of the multipotent pancreatic hPO progenitors using a refined sorting strategy, with demonstration of self-renewal and differentiation capabilities, thus providing a strong foundation for future studies of pancreatic health and disease.