GSE158035
Single-Cell RNA Sequencing Pinpoints Exocrine Bipotentiality of Human Pancreatic Organoids
Objective: Despite organoid formation and characterization from different tissues have been extensively studied, pancreas remains one among the least investigated organs. Here we proposed a multifaced characterization of cell types present in human pancreatic organoids (hPO) and starting from their progenitor we described their maturation and their lineage relationships. Design: We performed unbiased transcriptome-wide scRNA-seq analysis on 3187 cells of human pancreatic organoids obtained from 3 healthy islet-depleted tissue. To support our results, we strengthened these findings with proteomic and qRT-PCR analyses performed during the long-term culture of hPO. Results: First, we described hPO focusing on their morphology and long-term culture molecular aspects and showing a well-defined epithelial organization supported by a genetic stability through all cell passages. Successively, based on single-cell RNA sequencing technology, we defined all cell types present in hPO confirming that our pancreatic organoid methodology is highly reproducible. Moreover, exploiting the latest bioinformatic tools and comparing our data with recently published ones we reshaped the conventional theory of univocal ductal nature of hPO. Finally, we defined a new strategy to sort a refined multipotent progenitor from hPO and we gave a new definition of pancreatic progenitor demonstrating its capacity to renew the hPO stem population while maintaining its heterogeneity. Conclusions: Taken together, our results give a new definition of human pancreatic organoids, which reveals the complex nature of these structures and provides a strong basis to pave the way for future study in the pancreatic field. Overall design: Single cell RNA-seq profile of human pancreatic organoids obtained from 3 healthy islet-depleted tissue