Characterization of human pancreatic tissue and organoids reveals subpopulation of intercalated duct cells with progenitor traits

Pancreatic duct organoids have emerged as a valuable tool for investigating pancreas biology and physiology through in vitro modeling. Despite their widespread use, a comprehensive understanding of the cellular heterogeneity of organoids and their underlying tissue remains elusive. Here we performed single-cell transcriptomic profiling of human adult pancreatic exocrine tissue and their derived organoids, revealing a gradient of keratin expression within the ductal network that clustered cells into two distinct low- and high-keratin subpopulations. Keratin-high cells display a higher capacity to form organoids while keratin-low cells express stemness associated markers. Finally, comparative gene expression analysis between ductal tissue and derived organoids revealed a partial recapitulation of the in vivo heterogeneity. These findings further our knowledge on pancreatic duct organoids and pave the way for future investigations into the heterogeneity of the pancreatic ductal syste Overall design: Pancreatic islet-depleted tissue of 5 donors and 2 organoid lines was sequenced utilizing the SORT-seq platform. Samples of 2 donors were additionally collected using Fluorescence activated cell sorting based on expression of cell surface marker GP2 to study both positive and negative populations.

胰腺导管类器官（pancreatic duct organoids）已成为通过体外建模研究胰腺生物学与生理学的宝贵工具。尽管其应用已十分广泛，但学界对类器官及其来源组织的细胞异质性仍缺乏全面认知。本研究对人类成人胰腺外分泌组织及其衍生类器官开展了单细胞转录组谱分析，揭示了导管网络内的角蛋白表达梯度，该梯度将细胞聚类为两个截然不同的角蛋白低表达与高表达亚群。角蛋白高表达细胞形成类器官的能力更强，而角蛋白低表达细胞则表达干细胞干性相关标志物。最终，通过对导管组织与衍生类器官开展比较基因表达分析，本研究发现类器官部分重现了体内的细胞异质性。上述发现深化了我们对胰腺导管类器官的认知，也为未来针对胰腺导管系统异质性的研究铺平了道路。实验整体设计：本研究利用SORT-seq测序平台，对5名供者的去除胰岛的胰腺组织以及2株类器官系进行了测序。此外，研究人员基于细胞表面标志物GP2的表达，通过荧光激活细胞分选（Fluorescence activated cell sorting）收集了2名供者的样本，以分别研究阳性与阴性细胞群。