Institute of Diabetes and Regeneration Research, Helmholtz Zentrum München, D-85764, Neuherberg, Germany; German Center for Diabetes Research (DZD), D-85764, Neuherberg, Germany; Institute of Stem Cell Research, Helmholtz Zentrum München, D-85764, Neuherberg, Germany.
Institute of Diabetes and Regeneration Research, Helmholtz Zentrum München, D-85764, Neuherberg, Germany; German Center for Diabetes Research (DZD), D-85764, Neuherberg, Germany; Institute of Stem Cell Research, Helmholtz Zentrum München, D-85764, Neuherberg, Germany; Technical University of Munich, School of Medicine, Munich, Germany.
Mol Metab. 2019 Dec;30:16-29. doi: 10.1016/j.molmet.2019.09.005. Epub 2019 Sep 24.
Translation of basic research from bench-to-bedside relies on a better understanding of similarities and differences between mouse and human cell biology, tissue formation, and organogenesis. Thus, establishing ex vivo modeling systems of mouse and human pancreas development will help not only to understand evolutionary conserved mechanisms of differentiation and morphogenesis but also to understand pathomechanisms of disease and design strategies for tissue engineering.
Here, we established a simple and reproducible Matrigel-based three-dimensional (3D) cyst culture model system of mouse and human pancreatic progenitors (PPs) to study pancreatic epithelialization and endocrinogenesis ex vivo. In addition, we reanalyzed previously reported single-cell RNA sequencing (scRNA-seq) of mouse and human pancreatic lineages to obtain a comprehensive picture of differential expression of key transcription factors (TFs), cell-cell adhesion molecules and cell polarity components in PPs during endocrinogenesis.
We generated mouse and human polarized pancreatic epithelial cysts derived from PPs. This system allowed to monitor establishment of pancreatic epithelial polarity and lumen formation in cellular and sub-cellular resolution in a dynamic time-resolved fashion. Furthermore, both mouse and human pancreatic cysts were able to differentiate towards the endocrine fate. This differentiation system together with scRNA-seq analysis revealed how apical-basal polarity and tight and adherens junctions change during endocrine differentiation.
We have established a simple 3D pancreatic cyst culture system that allows to tempo-spatial resolve cellular and subcellular processes on the mechanistical level, which is otherwise not possible in vivo.
基础研究从实验室到临床的转化依赖于更好地理解小鼠和人类细胞生物学、组织形成和器官发生之间的相似性和差异性。因此，建立小鼠和人胰腺发育的体外模型系统不仅有助于理解分化和形态发生的进化保守机制，也有助于理解疾病的病理机制并设计组织工程策略。
我们建立了一种简单且可重复的基于基质胶的三维（3D）小鼠和人胰腺祖细胞（PP）类器官培养模型系统，用于体外研究胰腺上皮化和内分泌发生。此外，我们重新分析了先前报道的小鼠和人胰腺谱系的单细胞 RNA 测序（scRNA-seq）数据，以全面了解在内分泌发生过程中 PP 中转录因子（TFs）、细胞间黏附分子和细胞极性成分的差异表达。
我们生成了源自 PP 的小鼠和人极化胰腺上皮类器官。该系统允许以动态时间分辨的方式在细胞和亚细胞分辨率监测胰腺上皮极性的建立和腔形成。此外，小鼠和人胰腺类器官均能向内分泌命运分化。该分化系统结合 scRNA-seq 分析揭示了在上皮向内分泌分化过程中顶端-基底极性和紧密连接及黏附连接是如何改变的。
我们建立了一种简单的 3D 胰腺类器官培养系统，能够在机制水平上实时解析细胞和亚细胞过程，而这在体内是不可能实现的。
