Establishment of a high-resolution 3D modeling system for studying pancreatic epithelial cell biology in vitro
Establishment of a high-resolution 3D modeling system for studying pancreatic epithelial cell biology in vitro
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DOI：
10.1016/j.molmet.2019.09.005
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发表时间：
2019-12-01
影响因子：
8.1
通讯作者：
Lickert, Heiko
中科院分区：
文献类型：
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作者：
Bakhti, Mostafa;Scheibner, Katharina;Lickert, Heiko
目的：基础研究从实验室到临床的转化依赖于对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的更好理解。因此，建立小鼠和人类胰腺发育的离体模型系统，不仅有助于理解进化保守的分化和形态发生机制，而且有助于理解疾病的病理机制和设计组织工程策略。在这里，我们建立了一种简单、可重复的小鼠和人胰腺祖细胞（PP）基于Matrigel的三维（3D）囊肿培养模型系统，以离体研究胰腺上皮形成和内分泌发生。此外，我们重新分析了以前报道的单细胞RNA测序（scRNA-seq）的小鼠和人类胰腺谱系，以获得一个全面的图片的差异表达的关键转录因子（TF），细胞-细胞粘附分子和细胞极性组件在PPs在endocrinogenesis.Results：我们产生了小鼠和人类极化胰腺上皮囊肿来自PPs。该系统允许以动态时间分辨方式监测细胞和亚细胞分辨率中胰腺上皮极性和管腔形成的建立。此外，小鼠和人胰腺囊肿都能够向内分泌命运分化。该分化系统与scRNA-seq分析一起揭示了内分泌differentiation.Conclusions过程中顶端-基底极性以及紧密和粘附连接的变化：我们已经建立了一个简单的3D胰腺囊肿培养系统，该系统允许在机械水平上时空分辨细胞和亚细胞过程，否则在体内是不可能的。(C)2019年，任作家。由爱思唯尔有限公司出版。
Objective: 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.Methods: 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.Results: 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.Conclusions: 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. (C) 2019 The Authors. Published by Elsevier GmbH.