A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine
A Human 2D Primary Organoid-Derived Epithelial Monolayer Model to Study Host-Pathogen Interaction in the Small Intestine
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DOI：
10.3389/fcimb.2020.00272
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发表时间：
2020-06-09
影响因子：
5.7
通讯作者：
Muncan, Vanesa
中科院分区：
文献类型：
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作者：
Roodsant, Thomas;Navis, Marit;Muncan, Vanesa
肠道类器官是肠上皮干细胞衍生的3D模型，可用于研究肠道病原体与其宿主之间的相互作用。虽然类器官模型已被用于细菌和病毒感染，但这是一个封闭的系统，在没有显微注射或破坏类器官极化的情况下，管腔侧是不可接近的。为了克服这一点，并简化其适用性的跨上皮研究，渗透膜为基础的单层方法是必要的。在本文中，我们展示了一种方法，用于生成单层模型的人胎儿肠极化上皮细胞，充分的特点和验证。近端和远端小肠类器官用于生成2D单层培养物，其特征在于上皮细胞类型、极化、屏障功能和基因表达。此外，还评估了顶端感染肠道病原体肠道病毒A71和单核细胞增生李斯特菌后的病毒复制和细菌移位，并随后监测促炎宿主反应。这种人类2D胎儿肠道单层模型将是研究宿主-病原体相互作用的有价值的工具，并可能减少研究中动物的使用。
Gut organoids are stem cell derived 3D models of the intestinal epithelium that are useful for studying interactions between enteric pathogens and their host. While the organoid model has been used for both bacterial and viral infections, this is a closed system with the luminal side being inaccessible without microinjection or disruption of the organoid polarization. In order to overcome this and simplify their applicability for transepithelial studies, permeable membrane based monolayer approaches are needed. In this paper, we demonstrate a method for generating a monolayer model of the human fetal intestinal polarized epithelium that is fully characterized and validated. Proximal and distal small intestinal organoids were used to generate 2D monolayer cultures, which were characterized with respect to epithelial cell types, polarization, barrier function, and gene expression. In addition, viral replication and bacterial translocation after apical infection with enteric pathogens Enterovirus A71 andListeria monocytogeneswere evaluated, with subsequent monitoring of the pro-inflammatory host response. This human 2D fetal intestinal monolayer model will be a valuable tool to study host-pathogen interactions and potentially reduce the use of animals in research.