Organoids for modelling the physiology and pathology of gastrointestinal tissues are constrained by a poorly accessible lumen. Here we report the development and applicability of bilaterally accessible organoid-derived patterned epithelial monolayers that allow the independent manipulation of their apical and basal sides. We constructed gastric, small-intestinal, caecal and colonic epithelial models that faithfully reproduced their respective tissue geometries and that exhibited stem cell regionalization and transcriptional resemblance to in vivo epithelia. The models’ enhanced observability allowed single-cell tracking and studies of the motility of cells in immersion culture and at the air–liquid interface. Models mimicking infection of the caecal epithelium by the parasite Trichuris muris allowed us to live image syncytial tunnel formation. The enhanced observability of bilaterally accessible organoid-derived gastrointestinal tissue will facilitate the study of the dynamics of epithelial cells and their interactions with pathogens.

用于模拟胃肠道组织生理学和病理学的类器官受到难以接近的管腔的限制。在这里，我们报告了双侧可及的类器官衍生的图案上皮单层的开发和适用性，这些单层允许独立操作其顶端和基底侧。我们构建了胃、小肠、盲肠和结肠上皮模型，这些模型忠实地再现了它们各自的组织几何形状，并表现出与体内上皮细胞的干细胞区域化和转录相似性。这些模型增强的可观察性允许单细胞跟踪和研究浸没培养物和气液界面中细胞的运动。模拟寄生虫 Trichuris muris 感染盲肠上皮的模型使我们能够实时成像合胞体隧道的形成。双侧可接近的类器官来源的胃肠道组织的增强可观察性将有助于研究上皮细胞的动力学及其与病原体的相互作用。