Clinical Physiology/Nutritional Medicine, Charité - Universitätsmedizin Berlin, CBF, Berlin, Germany.
Department of Infectious Diseases, Unit 16 Mycotic and Parasitic Agents and Mycobacteria, Robert Koch-Institute, Berlin, Germany.
Ann N Y Acad Sci. 2022 Sep;1515(1):155-167. doi: 10.1111/nyas.14804. Epub 2022 Jun 6.
Usually, duodenal barriers are investigated using intestinal cell lines like Caco-2, which in contrast to native tissue are limited in cell-type representation. Organoids can consist of all intestinal cell types and are supposed to better reflect the in vivo situation. Growing three-dimensionally, with the apical side facing the lumen, application of typical physiological techniques to analyze the barrier is difficult. Organoid-derived monolayers (ODMs) were developed to overcome this. After optimizing culturing conditions, ODMs were characterized and compared to Caco-2 and duodenal tissue. Tight junction composition and appearance were analyzed, and electrophysiological barrier properties, like paracellular and transcellular barrier function and macromolecule permeability, were evaluated. Furthermore, transcriptomic data were analyzed. ODMs had tight junction protein expression and paracellular barrier properties much more resembling the originating tissue than Caco-2. Transcellular barrier was similar between ODMs and native tissue but was increased in Caco-2. Transcriptomic data showed that Caco-2 expressed fewer solute carriers than ODMs and native tissue. In conclusion, while Caco-2 cells differ mostly in transcellular properties, ODMs reflect trans- and paracellular properties of the originating tissue. If cultured under optimized conditions, ODMs possess reproducible functionality, and the variety of different cell types makes them a suitable model for human tissue-specific investigations.
通常，十二指肠屏障是使用肠细胞系（如 Caco-2）进行研究的，与天然组织相比，这些细胞系在细胞类型的代表性上受到限制。类器官可以包含所有的肠细胞类型，并且应该能够更好地反映体内情况。由于它们是三维生长的，顶端朝向腔，因此应用典型的生理技术来分析屏障是困难的。类器官衍生的单层（ODM）的开发就是为了克服这一困难。在优化培养条件后，对 ODM 进行了表征，并与 Caco-2 和十二指肠组织进行了比较。分析了紧密连接的组成和形态，评估了电生理屏障特性，如旁细胞和跨细胞屏障功能以及大分子通透性。此外，还分析了转录组数据。ODM 具有紧密连接蛋白表达和旁细胞屏障特性，与起源组织比 Caco-2 更相似。跨细胞屏障在 ODM 和天然组织之间相似，但在 Caco-2 中增加。转录组数据显示，Caco-2 表达的溶质载体比 ODM 和天然组织少。总之，虽然 Caco-2 细胞在跨细胞特性上差异最大，但 ODM 反映了起源组织的跨细胞和旁细胞特性。如果在优化的条件下培养，ODM 具有可重复的功能，并且不同细胞类型的多样性使它们成为适合人类组织特异性研究的模型。
