Department of Biomedical and Clinical Sciences, Faculty of Health Science, Linköping University, Linköping, Sweden.
Department of Biomedical and Clinical Sciences, Faculty of Health Science, Linköping University, Linköping, Sweden; Wallenberg Centre for Molecular Medicine, Linköping University, Linköping, Sweden.
Methods Cell Biol. 2023;179:21-38. doi: 10.1016/bs.mcb.2022.12.005. Epub 2023 May 16.
Multicellular organisms need epithelial barriers to remain compartmentalized and protected from external influences. Although much progress has been made in understanding barrier integrity disruption in Celiac disease (CD), the regulatory and genetic mechanisms underlying the increased intestinal epithelial flux are still unknown. As we learn more about the regulation of permeability in homeostasis and pathogenesis, we will be able to develop strategies to strengthen the epithelial barrier function in intestinal disorders, including CD. For this purpose, Ussing chambers are increasingly used in native tissue, such as gut mucosa or cell monolayers, to assess the integrity of the barrier. In particular, the Ussing chambers allow the measurement of paracellular and transcellular parameters of CD small intestinal biopsies under physiologically specific conditions. In diverse types of diseases, this method is commonly used to determine epithelial barrier defects, but its application to CD has not yet been widely expanded. To provide a great model of barrier ex vivo studies in CD, we facilitate a standard protocol to measure paracellular and transcellular permeability using the Ussing chamber.
多细胞生物需要上皮屏障来保持分隔和免受外部影响。尽管在理解乳糜泻 (CD) 中屏障完整性破坏方面已经取得了很大进展，但增加的肠道上皮细胞通量的调节和遗传机制仍不清楚。随着我们对稳态和发病机制中通透性调节的了解不断加深，我们将能够制定策略来增强包括 CD 在内的肠道疾病中的上皮屏障功能。为此，Ussing 室越来越多地用于评估天然组织（如肠道黏膜或细胞单层）的屏障完整性。特别是，Ussing 室允许在生理特异性条件下测量 CD 小肠活检的旁细胞和跨细胞参数。在多种类型的疾病中，该方法通常用于确定上皮屏障缺陷，但尚未广泛应用于 CD。为了提供 CD 体外屏障研究的良好模型，我们提供了使用 Ussing 室测量旁细胞和跨细胞通透性的标准方案。
