Celiac Disease Research Center, Faculty of Medicine and Health Technology, Tampere University, Tampere University Hospital, Tampere, Finland.
Faculty of Medicine and Health Technology, Tampere University, Tampere, Finland.
Methods Cell Biol. 2023;179:173-193. doi: 10.1016/bs.mcb.2023.01.008. Epub 2023 Feb 24.
Celiac disease pathogenesis, in addition to immune cell component, encompasses pathogenic events also in the duodenal epithelium. In celiac disease patients, exposure to dietary gluten induces drastic changes in epithelial differentiation and elicit cellular response to inflammatory cytokines. The autoantigen in celiac disease, transglutaminase 2 (TG2) enzyme, has been also suggested to play its pathogenic gliadin deamidation event in the intestinal epithelium. Therefore in vitro epithelial cell-line models have been exploited in the past to study these pathogenic mechanisms, but they are hampered by their simplistic nature lacking proper cell-type composition and intestinal environ. Moreover, these cell models harbor many cancer-related mutations in tumor suppressor genes making them unsuitable for studying cell differentiation. Intestinal organoids provide a near-native epithelial cell model to study pathogenic agents and mechanisms related to celiac disease. Here we describe protocols to initiate and maintain celiac patient-derived organoid cultures and how to grow them in alternative ways allowing their exploitation in different kind of experiments.
乳糜泻的发病机制除了免疫细胞成分外，还包括十二指肠上皮中的致病事件。在乳糜泻患者中，膳食 gluten 的暴露会引起上皮分化的剧烈变化，并引起对炎症细胞因子的细胞反应。乳糜泻中的自身抗原，转谷氨酰胺酶 2（TG2）酶，也被认为在肠上皮中发挥其致病的麦胶去酰胺作用。因此，过去曾利用体外上皮细胞系模型来研究这些致病机制，但它们受到自身简单性质的限制，缺乏适当的细胞类型组成和肠道环境。此外，这些细胞模型中存在许多肿瘤抑制基因的癌症相关突变，使它们不适合研究细胞分化。肠类器官提供了一种接近天然的上皮细胞模型，可用于研究与乳糜泻相关的致病因子和机制。在这里，我们描述了启动和维持乳糜泻患者来源的类器官培养的方案，以及如何以不同的方式生长它们，从而允许在不同类型的实验中利用它们。
