Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), Tokyo, Japan.
Department of Research and Development for Organoids, Juntendo University School of Medicine, Tokyo, Japan.
Nat Protoc. 2022 Mar;17(3):649-671. doi: 10.1038/s41596-021-00658-3. Epub 2022 Feb 2.
Intestinal organoids are fundamental in vitro tools that have enabled new research opportunities in intestinal stem cell research. Organoids can also be transplanted in vivo, which enables them to probe stem cell potential and be used for disease modeling and as a preclinical tool in regenerative medicine. Here we describe in detail how to orthotopically transplant epithelial organoids into the colon of recipient mice. In this assay, epithelial injury is initiated at the distal part of colon by the administration of dextran sulfate sodium, and organoids are infused into the luminal space via the anus. The infused organoids subsequently attach to the injured region and rebuild a donor-derived epithelium. The steps for cell infusion can be completed in 10 min. The assay has been applied successfully to organoids derived from both wild-type and genetically altered epithelial cells from adult colonic and small intestinal epithelium, as well as fetal small intestine. This is a versatile protocol, providing the technical basis for transplantation following alternative colonic injury models. It has been used previously for functional assays to probe cellular potential, and formed the basis for the first in-human clinical trial using colonic organoid transplantation therapy for intractable cases of ulcerative colitis.
肠类器官是一种重要的体外工具，它为肠道干细胞研究提供了新的研究机会。类器官也可以在体内移植，这使它们能够探测干细胞的潜力，并用于疾病建模和再生医学的临床前工具。在这里，我们详细描述了如何将上皮类器官原位移植到受体小鼠的结肠中。在这个实验中，通过给予葡聚糖硫酸钠在远端结肠引发上皮损伤，然后通过肛门将类器官注入腔隙。注入的类器官随后附着在损伤区域，并重建供体衍生的上皮。细胞输注的步骤可以在 10 分钟内完成。该实验已成功应用于源自成年结肠和小肠上皮的野生型和基因改变的上皮细胞以及胎鼠小肠的类器官。这是一个多功能的方案，为替代性结肠损伤模型的移植提供了技术基础。它以前曾被用于功能测定以探测细胞潜力，并为使用结肠类器官移植治疗溃疡性结肠炎的难治性病例进行的首例人体临床试验提供了依据。
