Transplantation of intestinal organoids into a mouse model of colitis
Transplantation of intestinal organoids into a mouse model of colitis
复制标题
DOI：
10.1038/s41596-021-00658-3
复制
发表时间：
2022-02-02
期刊：
影响因子：
14.8
通讯作者：
Yui, Shiro
中科院分区：
文献类型：
--
作者：
Watanabe, Satoshi;Kobayashi, Sakurako;Yui, Shiro
肠道类器官是体外研究的基本工具，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，并用于疾病建模和作为再生医学的临床前工具。在这里，我们详细描述了如何原位移植上皮类器官到受体小鼠的结肠。在该测定中，通过施用葡聚糖硫酸钠在结肠的远端部分处引发上皮损伤，并且将类器官经由肛门输注到管腔空间中。输注的类器官随后附着在受损区域并重建供体来源的上皮。细胞输注的步骤可以在10分钟内完成。该测定已成功应用于来自成人结肠和小肠上皮以及胎儿小肠的野生型和遗传改变的上皮细胞的类器官。这是一个通用的协议，提供了技术基础移植后，替代结肠损伤模型。它以前已用于功能测定以探测细胞潜能，并形成了第一个在-结肠类器官移植治疗溃疡性结肠炎顽固性病例的人体临床试验：研究胚胎或成人状态的类器官以及小肠和结肠通过移植到动物体内的上皮细胞的方法，在动物中，给予葡聚糖后结肠已受损硫酸钠
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.An approach to studying epithelia derived from organoids of either fetal or adult state and from both the small intestine and colon via transplantation into animals in which the colon has been damaged following administration of dextran sulfate sodium.