Department of Organoid Medicine, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
Department of Gastroenterology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.
Methods Mol Biol. 2020;2171:303-320. doi: 10.1007/978-1-0716-0747-3_21.
Intestinal stem cells continuously self-renew throughout life to maintain gut homeostasis. With the advent of the organoid culture system, we are now able to indefinitely expand healthy and diseased tissue-derived human intestinal stem cells in vitro and use them for various applications. Nonetheless, investigating the behavior of human intestinal stem cells in vivo still remains challenging. We recently developed an orthotopic xenotransplantation system that realizes in vivo reconstruction of human intestinal epithelial tissue with preserved stem cell hierarchy by engrafting human normal colon organoids onto the mouse colon surface. We also introduced new growth factors, namely, insulin-like growth factor-1 (IGF-1) and fibroblast growth factor-2 (FGF-2), into the culture condition for human intestinal organoids that significantly increase scalability and transfectability of the organoids. By integrating these recent advances, we organized a tissue-oriented platform encompassing derivation of patient-derived intestinal organoids and their orthotopic xenotransplantation. The research platform based on orthotopic xenotransplantation of human intestinal organoids provides a powerful tool for studying human intestinal stem cell biology in native tissue-relevant contexts as well as for establishing novel disease modeling systems.
肠干细胞在整个生命过程中持续自我更新，以维持肠道内稳态。随着类器官培养系统的出现，我们现在能够在体外无限扩增健康和疾病组织来源的人类肠干细胞，并将其用于各种应用。尽管如此，研究人类肠干细胞的体内行为仍然具有挑战性。我们最近开发了一种原位异种移植系统，通过将人正常结肠类器官移植到小鼠结肠表面，实现了保留干细胞层次结构的人肠道上皮组织的体内重建。我们还在人肠道类器官的培养条件中引入了新的生长因子，即胰岛素样生长因子-1（IGF-1）和成纤维细胞生长因子-2（FGF-2），这显著提高了类器官的可扩展性和转染效率。通过整合这些最新进展，我们组织了一个面向组织的平台，包括患者来源的肠道类器官的衍生和它们的原位异种移植。基于人肠道类器官原位异种移植的研究平台为研究人类肠道干细胞在天然组织相关环境中的生物学特性以及建立新的疾病建模系统提供了有力工具。
