Organoid derivation and orthotopic xenotransplantation for studying human intestinal stem cell dynamics

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages303-320
Number of pages18
DOIs
Publication statusPublished - 2020

Publication series

NameMethods in Molecular Biology
Volume2171
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Colon
  • Crypt isolation
  • Electroporation
  • Endoscopy
  • Human
  • Intestinal stem cells
  • LGR5
  • Mini-gut
  • Organoid culture
  • Orthotopic transplantation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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