Protocol

Part of the series Methods in Molecular Biology pp 1-14

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Intestinal Crypt Organoid: Isolation of Intestinal Stem Cells, In Vitro Culture, and Optical Observation

  • Yun ChenAffiliated withDepartment of Surgery, Far Eastern Memorial HospitalDepartment of Chemical Engineering and Materials Science, Yuan Ze University
  • , Chuan LiAffiliated withDepartment of Biomedical Engineering, National Yang-Ming University
  • , Ya-Hui TsaiAffiliated withDepartment of Surgery, Far Eastern Memorial HospitalDepartment of Chemical Engineering and Materials Science, Yuan Ze University Email author 
  • , Sheng-Hong TsengAffiliated withDepartment of Surgery, National Taiwan University Hospital, National Taiwan University College of Medicine Email author 

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Abstract

The isolation and culture of intestinal stem cells (ISCs) was first demonstrated in the very recent decade with the identification of ISC marker Lgr5. The growth of ISCs into crypt organoids provides an in vitro model for studying the mucosal physiology, intestinal cancer tumorigenesis, and intestinal regeneration. Here, we describe two different isolation protocols and demonstrate a fixation method that aids in the confocal observation of the organoids.

Keywords:

Intestinal stem cell (ISC) Crypt organoid Isolation Confocal observation Three-dimensional imaging