Intestinal Crypt Organoid: Isolation of Intestinal Stem Cells, In Vitro Culture, and Optical Observation.
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.
Department of Surgery, Far Eastern Memorial Hospital, Pan-Chiao, New Taipei, Taiwan.|Department of Chemical Engineering and Materials Science, Yuan Ze University, Chung-Li, Taoyuan, Taiwan.
Yun
Y
Chen
Department of Biomedical Engineering, National Yang-Ming University, Taipei, Taiwan.
Chuan
C
Li
Department of Surgery, Far Eastern Memorial Hospital, Pan-Chiao, New Taipei, Taiwan. yahuitsi@gmail.com.|Department of Chemical Engineering and Materials Science, Yuan Ze University, Chung-Li, Taoyuan, Taiwan. yahuitsi@gmail.com.
Ya-Hui
YH
Tsai
Department of Surgery, National Taiwan University Hospital, National Taiwan University College of Medicine, Taipei, Taiwan. tsh5110@ntu.edu.tw.
Sheng-Hong
SH
Tseng
D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D006801:Humans; D066298:In Vitro Techniques / Q000379:methods*; D007422:Intestines / Q000166:cytology* / Q000648:ultrastructure; D018613:Microscopy, Confocal / Q000379:methods*; D009940:Organoids / Q000166:cytology* / Q000648:ultrastructure; D013234:Stem Cells / Q000166:cytology* / Q000648:ultrastructure
Confocal observation; Crypt organoid; Intestinal stem cell (ISC); Isolation; Three-dimensional imaging