Abstract
The intestinal epithelium is the most rapidly renewed tissue in adult mammals, and its renewal is strictly controlled by intestinal stem cells. Extensive studies using genetic models of intestinal epithelium have revealed the mechanisms underlying the self-renewal of intestinal stem cells. Exploiting this knowledge, we developed a novel 3D culture system that enables the outgrowth of intestinal Lgr5+ stem cells derived from mouse and human tissues into ever-expanding crypt–villus mini-guts, known as intestinal epithelial organoids. These organoids are maintained by the self-renewal of stem cells and give rise to all differentiated cell types of the intestinal epithelium. Once established, organoids can be cryopreserved and thawed when needed. This culture system has been widely used for studying stem cell behavior and gene function and for disease modeling.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Barker N, van Es JH, Kuipers J et al (2007) Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449:1003–1007
Sato T, Vries RG, Snippert HJ et al (2009) Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459:262–265
Sato T, Stange DE, Ferrante M et al (2011) Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett’s epithelium. Gastroenterology 141:1762–1772
Watanabe K, Ueno M, Kamiya D et al (2007) A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nat Biotechnol 25:681–686
Sato T, Clevers H (2013) Growing self-organizing mini-guts from a single intestinal stem cell: mechanism and applications. Science 340:1190–1194
Ootani A, Li X, Sangiorgi E et al (2009) Sustained in vitro intestinal epithelial culture within a Wnt-dependent stem cell niche. Nat Med 15:701–706
Farin HF, Van Es JH, Clevers H (2012) Redundant sources of Wnt regulate intestinal stem cells and promote formation of Paneth cells. Gastroenterology 143:1518–1529
Willert K, Brown JD, Danenberg E et al (2003) Wnt proteins are lipid-modified and can act as stem cell growth factors. Nature 423:448–452
Korinek V, Barker N, Morin PJ et al (1997) Constitutive transcriptional activation by a beta-catenin-Tcf complex in APC−/− colon carcinoma. Science 275:1784–1787
Acknowledgments
We thank the past and present members of the Sato laboratory for their continued support. S.S. was supported by the Japan Society for the Promotion of Science Research Fellowships for Young Scientists. This work was supported in part by JSPS KAKENHI Grant Number 15J00981.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Sugimoto, S., Sato, T. (2017). Establishment of 3D Intestinal Organoid Cultures from Intestinal Stem Cells. In: Koledova, Z. (eds) 3D Cell Culture. Methods in Molecular Biology, vol 1612. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7021-6_7
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7021-6_7
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7019-3
Online ISBN: 978-1-4939-7021-6
eBook Packages: Springer Protocols