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pp 1-9 | Cite as

Efficient Culture of Intestinal Organoids with Blebbistatin

  • Zhen Qi
  • Ye-Guang Chen
Protocol
Part of the Methods in Molecular Biology book series

Abstract

The intestinal epithelium is one of the most rapidly self-renewing tissues throughout life in mammals. A small population of stem cells at the base of crypt in the epithelium can continually self-renew and give rise to differentiated epithelial cells. The self-renewal and differentiation of intestinal stem cells are under a tight control during homeostasis, and disruption of this balancing regulation leads to intestinal degeneration or tumorigenesis. Accordingly, exploration of the mechanism underlying the regulation of stem cells is essential for the understanding and treatment of intestinal disorders. As traditional methods using mice models are costly and time-consuming, the recently established ex vivo intestinal organoids model provides an ideal tool to investigate the mechanisms regulating the self-renewal and differentiation of intestinal stem cells. The intestinal organoids recapitulate major characteristics in both structure and function of intestinal epithelium in vivo. Here, we describe a new protocol to generate the intestinal organoids from both crypts and single stem cells with a higher efficiency using the small molecule blebbistatin and provide an approach to assess the self-renewal and differentiation of stem cells in intestinal organoids.

Keywords

Intestinal organoids Lgr5+ stem cells Blebbistatin Culture Self-renewal Differentiation 

Notes

Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (31330049,91519310) and the 973 Program (2013CB933700) to YGC.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.The State Key Laboratory of Membrane Biology, Tsinghua-Peking Center for Life Sciences, School of Life SciencesTsinghua UniversityBeijingChina

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