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Organoids pp 171-181 | Cite as

Murine Colonic Organoid Culture System and Downstream Assay Applications

  • Yang-Yi Fan
  • Laurie A. Davidson
  • Robert S. ChapkinEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1576)

Abstract

Colonic organoids, three-dimensional colonic crypts grown in vitro that show realistic microanatomy, have many potential applications for studying physiology, developmental biology, and pathophysiology of intestinal diseases including inflammatory bowel disease and colorectal cancer. Here, we describe detailed protocols for mouse colonic crypt isolation, organoid culture, and downstream applications. Specific culture strategies including growth factor enriched Matrigel and Wnt and R-spondin conditioned media serve as key factors for enhancing the growth and cost efficiency of colonic organoid cultures.

Keywords:

Niche-independent crypt expansion Stem cells In vitro primary culture 

Notes

Acknowledgement

This work was supported by funding from NIH R35CA197707 and P30ES023512. We would like to thank Drs. Hans Clevers, Nick Barker, and Toshiro Sato for initial guidance in organoid culture technique.

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Copyright information

© Springer Science+Business Media New York 2016

Open Access This chapter is licensed under the terms of the Creative Commons Attribution-NonCommercial 2.5 International License (http://creativecommons.org/licenses/by-nc/2.5/), which permits any noncommercial use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

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Authors and Affiliations

  • Yang-Yi Fan
    • 1
  • Laurie A. Davidson
    • 1
    • 2
  • Robert S. Chapkin
    • 1
    • 2
    Email author
  1. 1.Program in Integrative Nutrition and Complex Diseases, Department of Nutrition & Food ScienceTexas A&M UniversityCollege StationUSA
  2. 2.Center for Translational Environmental Health ResearchTexas A&M UniversityCollege StationUSA

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