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Organoids pp 135-144 | Cite as

The Three-Dimensional Culture of Epithelial Organoids Derived from Embryonic Chicken Intestine

  • Malgorzata PierzchalskaEmail author
  • Malgorzata Panek
  • Malgorzata Czyrnek
  • Maja Grabacka
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1576)

Abstract

The intestinal epithelium isolated from chicken embryos in last 3 days of development can be used to establish the 3D culture of intestinal organoids. When fragments of epithelial tissue released by incubation with EGTA (2.5 mM, 2 h) are embedded in Matrigel matrix on cell culture inserts the formation of empty spheres covered by epithelial cells is observed in first 24 h of culture. The growth and survival of organoids are supported by the addition of R-spondin 1, Noggin, and prostaglandin E2 to the culture medium. The organoids are accompanied by myofibroblasts which become visible in the next 2 days of culture. The intestinal enteroids (free of myofibroblasts) can be obtained from adult chicken intestine.

Keywords:

Intestinal epithelium Three-dimensional cell culture Epithelial organoids Chicken embryo Intestinal myofibroblasts 

Notes

Acknowledgment

The authors acknowledge the financial support of Polish National Science Center (grant no. 2013/09/B/NZ9/00285 to M.P.)

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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

  • Malgorzata Pierzchalska
    • 1
    Email author
  • Malgorzata Panek
    • 1
  • Malgorzata Czyrnek
    • 1
  • Maja Grabacka
    • 1
  1. 1.Department of Food Biotechnology, Faculty of Food TechnologyThe University of Agriculture in KrakówKrakówPoland

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