Prostaglandin E2 supports growth of chicken embryo intestinal organoids in Matrigel matrix

BioTechniques
Malgorzata PierzchalskaPiotr Pierzchalski

Abstract

Investigating intestinal physiology in vitro remains challenging due to the lack of an effective primary enterocyte culture system. Recently developed protocols for growing organoids containing crypts and villus from adult mouse intestinal epithelium in Matrigel present an attractive alternative to the classical techniques. However, these approaches require the use of sophisticated and expensive serum-free medium supplemented with epithelial growth factor (EGF), Wnt agonist (R-spondin 1), and bone morphogenetic protein inhibitor (Noggin) in high concentrations. Here we demonstrate that is possible to use an isolated chicken embryonic intestinal epithelium to create such an organoid culture. Structures formed in Matrigel matrix in the first two days following isolation survive and enlarge during ensuing weeks. They have the appearance of empty spheres and comprise cells expressing cytokeratin (an epithelial cell marker), villin (a marker of enterocytes), and Sox-9 (a transcription factor characteristic of progenitors and stem cells of intestinal crypts). With chicken embryonic tissue as a source of organoids, prostaglandin E2 is as effective as R-spondin 1 and Noggin in promoting sustained growth and survival of epithelial spher...Continue Reading

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Citations

Jan 27, 2018·Cytotechnology·Malgorzata PanekMalgorzata Pierzchalska
Jun 20, 2018·Poultry Science·Narayan C RathJackson O Lay
Jul 20, 2016·Annual Review of Nutrition·Tim Y HouRobert S Chapkin
Jun 7, 2019·Journal of Applied Toxicology : JAT·Justyna AugustyniakFrancesca Caloni
Nov 14, 2019·Animals : an Open Access Journal From MDPI·Haihan ZhangYang Liu
Sep 6, 2018·Journal of Applied Toxicology : JAT·Weijia HanZhongping Duan
Feb 14, 2021·BMC Molecular and Cell Biology·Federico GhiselliEster Grilli
Feb 27, 2021·Veterinary Research·Martin BeaumontAgnès Wiedemann
Mar 12, 2021·Veterinary Research·Soumya K KarHenri Woelders
Feb 12, 2019·Journal of Agricultural and Food Chemistry·Yue-Bang YinYu-Long Yin

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Methods Mentioned

BETA
immunoprecipitation
FCS

Software Mentioned

Statistica
Moticplus
AF6000

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