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Biol Open. 2017 May 15;6(5):698-705. doi: 10.1242/bio.021717.

WRN conditioned media is sufficient for .

Biology open

Robin H Powell, Michael S Behnke

Affiliations

  1. Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USA.
  2. Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USA [email protected].

PMID: 28347989 PMCID: PMC5450310 DOI: 10.1242/bio.021717

Abstract

Recent years have seen significant developments in the ability to continuously propagate organoids derived from intestinal crypts. These advancements have been applied to mouse and human samples providing models for gastrointestinal tissue development and disease. We adapt these methods for the propagation of intestinal organoids (enteroids) from various large farm and small companion (LF/SC) animals, including cat, dog, cow, horse, pig, sheep and chicken. We show that LF/SC enteroids propagate and expand in L-WRN conditioned media containing signaling factors Wnt3a, R-spondin-3, and Noggin (WRN). Multiple successful isolations were achieved for each species, and the growth of LF/SC enteroids was maintained to high passage number. LF/SC enteroids expressed crypt stem cell marker LGR5 and low levels of mesenchymal marker VIM. Labeling with EdU also showed distinct regions of cell proliferation within the enteroids marking crypt-like regions. The ability to grow and maintain LF/SC enteroid cell lines provides additional models for the study of gastrointestinal developmental biology as well as platforms for the study of host-pathogen interactions between intestinal cells and zoonotic enteric pathogens of medical importance.

© 2017. Published by The Company of Biologists Ltd.

Keywords: Companion animal; Conditioned media; Crypt; Enteroid; Farm animal; Intestine; Organoid

Conflict of interest statement

Competing interestsThe authors declare no competing or financial interests.

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