Abstract

Intestinal organoids are advanced cellular models, which are widely used in mammalian studies to mimic and study in vivo intestinal function and host–pathogen interactions. Growth factors WNT3 and RSPO1 are crucial for the growth of intestinal organoids. Chicken intestinal organoids are currently cultured with mammalian Wnt3a and Rspo1, however, maintaining their longevity has shown to be challenging. Based on the limited homology between mammalian and avian RSPO1, we expect that chicken-derived factors are required for the organoid cultures. Isolated crypts from embryonic tissue of laying hens were growing in the presence of chicken WNT3 and RSPO1, whereas growth in the presence of mammalian Wnt3a and Rspo1 was limited. Moreover, the growth was increased by using Prostaglandin E2 (PGE2) and a Forkhead box O1-inhibitor (FOXO1-inhibitor), allowing to culture these organoids for 15 passages. Furthermore, stem cells maintained their ability to differentiate into goblets, enterocytes and enteroendocrine cells in 2D structures. Overall, we show that chicken intestinal organoids can be cultured for multiple passages using chicken-derived WNT3 and RSPO1, PGE2, and FOXO1-inhibitor.

Details

Title
Chicken-derived RSPO1 and WNT3 contribute to maintaining longevity of chicken intestinal organoid cultures
Author
Oost, Miriam J. 1 ; Ijaz, Adil 2 ; van Haarlem, Daphne A. 2 ; van Summeren, Kitty 3 ; Velkers, Francisca C. 4 ; Kraneveld, Aletta D. 5 ; Venema, Koen 1 ; Jansen, Christine A. 6 ; Pieters, Raymond H. H. 7 ; ten Klooster, Jean Paul 3 

 Maastricht University, Centre for Healthy Eating and Food Innovation, Faculty of Science and Engineering, Venlo, The Netherlands (GRID:grid.5012.6) (ISNI:0000 0001 0481 6099) 
 Utrecht University, Division Infectious Diseases and Immunology, Department Biomolecular Health Sciences, Faculty of Veterinary Medicine, Utrecht, The Netherlands (GRID:grid.5477.1) (ISNI:0000000120346234) 
 University of Applied Sciences Utrecht, Innovative Testing in Life Sciences and Chemistry, Research Centre Healthy and Sustainable Living, Utrecht, The Netherlands (GRID:grid.438049.2) (ISNI:0000 0001 0824 9343) 
 Utrecht University, Division Farm Animal Health, Department Population Health Sciences, Faculty of Veterinary Medicine, Utrecht, The Netherlands (GRID:grid.5477.1) (ISNI:0000000120346234) 
 Utrecht University, Division of Pharmacology, Utrecht Institute for Pharmaceutical Sciences, Faculty of Science, Utrecht, The Netherlands (GRID:grid.5477.1) (ISNI:0000000120346234) 
 Utrecht University, Division Infectious Diseases and Immunology, Department Biomolecular Health Sciences, Faculty of Veterinary Medicine, Utrecht, The Netherlands (GRID:grid.5477.1) (ISNI:0000000120346234); Wageningen University & Research, Department of Animal Sciences, Cell Biology and Immunology Group, Wageningen, The Netherlands (GRID:grid.4818.5) (ISNI:0000 0001 0791 5666) 
 University of Applied Sciences Utrecht, Innovative Testing in Life Sciences and Chemistry, Research Centre Healthy and Sustainable Living, Utrecht, The Netherlands (GRID:grid.438049.2) (ISNI:0000 0001 0824 9343); Utrecht University, Department of Population Health Sciences, Institute for Risk Assessment Sciences, Utrecht, The Netherlands (GRID:grid.5477.1) (ISNI:0000000120346234) 
Publication year
2022
Publication date
2022
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2679469727
Copyright
© The Author(s) 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.