Chicken-derived RSPO1 and WNT3 contribute to maintaining longevity of chicken intestinal organoid cultures
Oost, Miriam J; Ijaz, Adil; van Haarlem, Daphne A; van Summeren, Kitty; Velkers, Francisca C; Kraneveld, Aletta D; Venema, Koen; Jansen, Christine A; Pieters, Raymond H H; Ten Klooster, Jean Paul
(2022) Scientific Reports, volume 12, issue 1, pp. 1 - 12
(Article)
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
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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 (PGE 2) 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, PGE 2, and FOXO1-inhibitor.
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Keywords: Animals, Chickens, Dinoprostone/metabolism, Female, Intestinal Mucosa, Intestines, Mammals, Organoids/metabolism, Stem Cells, General
ISSN: 2045-2322
Publisher: NLM (Medline)
Note: Funding Information: This research was performed in the public–private partnership “CarboBiotics” project number LWCC.2017.010 coordinated by the Carbohydrate Competence Center (CCC, https://www.cccresearch.nl ). CarboBiotics was jointly financed by participating industrial partners Cooperatie Avebe U.A., FrieslandCampina Nederland B.V., Nutrition Sciences N.V., and allowances of The Dutch Research Council (NWO). Furthermore, the study was also partly funded by the Centre for Healthy Eating & Food Innovation (HEFI) of Maastricht University—Campus Venlo. This research has been made possible with the support of the Dutch province of Limburg with a grant to HEFI. A.I. is the recipient of an international PhD fellowship from the Punjab Educational Endowment Fund, Punjab, Pakistan. Funding Information: We wish to thank Judith Hendriks for her help with the preparation of the intestinal cryosections. Moreover, we would like to thank the animal caretakers of the department of Population Health Sciences, division Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, for their help regarding the chicken eggs. Fluorescent images have been acquired at the Center of Cellular Imaging, Faculty of Veterinary Medicine, Utrecht University. We also would like to thank our undergraduate students Sira Gevers and Karen van den Anker for their assistance in cloning WNT3 and RSPO1 into the E4-plasmid. Publisher Copyright: © 2022, The Author(s).
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