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KMID : 1161320220370020136
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Journal of Animal Reproduciton and Biotechnology
2022 Volume.37 No. 2 p.136 ~ p.143
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Effect of Wnt signaling pathway activation on the efficient generation of bovine intestinal organoids
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Park Kang-Won
Yang Hyeon
Wi Ha-Yeon
Ock Sun-A
Lee Poong-Yeon
Hwang In-Sul
Lee Bo-Ram
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Abstract
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Recent progress has been made to establish intestinal organoids for an in vitro model as a potential alternative to an in vivo system in animals. We previously reported a reliable method for the isolation of intestinal crypts from the small intestine and robust three-dimensional (3D) expansion of intestinal organoids (basal-out) in adult bovines. The present study aimed to establish next-generation intestinal organoids for practical applications in disease modeling-based host-pathogen interactions and feed efficiency measurements. In this study, we developed a rapid and convenient method for the efficient generation of intestinal organoids through the modulation of the Wnt signaling pathway and continuous apical-out intestinal organoids. Remarkably, the intestinal epithelium only takes 3-4 days to undergo CHIR (1μM) treatment as a Wnt activator, which is much shorter than that required for spontaneous differentiation (7 days). Subsequently, we successfully established an apical-out bovine intestinal organoid culture system through suspension culture without Matrigel matrix, indicating an apical-out membrane on the surface. Collectively, these results demonstrate the efficient generation and next-generation of bovine intestinal organoids and will facilitate their potential use for various purposes, such as disease modeling, in the field of animal biotechnology.
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KEYWORD
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apical-out, bovine, intestinal organoid, Wnt signaling pathway activation
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