Morphofunctional Characterization of Colonic Organoids from Muc2-/- Mice

Popova, J. V., Medvedeva, S. S., Zybkina, A. V., Kozhevnikova, E. N.

10.64898/2026.10.21.760621 · was preprinted

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Abstract

Inflammatory bowel diseases (IBD) remain a serious challenge in gastroenterology, and disruption of the mucus barrier integrity plays a key role in their pathogenesis. The Muc2 protein is the main structural component of the protective mucus layer in the colon, and mice with a knockout of the Muc2 gene (Muc2-/-) serve as a classic model for studying IBD. In the present work, three-dimensional cultured organoids derived from the colonic crypts of Muc2-/- mice and their wild-type littermates (Muc2/) were used for a comparative analysis of their morphofunctional characteristics. Using morphometric analysis and flow cytometry to identify the Lgr5 stem cell population, we demonstrated that organoids obtained from Muc2-/- animals are characterized by a smaller size, absence of crypt-like structures, increased proliferative activity, and a reduced proportion of stem cells compared to organoids derived from wild-type littermates. The obtained data indicate that Muc2 deficiency and the associated intestinal inflammation affect epithelial architecture and stem cell balance outside the inflammatory environment in vitro. Thus, 3D colonic organoids derived from Muc2 knockout mice represent a promising tool for studying IBD and testing novel therapeutic strategies.

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