Background: The development and maturation of intestinal stem cells (ISCs) into absorptive and secretory epithelial cells is controlled by multiple types of stromal cells, including immune cells. Mice that lack innate immune cells have reduced intestinal surface area, a result mimicked in knockout mice lacking specific cytokines. These data suggest that resident immune cell factors help regulate intestinal epithelial proliferation during homeostasis. In contrast, exposure of the intestinal epithelium to high levels of inflammation, such as in Inflammatory Bowel Diseases (IBD), can result in epithelial dysfunction and/or cell death. While much of our understanding of IBD pathogenesis comes from mouse models, human intestinal organoids generated from IBD patients recapitulate transcriptional changes characteristic of disease suggesting that disease-specific defects are imprinted in intestinal stem cells. Our data support the hypothesis that cytokines differentially affect ISC proliferation, lineage differentiation, barrier function, and wound repair in human intestinal organoids from healthy donors compared to those of IBD patients. Aims: The role of specific cytokines and their effects under normal or pro-inflammatory conditions has not been established in human models. We aim to identify and characterize the role of cytokines from gut resident innate immune cells on intestinal stem cell homeostasis and epithelial barrier function as well as their contributions to epithelial regeneration patients with active and inactive IBD. Methods: Human intestinal organoids were derived from biopsies obtained from healthy donors or from inflamed and non-inflamed areas of patients with Ulcerative colitis (UC) or ileal Crohn’s disease (CD). Cultures were placed on Transwells to form 2D monolayers and exposed to recombinant human cytokines for various time points (up to 3 months). Cytokine effects on organoid growth and wound repair were observed by time lapse microscopy and transcription changes were examined by qPCR. Results: Organoids exposed to IL-6 or TNF-α showed changes in LGR5 expression. These changes were not consistent between regions. Colonoids had increased OLMF4. Additionally, IL-6 inversely affected lineage differentiation by increasing enteroendocrine cells in enteroids and Paneth cells in colonoids. Under basal conditions, CD and UC organoids from areas of active inflammation had reduced plating efficiency and long-term survival compared to organoids from non-inflamed areas from the same patient. Active UC colonoids had abnormal barrier function and repair following wound assay in monolayers compared to healthy controls and/or inactive UC colonoids. On the other hand, active CD enteroids had fewer actively dividing ISCs and Paneth cells with increased numbers of enteroendocrine and Goblet cells compared to inactive CD or healthy control enteroids. Conclusions: Human intestinal organoids derived from active areas of inflammation in IBD patients exhibit a) defects in epithelial survival and function and b) differential effects of cytokines on ISC markers, culture growth and survival, as well as response to injury. This abstract was presented at the American Physiology Summit 2026 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.
Sanchez et al. (Fri,) studied this question.
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