Abstract
The coordinated interaction between the intestinal epithelium and immune cells is required to maintain proper barrier function and mucosal host defenses to the harsh external environment of the gut lumen. Complementary to in vivo models, there is a need for practical and reproducible in vitro models that employ primary human cells to confirm and advance our understanding of mucosal immune responses under physiologic and pathophysiologic conditions. Here we describe the methods to co-culture human intestinal stem cell-derived enteroids grown as confluent monolayers on permeable supports with primary human innate immune cells (e.g., monocyte-derived macrophages and polymorphonuclear neutrophils). This co-culture model reconstructs the cellular framework of the human intestinal epithelial-immune niche with distinct apical and basolateral compartments to recreate host responses to luminal and submucosal challenges, respectively. Enteroid-immune co-cultures enable multiple outcome measures to interrogate important biological processes such as epithelial barrier integrity, stem cell biology, cellular plasticity, epithelial-immune cells crosstalk, immune cell effector functions, changes in gene expression (i.e., transcriptomic, proteomic, epigenetic), and host-microbiome interactions.
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Acknowledgments
The protocols developed for co-culture studies were funded by the National Institute of Allergy and Infectious Diseases (P01-AI125181 to MP and NZ). The authors also wish to acknowledge the Integrated Physiology and Imaging Cores of the Hopkins Conte Digestive Disease Basic and Translational Research Core Center (DK-089502 to NZ) for resources used to develop the human immune-enteroid co-culture models.
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Staab, J.F., Lemme-Dumit, J.M., Latanich, R., Pasetti, M.F., Zachos, N.C. (2023). Co-culturing Human Intestinal Enteroid Monolayers with Innate Immune Cells. In: Ordóñez-Morán, P. (eds) Intestinal Differentiated Cells. Methods in Molecular Biology, vol 2650. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3076-1_16
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DOI: https://doi.org/10.1007/978-1-0716-3076-1_16
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