Co-culturing Human Intestinal Enteroid Monolayers with Innate Immune Cells.
Co-culturing Human Intestinal Enteroid Monolayers with Innate Immune Cells.
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人肠肠样单层细胞与先天免疫细胞共培养。
DOI：
10.1007/978-1-0716-3076-1_16
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Zachos,NicholasC
中科院分区：
文献类型：
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作者：
Staab,JanetF;Lemme-Dumit,JoseM;Latanich,Rachel;Pasetti,MarcelaF;Zachos,NicholasC
肠上皮和免疫细胞之间的协调相互作用是维持适当的屏障功能和粘膜宿主防御肠腔的恶劣外部环境所必需的。作为体内模型的补充，需要一种实用的和可重复的体外模型，该模型采用原代人细胞来确认和促进我们对生理和病理生理条件下粘膜免疫应答的理解。在这里，我们描述了将在可渗透支持物上生长为汇合单层的人肠干细胞衍生的类肠细胞与原代人先天免疫细胞（例如，单核细胞衍生的巨噬细胞和多形核嗜中性粒细胞）。该共培养模型重建了人肠上皮免疫小生境的细胞框架，具有不同的顶侧和基底侧隔室，以分别重建宿主对管腔和粘膜下挑战的反应。肠样免疫共培养物使得能够进行多种结果测量以询问重要的生物过程，例如上皮屏障完整性、干细胞生物学、细胞可塑性、上皮免疫细胞串扰、免疫细胞效应子功能、基因表达的变化（即，转录组学、蛋白质组学、表观遗传学）和宿主-微生物组相互作用。
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.