Enterotoxins increase intestinal fluid secretion through modulation of ion channels as well as activation of the enteric nervous and immune systems. Colonic organoids, also known as colonoids, are functionally and phenotypically similar to in vivo colonic epithelium and have been used to study intestinal ion transport and subsequent water flux in physiology and disease models. In conventional cultures, organoids exist as spheroids embedded within a hydrogel patty of extracellular matrix, and they form at multiple depths, impairing efficient imaging necessary to capture data from statistically relevant sample sizes. To overcome these limitations, an analytical platform with colonic organoids localized to the planar surface of a hydrogel layer was developed. The arrays of densely packed colonoids (140 μm average diameter, 4 colonoids/mm²) were generated in a 96-well plate, enabling assay of the response of hundreds of organoids so that organoid subpopulations with distinct behaviors were identifiable. Organoid cell types, monolayer polarity, and growth were similar to those embedded in hydrogel. An automated imaging and analysis platform efficiently tracked over time swelling due to forskolin and fluid movement across the cell monolayer stimulated by cholera toxin. The platform was used to screen compounds associated with the enteric nervous and immune systems for their effect on fluid movement across epithelial cells. Prostaglandin E2 promoted increased water flux in a subset of organoids that resulted in organoid swelling, confirming a role for this inflammatory mediator in diarrheal conditions but also illustrating organoid differences in response to an identical stimulus. By allowing sampling of a large number of organoids, the arrayed organoid platform permits identification of organoid subpopulations intermixed within a larger group of nonresponding organoids. This technique will enable automated, large-scale screening of the impact of drugs, toxins, and other compounds on colonic physiology.

中文翻译：

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用于筛选与肠毒素有关的促分泌素的阵列结肠类生物体的开发。

肠毒素通过调节离子通道以及激活肠神经和免疫系统来增加肠液分泌。结肠类器官，也称为结肠类，在功能和表型上均与体内结肠上皮相似，并已被用于研究肠道离子运输以及随后的生理和疾病模型中的水通量。在常规培养中，类器官以球状体的形式存在于细胞外基质的水凝胶小块中，并且它们形成于多个深度，从而损害了从统计上相关的样本量中捕获数据所必需的有效成像。为了克服这些局限性，开发了具有结肠类器官位于水凝胶层平面表面的分析平台。密集堆积的类固醇阵列（平均直径140μm，4个类固醇/ mm^2个）是在96孔板中产生的，能够分析数百种类器官的反应，因此可以识别具有不同行为的类器官亚群。器官细胞的类型，单层极性和生长与嵌入水凝胶的细胞相似。自动化的成像和分析平台可有效追踪随时间推移而产生的肿胀，这些肿胀是由于霍乱毒素刺激的毛喉素和液体跨细胞单层运动所致。该平台用于筛选与肠神经和免疫系统相关的化合物，以了解它们对跨上皮细胞的液体运动的影响。前列腺素E2促进了类器官的一部分中的水通量的增​​加，导致类器官肿胀，这证实了这种炎性介质在腹泻条件下的作用，但也说明了类器官对相同刺激的差异。通过允许采样大量的类器官，阵列状的类器官平台可以识别混合在一大堆无响应类器官中的类器官亚群。该技术将能够自动，大规模地筛选药物，毒素和其他化合物对结肠生理的影响。