Background and Aims: There is increasing evidence that study of normal human enteroids duplicates many known aspects of human intestinal physiology. However, this epithelial cell-only model lacks the many non-epithelial intestinal cells present in the gastrointestinal tract and exposure to the mechanical forces to which the intestine is exposed. We tested the hypothesis that physical shear forces produced by luminal and blood flow would provide an intestinal model more closely resembling normal human jejunum. Methods: Jejunal enteroid monolayers were studied in the Emulate, Inc Intestine-Chip under conditions of constant luminal and basolateral flow that was designed to mimic normal intestinal fluid flow, with human umbilical vein endothelial cells (HUVECs) on the basolateral surface and with Wnt3A, R-spondin, Noggin only on the luminal surface. Results: The jejunal enteroids formed monolayers that remained confluent for 6-8 days; began differentiating at least as early as day two post-plating, and demonstrated continuing differentiation over the entire time of the study as shown by quantitative real-time polymerase chain reaction and Western blotting. These results were consistent with continual differentiation, as was shown to occur in mouse villus enterocytes. Compared to differentiated enteroid monolayers grown on Transwell inserts, enteroids exposed to flow were more differentiated but exhibited increased apoptosis and reduced carbohydrate metabolism as shown by proteomic analysis. Conclusions: This study of human jejunal enteroids-on-chip suggests that luminal and basolateral flow produce a model of continual differentiation over time and NaCl absorption that mimics normal intestine and should provide new insights in intestinal physiology.

中文翻译：

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流体剪切应力增强肠芯片中空肠人肠样的分化

背景和目的：越来越多的证据表明，对正常人体肠样的研究重复了人体肠道生理学的许多已知方面。然而，这种只有上皮细胞的模型缺乏胃肠道中存在的许多非上皮肠细胞，也缺乏暴露于肠道所暴露的机械力。我们测试了一个假设，即管腔和血流产生的物理剪切力将提供更类似于正常人空肠的肠道模型。方R-spondin，Noggin 仅在管腔表面。结果：空肠小肠形成单层，并在 6-8 天内保持融合；至少早在接种后第二天就开始分化，并通过实时定量聚合酶链反应和蛋白质印迹显示在整个研究时间内持续分化。这些结果与持续分化一致，正如在小鼠绒毛肠细胞中所显示的那样。与在 Transwell 插入物上生长的分化肠样单层相比，暴露于流动的肠样更加分化，但表现出增加的细胞凋亡和减少的碳水化合物代谢，如蛋白质组学分析所示。结论：