Here, we report a multiplex culture system that enables simultaneous recreation of multiple replications of the three-dimensional (3D) microarchitecture of the human intestinal epithelium in vitro. The “basolateral convective flow-generating multi-well insert platform (BASIN)” contains 24 nano-porous inserts and an open basolateral chamber applying controllable convective flow in the basolateral compartment that recreates a biomimetic morphogen gradient using a conventional orbital shaker. The mechanistic approach by which the removal of morphogen inhibitors in the basolateral medium can induce intestinal morphogenesis was applied to manipulate the basolateral convective flow in space and time. In a multiplex BASIN, we successfully regenerated a 3D villi-like intestinal microstructure using the Caco-2 human intestinal epithelium that presents high barrier function with minimal insert-to-insert variations. The enhanced cytodifferentiation and proliferation of the 3D epithelial layers formed in the BASIN were visualized with markers of absorptive (villin) and proliferative cells (Ki67). The paracellular transport and efflux profiles of the microengineered 3D epithelial layers in the BASIN confirmed its reproducibility, robustness, and scalability for multiplex biochemical or pharmaceutical studies. Finally, the BASIN was used to investigate the effects of dextran sodium sulfate on the intestinal epithelial barrier and morphology to validate its practical applicability for investigating the effects of external chemicals on the intestinal epithelium and constructing a leaky-gut model. We envision that the BASIN may provide an improved multiplex, scalable, and physiological intestinal epithelial model that is readily accessible to researchers in both basic and applied sciences.

中文翻译：

---

通过基底外侧对流在多孔插入平台上多重再现人类肠道形态发生

在这里，我们报告了一种多重培养系统，该系统能够在体外同时重现人类肠道上皮的三维 (3D) 微结构的多次复制. “基底外侧对流流动多孔插入平台（BASIN）”包含 24 个纳米多孔插件和一个开放的基底外侧室，在基底外侧隔室中应用可控对流流动，使用传统的轨道振动器重建仿生形态发生梯度。在基底外侧介质中去除形态发生素抑制剂可以诱导肠道形态发生的机械方法被应用于操纵基底外侧对流在空间和时间上的流动。在多重盆地中，我们使用 Caco-2 人肠上皮成功再生了 3D 绒毛状肠道微结构，该上皮具有高屏障功能，插入物与插入物之间的差异最小。在 BASIN 中形成的 3D 上皮层的增强的细胞分化和增殖通过吸收性（绒毛蛋白）和增殖性细胞（Ki67）的标记进行可视化。BASIN 中微工程 3D 上皮层的细胞旁转运和流出曲线证实了其在多重生化或药物研究中的可重复性、稳健性和可扩展性。最后，使用BASIN研究葡聚糖硫酸钠对肠上皮屏障和形态的影响，以验证其在研究外部化学物质对肠上皮的影响和构建漏肠模型方面的实际适用性。我们设想 BASIN 可以提供改进的多重、可扩展、