Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), 77 Cheongam-ro, Nam-gu, Pohang, Gyeongbuk 37673, South Korea.
Lab Chip. 2021 Sep 7;21(17):3316-3327. doi: 10.1039/d1lc00404b. Epub 2021 Jul 29.
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.
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再现人类肠道上皮的三维（3D）微结构的多个复制。“基底外侧对流产生多孔插入平台（BASIN）”包含 24 个纳米多孔插入物和一个开放的基底外侧腔室，在基底外侧腔室中施加可控的对流流，使用常规的轨道摇床再现仿生形态发生梯度。通过去除基底外侧培养基中的形态发生抑制剂来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 BASIN 中，我们成功地使用 Caco-2 人肠道上皮再生了 3D 绒毛样肠道微结构，该上皮具有高屏障功能，并且插入物之间的差异最小。使用吸收（绒毛蛋白）和增殖细胞（Ki67）的标志物可视化了 BASIN 中形成的 3D 上皮层的增强的细胞分化和增殖。微工程 3D 上皮层在 BASIN 中的旁分泌转运和外排谱证实了其重现性、鲁棒性和可扩展性，适用于多重生化或药物研究。最后，使用 BASIN 研究了葡聚糖硫酸钠对肠道上皮屏障和形态的影响，验证了其用于研究外部化学物质对肠道上皮的影响和构建漏肠模型的实际适用性。我们设想，BASIN 可以提供一种改进的多重、可扩展和生理肠道上皮模型，便于基础科学和应用科学的研究人员使用。
