Department of Chemical and Biological Engineering, Montana State University, Bozeman, MT, USA.
Lab Chip. 2019 Oct 9;19(20):3552-3562. doi: 10.1039/c9lc00653b.
Human intestinal organoids (HIOs) are millimeter-scale models of the human intestinal epithelium and hold tremendous potential for advancing fundamental and applied biomedical research. HIOs resemble the native gut in that they consist of a fluid-filled lumen surrounded by a polarized epithelium and associated mesenchyme; however, their topologically-closed, spherical shape prevents flow through the interior luminal space, making the system less physiological and leading to the buildup of cellular and metabolic waste. These factors ultimately limit experimentation inside the HIOs. Here, we present a millifluidic device called the gut organoid flow chip (GOFlowChip), which we use to "port" HIOs and establish steady-state liquid flow through the lumen for multiple days. This long-term flow is enabled by the use of laser-cut silicone gaskets, which allow liquid in the device to be slightly pressurized, suppressing bubble formation. To demonstrate the utility of the device, we establish separate luminal and extraluminal flow and use luminal flow to remove accumulated waste. This represents the first demonstration of established liquid flow through the luminal space of a gastrointestinal organoid over physiologically relevant time scales. Flow cytometry results reveal that HIO cell viability is unaffected by long-term porting and luminal flow. We expect the real-time, long-term control over luminal and extraluminal contents provided by the GOFlowChip will enable a wide variety of studies including intestinal secretion, absorption, transport, and co-culture with intestinal microorganisms.
人类肠道类器官（HIOs）是毫米级的人类肠道上皮模型，在推进基础和应用生物医学研究方面具有巨大潜力。HIOs 类似于天然肠道，由充满液体的腔室组成，周围是极化的上皮组织和相关的间充质；然而，它们拓扑上封闭的球形形状阻止了内部腔室空间的流动，使系统不太生理，并导致细胞和代谢废物的积累。这些因素最终限制了 HIO 内部的实验。在这里，我们提出了一种叫做肠道类器官流动芯片（GOFlowChip）的毫升级流体装置，我们使用该装置来“移植” HIO 并在腔室内建立稳定的液体流动，持续多日。这种长期流动是通过使用激光切割硅酮垫圈实现的，垫圈允许设备中的液体略微加压，抑制气泡形成。为了演示该设备的实用性，我们建立了单独的腔室和腔外流动，并使用腔室流动来清除积累的废物。这代表了在生理相关时间范围内首次在胃肠道类器官的腔室内建立稳定液体流动的演示。流式细胞术结果表明，长期移植和腔室流动对 HIO 细胞活力没有影响。我们预计，GOFlowChip 提供的实时、长期腔室和腔外内容物控制将能够实现各种研究，包括肠道分泌、吸收、运输以及与肠道微生物的共培养。
