Microfabrication of human organs-on-chips
Microfabrication of human organs-on-chips
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DOI：
10.1038/nprot.2013.137
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发表时间：
2013-11-01
期刊：
影响因子：
14.8
通讯作者：
Ingber, Donald E.
中科院分区：
文献类型：
--
作者：
Huh, Dongeun;Kim, Hyun Jung;Ingber, Donald E.
“器官芯片”是微工程仿生系统，包含由活的人类细胞排列的微流体通道，其复制活器官的关键功能单元，以在体外重建完整的人类器官水平的病理生理学。这些微型设备可用于测试药物和化学品的功效和毒性，并创建人类疾病的体外模型。因此，它们可能代表用于制药、化学和环境应用的常规动物模型的低成本替代品。在这里，我们描述了一个协议的制造，微工程和操作这些微流体器官芯片系统。首先，微工程是用来制造一个多层的微流体装置，其中包含两个平行的弹性体微通道分开的薄多孔柔性膜，沿着与两个全高，中空的真空室在任何一侧，这需要类似的3.5 d完成。为了创建模拟活体人肺的机械活性肺泡-毛细血管界面的“呼吸”肺芯片，在微装置中培养人肺泡上皮细胞和微血管内皮细胞，向侧腔室施加生理流动和循环抽吸以再现有节奏的呼吸运动。我们描述了该协议如何可以很容易地适用于开发其他人体器官芯片，如由人类肠上皮细胞排列的芯片上的肠道，其经历了类似膀胱炎的运动和滴流的流体流动。此外，我们还讨论了可用于分析这些器官芯片设备中的细胞的实验技术。
'Organs-on-chips' are microengineered biomimetic systems containing microfluidic channels lined by living human cells, which replicate key functional units of living organs to reconstitute integrated human organ-level pathophysiology in vitro. These microdevices can be used to test efficacy and toxicity of drugs and chemicals, and to create in vitro models of human disease. Thus, they potentially represent low-cost alternatives to conventional animal models for pharmaceutical, chemical and environmental applications. Here we describe a protocol for the fabrication, microengineering and operation of these microfluidic organ-on-chip systems. First, microengineering is used to fabricate a multilayered microfluidic device that contains two parallel elastomeric microchannels separated by a thin porous flexible membrane, along with two full-height, hollow vacuum chambers on either side; this requires similar to 3.5 d to complete. To create a 'breathing' lung-on-a-chip that mimics the mechanically active alveolar-capillary interface of the living human lung, human alveolar epithelial cells and microvascular endothelial cells are cultured in the microdevice with physiological flow and cyclic suction applied to the side chambers to reproduce rhythmic breathing movements. We describe how this protocol can be easily adapted to develop other human organ chips, such as a gut-on-a-chip lined by human intestinal epithelial cells that experiences peristalsis-like motions and trickling fluid flow. Also, we discuss experimental techniques that can be used to analyze the cells in these organ-on-chip devices.