Novak Richard, Didier Meredyth, Calamari Elizabeth, Ng Carlos F, Choe Youngjae, Clauson Susan L, Nestor Bret A, Puerta Jefferson, Fleming Rachel, Firoozinezhad Sasan J, Ingber Donald E
Wyss Institute for Biologically Inspired Engineering, Harvard University;
Wyss Institute for Biologically Inspired Engineering, Harvard University; Apple, Inc.
J Vis Exp. 2018 Oct 20(140):58151. doi: 10.3791/58151.
A significant number of lead compounds fail in the pharmaceutical pipeline because animal studies often fail to predict clinical responses in human patients. Human Organ-on-a-Chip (Organ Chip) microfluidic cell culture devices, which provide an experimental in vitro platform to assess efficacy, toxicity, and pharmacokinetic (PK) profiles in humans, may be better predictors of therapeutic efficacy and safety in the clinic compared to animal studies. These devices may be used to model the function of virtually any organ type and can be fluidically linked through common endothelium-lined microchannels to perform in vitro studies on human organ-level and whole body-level physiology without having to conduct experiments on people. These Organ Chips consist of two perfused microfluidic channels separated by a permeable elastomeric membrane with organ-specific parenchymal cells on one side and microvascular endothelium on the other, which can be cyclically stretched to provide organ-specific mechanical cues (e.g., breathing motions in lung). This protocol details the fabrication of flexible, dual channel, Organ Chips through casting of parts using 3D printed molds, enabling combination of multiple casting and post-processing steps. Porous poly (dimethyl siloxane) (PDMS) membranes are cast with micrometer sized through-holes using silicon pillar arrays under compression. Fabrication and assembly of Organ Chips involves equipment and steps that can be implemented outside of a traditional cleanroom. This protocol provides researchers with access to Organ Chip technology for in vitro organ- and body-level studies in drug discovery, safety and efficacy testing, as well as mechanistic studies of fundamental biological processes.
相当数量的先导化合物在制药流程中失败，因为动物研究往往无法预测人类患者的临床反应。人体芯片器官（器官芯片）微流控细胞培养装置为评估人体的疗效、毒性和药代动力学（PK）概况提供了一个体外实验平台，与动物研究相比，它可能是临床治疗效果和安全性的更好预测指标。这些装置可用于模拟几乎任何器官类型的功能，并可通过常见的内皮衬里微通道进行流体连接，以在人体器官水平和全身水平生理学上进行体外研究，而无需在人体上进行实验。这些器官芯片由两个灌注微流控通道组成，中间隔着一层可渗透的弹性膜，一侧是器官特异性实质细胞，另一侧是微血管内皮细胞，可循环拉伸以提供器官特异性机械信号（例如，肺部的呼吸运动）。本方案详细介绍了通过使用3D打印模具铸造零件来制造柔性双通道器官芯片的方法，实现了多个铸造和后处理步骤的组合。多孔聚二甲基硅氧烷（PDMS）膜在压缩状态下使用硅柱阵列铸造出微米尺寸的通孔。器官芯片的制造和组装涉及的设备和步骤可以在传统洁净室之外实施。本方案为研究人员提供了使用器官芯片技术进行药物发现、安全性和有效性测试以及基本生物学过程机制研究中的体外器官和身体水平研究的途径。
