Exposure of lung tissues to cigarette smoke is a major cause of human disease and death worldwide. Unfortunately, adequate model systems that can reliably recapitulate disease biogenesis in vitro, including exposure of the human lung airway to fresh whole cigarette smoke (WCS) under physiological breathing airflow, are lacking. This protocol extension builds upon, and can be used with, our earlier protocol for microfabrication of human organs-on-chips. Here, we describe the engineering, assembly and operation of a microfluidically coupled, multi-compartment platform that bidirectionally ‘breathes’ WCS through microchannels of a human lung small airway microfluidic culture device, mimicking how lung cells may experience smoke in vivo. Several WCS-exposure systems have been developed, but they introduce smoke directly from above the cell cultures, rather than tangentially as naturally occurs in the lung due to lateral airflow. We detail the development of an organ chip–compatible microrespirator and a smoke machine to simulate breathing behavior and smoking topography parameters such as puff time, inter-puff interval and puffs per cigarette. Detailed design files, assembly instructions and control software are provided. This novel platform can be fabricated and assembled in days and can be used repeatedly. Moderate to advanced engineering and programming skills are required to successfully implement this protocol. When coupled with the small airway chip, this protocol can enable prediction of patient-specific biological responses in a matched-comparative manner. We also demonstrate how to adapt the protocol to expose living ciliated airway epithelial cells to smoke generated by electronic cigarettes (e-cigarettes) on-chip.

肺组织暴露于香烟烟雾是全世界人类疾病和死亡的主要原因。不幸的是，缺乏能够在体外可靠地概括疾病生物发生的足够模型系统，包括在生理呼吸气流下将人肺气道暴露于新鲜的全香烟烟雾 (WCS) 中。该协议扩展建立在我们早期用于人体器官芯片微制造的协议之上，并且可以与该协议一起使用。在这里，我们描述了微流体耦合、多隔室平台的工程、组装和操作，该平台通过人肺小气道微流体培养装置的微通道双向“呼吸”WCS，模拟肺细胞如何在体内体验烟雾。已经开发了几种 WCS 暴露系统，但它们直接从细胞培养物上方引入烟雾，而不是由于侧向气流而在肺中自然发生的切向。我们详细介绍了与器官芯片兼容的微型呼吸器和烟雾机的开发，以模拟呼吸行为和吸烟地形参数，例如抽吸时间、抽吸间隔和每支香烟的抽吸次数。提供详细的设计文件、组装说明和控制软件。这种新颖的平台可以在几天内制造和组装，并且可以重复使用。成功实施此协议需要中到高级的工程和编程技能。当与小气道芯片结合使用时，该协议可以以匹配比较的方式预测患者特定的生物反应。