Department of Biomedical Engineering, Rochester Institute of Technology.
Department of Biomedical Engineering, University of Rochester.
J Vis Exp. 2024 Feb 16(204). doi: 10.3791/66090.
Microphysiological systems are miniaturized cell culture platforms used to mimic the structure and function of human tissues in a laboratory setting. However, these platforms have not gained widespread adoption in bioscience laboratories where open-well, membrane-based approaches serve as the gold standard for mimicking tissue barriers, despite lacking fluid flow capabilities. This issue can be primarily attributed to the incompatibility of existing microphysiological systems with standard protocols and tools developed for open-well systems. Here, we present a protocol for creating a reconfigurable membrane-based platform with an open-well structure, flow enhancement capability, and compatibility with conventional protocols. This system utilizes a magnetic assembly approach that enables reversible switching between open-well and microfluidic modes. With this approach, users have the flexibility to begin an experiment in the open-well format using standard protocols and add or remove flow capabilities as needed. To demonstrate the practical usage of this system and its compatibility with standard techniques, an endothelial cell monolayer was established in an open-well format. The system was reconfigured to introduce fluid flow and then switched to the open-well format to conduct immunostaining and RNA extraction. Due to its compatibility with conventional open-well protocols and flow enhancement capability, this reconfigurable design is expected to be adopted by both engineering and bioscience laboratories.
微生理系统是用于在实验室中模拟人体组织结构和功能的小型化细胞培养平台。然而,这些平台并未在生物科学实验室中得到广泛采用,尽管开放孔、基于膜的方法缺乏流体流动能力,但它们仍是模拟组织屏障的金标准。这个问题主要归因于现有的微生理系统与为开放孔系统开发的标准协议和工具不兼容。在这里,我们提出了一种创建具有开放孔结构、增强流动能力和与传统协议兼容性的可重构基于膜的平台的方案。该系统利用磁组装方法实现了在开放孔和微流控模式之间的可逆切换。通过这种方法,用户可以使用标准协议在开放孔格式下开始实验,并根据需要添加或移除流动能力。为了展示该系统的实际用途及其与标准技术的兼容性,我们在开放孔格式下建立了内皮细胞单层。然后对系统进行重新配置以引入流体流动,然后切换回开放孔格式以进行免疫染色和 RNA 提取。由于其与传统开放孔协议的兼容性和增强流动能力,这种可重构设计预计将被工程和生物科学实验室采用。