Rae Brady, Vasse Gwenda F, Mosayebi Jalal, Berge Maarten van den, Pouwels Simon D, Heijink Irene H
Department of Pathology & Medical Biology, University Medical Center Groningen, University of Groningen, 9713 GZ Groningen, The Netherlands.
University Medical Center Groningen, GRIAC Research Institute, University of Groningen, 9713 GZ Groningen, The Netherlands.
Bioengineering (Basel). 2025 Feb 13;12(2):182. doi: 10.3390/bioengineering12020182.
On-chip microfluidics are advanced in vitro models that simulate lung tissue's native 3D environment more closely than static 2D models to investigate the complex lung architecture and multifactorial processes that lead to pulmonary disease. Current microfluidic systems can be restrictive in the quantities of biological sample that can be retrieved from a single micro-channel, such as RNA, protein, and supernatant. Here, we describe a newly developed large-scale airway-on-chip model that employs a surface area for a cell culture wider than that in currently available systems. This enables the collection of samples comparable in volume to traditional cell culture systems, making the device applicable to any workflow utilizing these static systems (RNA isolation, ELISA, etc.). With our construction method, this larger culture area allows for easier handling, the potential for a wide range of exposures, as well as the collection of low-quantity samples (e.g., volatiles or mitochondrial RNA). The model consists of two large polydimethylsiloxane (PDMS) cell culture chambers under an independent flow of medium or air, separated by a semi-permeable polyethylene (PET) cell culture membrane (23 μm thick, 0.4 μm pore size). Each chamber carries a 5 × 18 mm, 90 mm (92 mm with tapered chamber inlets) surface area that can contain up to 1-2 × 10 adherent structural lung cells and can be utilized for close contact co-culture studies of different lung cell types, including airway epithelial cells, fibroblasts, smooth muscle cells, and endothelial cells. The parallel bi-chambered design of the chip allows for epithelial cells to be cultured at the air-liquid interface (ALI) and differentiation into a dense, multi-layered, pseudostratified epithelium under biological flow rates. This millifluidic airway-on-chip advances the field by providing a readily reproducible, easily adjustable, and cost-effective large-scale fluidic 3D airway cell culture platform.
芯片上的微流控技术是先进的体外模型，相较于静态二维模型，它能更紧密地模拟肺组织的天然三维环境，以研究导致肺部疾病的复杂肺结构和多因素过程。当前的微流控系统在从单个微通道中获取的生物样本数量方面可能存在限制，例如RNA、蛋白质和上清液。在此，我们描述了一种新开发的大规模芯片上气道模型，该模型采用了比现有系统更宽的细胞培养表面积。这使得能够收集与传统细胞培养系统体积相当的样本，从而使该设备适用于任何利用这些静态系统的工作流程（RNA分离、酶联免疫吸附测定等）。通过我们的构建方法，这个更大的培养区域便于操作，具有进行广泛暴露的潜力，还能收集少量样本（例如挥发物或线粒体RNA）。该模型由两个大型聚二甲基硅氧烷（PDMS）细胞培养室组成，在独立的培养基或空气流作用下，由半透性聚乙烯（PET）细胞培养膜（厚23μm，孔径0.4μm）分隔。每个培养室的表面积为5×18mm、90mm（带锥形腔室入口时为92mm），可容纳多达1 - 2×10个贴壁的肺结构细胞，可用于不同肺细胞类型的紧密接触共培养研究，包括气道上皮细胞、成纤维细胞、平滑肌细胞和内皮细胞。芯片的平行双腔设计允许上皮细胞在气液界面（ALI）培养，并在生物流速下分化为致密的多层假复层上皮。这种微流控芯片上气道通过提供一个易于重现、易于调节且经济高效的大规模流体三维气道细胞培养平台，推动了该领域的发展。
