Graduate School of Engineering, Tohoku University, Sendai 980-8579, Japan.
Frontier Research Institute for Interdisciplinary Sciences, Tohoku University, Sendai 980-0845, Japan.
Anal Chem. 2023 Dec 12;95(49):18158-18165. doi: 10.1021/acs.analchem.3c03684. Epub 2023 Nov 28.
Vasculature-on-a-chip is a microfluidic cell culture device used for modeling vascular functions by culturing endothelial cells. Porous membranes are widely used to create cell culture environments. However, in situ real-time measurements of cellular metabolites in microchannels are challenging. In this study, a novel microfluidic device with a porous membrane electrode was developed for the in situ monitoring of nitric oxide (NO) released by endothelial cells in real time. In this system, a porous Au membrane electrode was placed directly beneath the cells for in situ and real-time measurements of NO, a biomarker of endothelial cells. First, the device was electrochemically characterized to construct a calibration plot for NO. Next, NO released by human umbilical vein endothelial cells under l-arginine stimulation was successfully quantified. Furthermore, the changes in NO release with culture time (in days) using the same sample were successfully recorded by exploiting minimally invasive measurements. This is the first report on the combination of a microfluidic device and porous membrane electrode for the electrochemical analysis of endothelial cells. This device will contribute to the development of organ-on-a-chip technology for real-time in situ cell analyses.
血管芯片是一种微流控细胞培养装置，用于通过培养内皮细胞来模拟血管功能。多孔膜广泛用于创建细胞培养环境。然而，在微通道中对细胞代谢物进行原位实时测量具有挑战性。在这项研究中，开发了一种具有多孔膜电极的新型微流控装置，用于实时原位监测内皮细胞释放的一氧化氮 (NO)。在该系统中，将多孔 Au 膜电极直接放置在细胞下方，用于原位和实时测量作为内皮细胞生物标志物的 NO。首先，对该装置进行了电化学表征，以构建 NO 的校准曲线。接下来，成功定量了人脐静脉内皮细胞在 l-精氨酸刺激下释放的 NO。此外，通过利用微创测量，成功记录了相同样品的培养时间 (以天计) 内 NO 释放的变化。这是首次报道将微流控装置与多孔膜电极相结合，用于内皮细胞的电化学分析。该装置将有助于开发用于实时原位细胞分析的器官芯片技术。
