Department of Chemical Engineering, Hongik University, Seoul, 04066, Republic of Korea.
School of Electrical and Electronic Engineering, Yonsei University, Seoul, Korea 03722.
Lab Chip. 2024 Sep 24;24(19):4581-4593. doi: 10.1039/d4lc00490f.
The gut communicates with the brain in a variety of ways known as the gut-brain axis (GBA), which is known to affect neurophysiological functions as well as neuronal disorders. Exosomes capable of passing through the blood-brain-barrier (BBB) have received attention as a mediator of gut-brain signaling and drug delivery vehicles. In conventional well plate-based experiments, it is difficult to observe the exosome movement in real time. Here, we developed a microfluidic-based GBA chip for co-culturing gut epithelial cells and neuronal cells and simultaneously observing exosome transport. The GBA-chip is aimed to mimic the situation of convective flow in blood vessels and convective and diffusive transport in the tissue interstitium. Here, fluorescence-labeled exosome was produced by transfection of HEK-293T cells with CD63-GFP plasmid. We observed in real time the secretion of CD63-GFP-exosomes by the transfected HEK-293T cells in the chip, and transport of the exosomes to neuronal cells and analyzed the dynamics of GFP-exosome movement. Our model is expected to enhance understanding of the roles of exosome in GBA.
肠道通过被称为肠道-大脑轴（GBA）的多种方式与大脑进行通讯，已知它会影响神经生理功能和神经元紊乱。能够穿过血脑屏障（BBB）的外泌体作为肠道-大脑信号转导的介质和药物传递载体受到关注。在传统的微孔板实验中，很难实时观察外泌体的运动。在这里，我们开发了一种基于微流控的 GBA 芯片，用于共培养肠上皮细胞和神经元细胞，并同时观察外泌体的运输。GBA 芯片旨在模拟血管中对流的情况以及组织间隙中的对流和扩散运输。在这里，通过用 CD63-GFP 质粒转染 HEK-293T 细胞产生荧光标记的外泌体。我们实时观察了转染的 HEK-293T 细胞在芯片中分泌 CD63-GFP-外泌体，并观察到外泌体向神经元细胞的运输，分析了 GFP-外泌体运动的动力学。我们的模型有望增强对外泌体在 GBA 中作用的理解。
