Open multi-organ communication device for easy interrogation of tissue slices.
Open multi-organ communication device for easy interrogation of tissue slices.
复制标题
开放式多器官通信装置，可轻松询问组织切片。
DOI：
10.1039/d3lc00115f
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发表时间：
2023
期刊：
影响因子：
6.1
通讯作者：
Ross,AshleyE
中科院分区：
文献类型：
--
作者：
Delong,LaurenM;Ross,AshleyE
在这里，我们已经开发了一种开放的多器官通信设备，促进离体器官切片之间的细胞和分子通信。测量器官之间的通信对于理解健康调节机制至关重要，但目前的技术仍然很困难。沿着肠-脑-免疫轴沿着器官之间的通信是肠道内稳态的关键调节器。作为该装置的一种新应用，我们使用了派伊尔集合淋巴结（PP）和肠系膜淋巴结（MLN）的组织切片，因为它们在肠道免疫中很重要;然而，这里可以使用任何器官切片。该设备是使用用于聚二甲基硅氧烷（PDMS）软光刻、PDMS膜和径迹蚀刻多孔膜的3D打印模具的组合设计和制造的。为了验证芯片上器官之间的细胞和蛋白质转移，我们使用荧光显微镜来定量荧光蛋白和细胞从PP到MLN的移动，复制肠道中对免疫刺激的初始反应。对从未处理PP与发炎PP到健康MLN的灌注期间的IFN-γ分泌进行定量，以证明可溶性信号传导分子在芯片上移动。最后，瞬态儿茶酚胺释放测量灌注过程中，从PP MLN使用快速扫描循环伏安法在碳纤维微电极，以证明一种新的应用程序的设备，在通信过程中的实时传感。总的来说，我们展示了一种能够促进可溶性因子和细胞转移的开放式多器官装置，其额外的好处是可用于外部分析技术，如电化学传感，这将提高在体外跨多个器官实时探测通信的能力。
Here, we have developed an open multi-organ communication device that facilitates cellular and molecular communication between ex vivo organ slices. Measuring communication between organs is vital for understanding the mechanisms of health regulation yet remains difficult with current technology. Communication between organs along the gut–brain-immune axis is a key regulator of gut homeostasis. As a novel application of the device, we have used tissue slices from the Peyer's patch (PP) and mesenteric lymph node (MLN) due to their importance in gut immunity; however, any organ slices could be used here. The device was designed and fabricated using a combination of 3D printed molds for polydimethylsiloxane (PDMS) soft lithography, PDMS membranes, and track-etch porous membranes. To validate cellular and protein transfer between organs on-chip, we used fluorescence microscopy to quantitate movement of fluorescent proteins and cells from the PP to the MLN, replicating the initial response to immune stimuli in the gut. IFN-γ secretion during perfusion from a naïve vs. inflamed PP to a healthy MLN was quantitated to demonstrate soluble signaling molecules are moving on-chip. Finally, transient catecholamine release was measured during perfusion from PP to MLN using fast-scan cyclic voltammetry at carbon-fiber microelectrodes to demonstrate a novel application of the device for real-time sensing during communication. Overall, we show an open-well multi-organ device capable of facilitating transfer of soluble factors and cells with the added benefit of being available for external analysis techniques like electrochemical sensing which will advance abilities to probe communication in real-time across multiple organs ex vivo.