A high-throughput microtissue platform to probe endothelial function in vitro.
A high-throughput microtissue platform to probe endothelial function in vitro.
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DOI：
10.1039/c8ib00111a
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发表时间：
2018-09-17
期刊：
影响因子：
--
通讯作者：
Wood DK
中科院分区：
文献类型：
--
作者：
Crampton AL;Cummins KA;Wood DK
血管内皮的关键作用是作为半渗透屏障，动态调节血液和周围组织之间的溶质通量。现有的在体外定量内皮功能的平台要么是显著的通量受限的，要么忽略了生理相关的细胞外基质（ECM）相互作用，因此不能概括体内功能。利用液滴微流体技术，我们开发了一个可扩展的平台来测量纳升体积的基于ECM的微组织中的内皮功能。在这项研究中，我们描述了我们的高通量的方法来制造内皮涂层胶原微组织，将生理相关的细胞-ECM相互作用。我们发现，内皮细胞具有特征性的形态，表达紧密连接蛋白，并通过基底膜的压实和沉积重塑ECM。我们还使用两种光学模式测量了大分子渗透性，发现细胞层：（1）具有与体内测量相当的渗透性值，（2）对内皮渗透性的生理相关调节剂（TNF-α和TGF-β）有反应。据作者所知，这是第一次对天然ECM的内皮渗透性进行高通量评估（n>150）。此外，该技术与标准细胞培养设备（例如多孔板）兼容，并且可以进一步扩大规模以与自动化液体处理系统和自动化成像平台集成。总的来说，该平台概括了传统transwell插入物的功能，但将应用扩展到高通量研究，并为询问细胞-细胞和细胞-基质相互作用引入了新的可能性。
A critical role of vascular endothelium is as a semi-permeable barrier, dynamically regulating the flux of solutes between blood and the surrounding tissue. Existing platforms that quantify endothelial function in vitro are either significantly throughput limited or overlook physiologically relevant extracellular matrix (ECM) interactions and thus do not recapitulate in vivo function. Leveraging droplet microfluidics, we developed a scalable platform to measure endothelial function in nanoliter-volume, ECM-based microtissues. In this study, we describe our high-throughput method for fabricating endothelial-coated collagen microtissues that incorporate physiologically relevant cell-ECM interactions. We showed that the endothelial cells had characteristic morphology, expressed tight junction proteins, and remodeled the ECM via compaction and deposition of basement membrane. We also measured macromolecular permeability using two optical modalities, and found the cell layers: (1) had permeability values comparable to in vivo measurements and (2) were responsive to physiologically-relevant modulators of endothelial permeability (TNF-α and TGF-β). This is the first demonstration, to the authors’ knowledge, of high-throughput assessment (n>150) of endothelial permeability on natural ECM. Additionally, this technology is compatible with standard cell culture equipment (e.g. multi-well plates) and could be scaled up further to be integrated with automated liquid handling systems and automated imaging platforms. Overall, this platform recapitulates the functions of traditional transwell inserts, but extends application to high-throughput studies and introduces new possibilities for interrogating cell-cell and cell-matrix interactions.
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影响因子：
4.6
作者：
Egawa, Gyohei;Nakamizo, Satoshi;Natsuaki, Yohei;Doi, Hiromi;Miyachi, Yoshiki;Kabashima, Kenji
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