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一种用于可视化在流动条件下免疫细胞穿过血脑屏障的硅纳米膜平台。

A silicon nanomembrane platform for the visualization of immune cell trafficking across the human blood-brain barrier under flow.

机构信息

1 Theodor Kocher Institute, University of Bern, Bern, Switzerland.

2 Department of Biomedical Engineering, University of Rochester, Rochester, NY, USA.

出版信息

J Cereb Blood Flow Metab. 2019 Mar;39(3):395-410. doi: 10.1177/0271678X18820584. Epub 2018 Dec 19.

Abstract

Here we report on the development of a breakthrough microfluidic human in vitro cerebrovascular barrier (CVB) model featuring stem cell-derived brain-like endothelial cells (BLECs) and nanoporous silicon nitride (NPN) membranes (µSiM-CVB). The nanoscale thinness of NPN membranes combined with their high permeability and optical transparency makes them an ideal scaffold for the assembly of an in vitro microfluidic model of the blood-brain barrier (BBB) featuring cellular elements of the neurovascular unit (NVU). Dual-chamber devices divided by NPN membranes yield tight barrier properties in BLECs and allow an abluminal pericyte-co-culture to be replaced with pericyte-conditioned media. With the benefit of physiological flow and superior imaging quality, the µSiM-CVB platform captures each phase of the multi-step T-cell migration across the BBB in live cell imaging. The small volume of <100 µL of the µSiM-CVB will enable in vitro investigations of rare patient-derived immune cells with the human BBB. The µSiM-CVB is a breakthrough in vitro human BBB model to enable live and high-quality imaging of human immune cell interactions with the BBB under physiological flow. We expect it to become a valuable new tool for the study of cerebrovascular pathologies ranging from neuroinflammation to metastatic cancer.

摘要

在这里,我们报告了一种突破性的微流控人体外脑血管屏障 (CVB) 模型的开发,该模型具有干细胞衍生的类脑内皮细胞 (BLEC) 和纳米多孔氮化硅 (NPN) 膜 (µSiM-CVB)。NPN 膜的纳米级薄度与其高通透性和光学透明度相结合,使其成为用于组装具有神经血管单元 (NVU) 细胞成分的体外血脑屏障 (BBB) 微流控模型的理想支架。由 NPN 膜分隔的双室装置在 BLEC 中产生紧密的屏障特性,并允许用周细胞条件培养基替换周细胞共培养物。µSiM-CVB 平台具有生理流动和卓越的成像质量的优势,可在活细胞成像中捕获 T 细胞穿过 BBB 的多步迁移的每个阶段。µSiM-CVB 的小体积<100µL 将能够在体外研究具有人类 BBB 的罕见患者来源的免疫细胞。µSiM-CVB 是一种突破性的体外人类 BBB 模型,可在生理流动下实现人类免疫细胞与 BBB 相互作用的实时和高质量成像。我们预计它将成为研究从神经炎症到转移性癌症等脑血管疾病的有价值的新工具。

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