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. 2021 Jan 26;11:2281. doi: 10.1038/s41598-021-81933-x

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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(A,B) Schematic overview of the microfluidic chip. (A) The open top curved channel (blue) is separated from the straight bottom channel (yellow) by a polyester membrane (5 µm pores). (B) Cross-section connected channels. (C,D,E) neurovascular unit (NVU)-on-a-chip microfluidic device. (C) Open-top microfluidic chip in a 6-well plate, in the bottom channel (yellow) endothelial cells can be cultured, while the open top channel (blue) is used for neuronal differentiation. (D) Staining performed on endothelial cell monoculture on bottom channel of the microfluidic chip on a polyester membrane. Based on the staining pattern, a monolayer was present. Blue: Nuclei; Red: F-actin. (E) Staining performed on neurons differentiated in the top channel of the microfluidic chip on a polyester membrane. Blue: Nuclei; Green: Microtubule Associated Protein 2 (MAP2); Red: Synapsin-1/2 (SYN1/2). (F): Staining performed on neurons co-cultured with astrocytes differentiated on a 24-well plate. Blue: Nuclei; Green: MAP2; Red: Glial fibrillary acidic protein (GFAP), a 1:1 ratio of neurons and astrocytes is visible. (G): Staining performed on neurons differentiated on a 24-well plate. Blue: Nuclei; Green: MAP2; Red: SYN1/2. Scale bars, 50 µm.