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. 2023 Jun 14;21:132. doi: 10.1186/s12964-023-01159-4

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© The Author(s) 2023

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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — ICC staining of neurovascular networks on chip at day 14 timepoint. a Tilescan images showing the BMSC and ASC multicultures. Staining for a vascular differentiation marker (CD31, red) showed consistent and repeatable vasculature formation throughout the microfluidic chip in the BMSC and ASC multicultures. Staining for neuronal markers (MAP-2 + βtubIII, green) showed that a neuronal network had also formed throughout the hydrogel in both multiculture formats. Scale bar is 500 µm. b Close-up image of the BMSC multiculture. c Close-up image of the ASC multiculture. d-e Confocal 3D rendering of the multicultures showing neurons (MAP-2 + βtubIII, green) in the connections to both HUVECs (CD31, red) and mural cells (α-SMA, gray) in both the d BMSC and e ASC multicultures. Pink arrowheads indicate the neuronal connections to ECs and mural cells. Scale bar is 10 µm. The illustrations were created using Biorender.com