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. 2019 Jul 30;9:11055. doi: 10.1038/s41598-019-47444-6

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© The Author(s) 2019

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Co-Culture of microglia-like cells with ventral, dorsal or ventral/dorsal organoids: BrDU and Ca²⁺ transient assays. Spatial cell distribution of microglia-like cells in co-culture. Microglia-like cells were labeled with microdevices. (A) Overlay of phase contrast images with fluorescent images to show the integration of microglia-like cells within dorsal spheroids, ventral spheroids; and ventral/dorsal spheroids. Red arrows point to the microglia inside the spheroids. Scale bar: 100 μm. (B) BrdU assays for cell proliferation. (i) Fluorescent images of BrdU⁺ cells. Scale bar: 100 μm. (ii) Quantification of BrdU⁺ cells. (C) Ca²⁺ transient assays (induced by ADP). (i) Quantification of Ca²⁺ fluorescent signals; (ii) Fluorescent images of Ca²⁺ transient expression. Scale bar: 100 μm. *Indicates p < 0.05 for the different test conditions.