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. 2020 Oct 5;10:16459. doi: 10.1038/s41598-020-73177-y

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© The Author(s) 2020

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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Human IPSC-derived Cortical Spheroids Produce Hyaluronan ECM. (A) 10 μm thick cryosections of 90-day-old control spheroids were stained for ECM components. Left to right: merged channels, nuclei marker DAPI (blue), HA as detected by HABP (green), HA synthase HAS2 (red), and HA receptor CD44 (white). Bottom panel highlights a section of the cortical plate at increased magnification. Scale bar of top panel: 100 μm, bottom panel: 20 μm. (B) Graphic illustration of how HA is produced by HAS and interacts with CD44 at the cell membrane. (C) 10 μm thick cryosections of 90-day-old control spheroids were stained for DAPI (blue), HABP (green), neuronal marker DCX (red) and HAS2 (white). Arrows in top left panel highlight the cortical plate. Bottom panel highlights a section of the cortical plate at increased magnification. Scale bar of top panel: 100 μm, bottom panel: 20 μm. (D) 10 μm thick cryosections of 90-day-old control spheroids were stained for DAPI (blue), HABP (green), astrocyte marker GFAP (red) and HAS2 (white). Bottom panel highlights a section of the cortical plate at increased magnification. Scale bar of top panel: 100 μm, bottom panel: 20 μm. Further analysis of cell-type specific expression of HA-ECM components can be found in Fig. S2, while validation of HAS2 immunostaining can be found in Fig. S3.