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. 2019 Dec 30;11(5):983–1000. doi: 10.1007/s12975-019-00773-0

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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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Fig. 1 — Generation of cerebral organoids (COs). a Schematic diagram of COs development. The initial formed embryonic bodies (EBs) in the low-attachment plate at 4 days after induction (DAI) from human embryonic stem cells H1. EBs at 8 DAI with evidence of ectodermal differentiation consisting of brightened surface and relative dark center in the tissue. The healthy EBs showed a smooth surface. After Matrigel embedding for stationary culture of expanding neuroepithelial buds, well-defined polarized neuroepithelium-like structures resembled neural tubes at 15 DAI. Then, cerebral tissues were transferred into the spinning bioreactor for further culture. Here are examples of healthy and failed COs at 30 DAI, respectively. b Immunostaining of SOX2 (green, neural progenitor cells marker) and Tuj1 (red, neurons marker) for cultured cerebral tissues at 15, 30, and 60 DAI. c Immunostaining of COs at 75 DAI with forebrain marker Foxg1 (red) and choroid plexus marker TTR (red). d Immunostaining of COs at 55 days with neural stem cells (Nestin), neurons (Tuj-1), and astrocytes (GFAP). DAPI labels nuclei (blue). All scale bars are as shown