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. 2022 Feb 2;8:47. doi: 10.1038/s41420-022-00845-5

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© The Author(s) 2022

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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Fig. 2 — A A schematic overview of the procedures for generating BOs from hPSCs using the STEMdiff™ Cerebral Organoid Kit. B Representative images of hPSC-derived cells at the corresponding stages. C Representative images of the ventricular zone-like structure formed by new-born neurons (Tuj1⁺) and neural progenitor cells (SOX2 and PAX6⁺) in BOs at day 40. Scale bar: 100μm. D Representative images of the positive staining regions for forebrain (FOXG1) and hindbrain (PAX2) markers respectively. Scale bar: 100 μm. E Representative images showed the immunostaining of the six layers of cortical neurons at day 60. Scale bar: 100 μm. F Confocal images showed the positive staining regions for the astrocyte markers GFAP, and the expression of different subtypes of functional neuron markers, such as TH, 5-HT and vGLUT2. Scale bar: 100 μm.