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. 2021 Jul 27;12:4535. doi: 10.1038/s41467-021-24781-7

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© The Author(s) 2021

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 Drawing of the steps in the Sasai and 3D-RET retinal organoid protocol. The number of retinal organoids produced from a 96-well dish is indicated for representative lines for each protocol. Red arrows indicate media changes. B, C Micrograph of representative retinal organoids using the Sasai and 3D-RET protocols in a side-by-side comparison for SJRB-iPSC-4. All organoids from a 96-well dish were analyzed. Arrows indicate retina organoids and (*) indicates cystic structures that are common in the Sasai protocol. D Micrographs of dissociated cell immunofluorescence of retinal organoids and a dot plot showing the percentage of recoverin immunopositive cells from individual retinal organoids (n = 8). MG Matrigel, SAG smoothened agonist, RMM retinal maturation medium, FBS fetal bovine serum, CHIR GSK3 inhibitor, ec23 retinoic acid analog. Scale bars: B, C, 100 μm. D, 5 μm.