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. 2017 Dec 1;7:16757. doi: 10.1038/s41598-017-16727-1

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© The Author(s) 2017

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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Effect of locally sustained release of SEMA3A (200 ng/ml) from hydrogels placed in front of attached OVs. Locally sustained release of SEMA3A using hydrogel inhibited anterior axonal growth of hESC- and hiPSC-derived RGCs ((a) and (d), respectively, magnification 40x). In magnified image, most axons are clearly repelled by SEMA3A release ((b) and (e), magnification 100x: squares in (a) and (d), respectively). The filopodia of hESC- and hiPSC-derived RGCs ((c) and (f), respectively) have collapsed owing to SEMA3A release, as evident from phalloidin and GAP43 staining. The assessment from D27 is performed in RMM supplemented with 1.0% FBS and 100 ng/ml BDNF. Each experiment was repeated at least five times. Squares in (a) and (d) correspond to (b) and (c), respectively. Scale bars in (a) and (d), 100 μm. Scale bars in (b) and (e), 40 μm. Scale bars (c) and (f), 10 μm.