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. 2020 Oct 27;8:585675. doi: 10.3389/fcell.2020.585675

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Copyright © 2020 Rabesandratana, Chaffiol, Mialot, Slembrouck-Brec, Joffrois, Nanteau, Rodrigues, Gagliardi, Reichman, Sahel, Chédotal, Duebel, Goureau and Orieux.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Selection of hiPSC-derived RGCs from cryopreserved retinal organoids. (A) Schematic diagram illustrating the protocol for the selection of THY1-targeted hiPSC-derived RGCs from cryopreserved hiPSC-derived retinal organoids. (B) Phase-contrast and brightfield micrograph and immunostaining for RGC markers, βIII-tubulin, BRN3A, PAX6, RBPMS, or THY1 in unsorted or THY1-sorted cells from dissociated freeze–thawed organoids (cell nuclei staining with DAPI in blue). Scale bars: 50 μm. (C) Quantitative analysis by flow cytometry of living cells (YOPRO negative) on unsorted, THY1-negative, and THY1-positive fractions, 1 week after plating, of dissociated cells derived from freeze–thawed retinal organoids (min to max; N = 3 differentiation; n = 380 organoids; *p < 0.05, two-way ANOVA followed by Tukey’s multiple comparison test).