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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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Retinal differentiation and selection of THY1-positive RGCs using a AAVS1:CAG-P_EGFP hiPSC line. (A) Phase-contrast and brightfield micrograph showing emergence of D28 retinal organoids from differentiating AAVS1:CAG-P_EGFP hiPSCs. Scale bar: 200 μm. (B) Endogenous GFP expression in D56 retinal organoids. Scale bar: 200 μm. (C,D) Immunostaining showing the expression of GFP in all retinal organoid cells. (C) RGCs are identified at D56 according to the co-expression of PAX6 and BRN3A. (D) Co-expression of CRX and recoverin (RCVRN) corresponds to photoreceptors in D130 organoids. Scale bars: 30 μm. (E) RGC characterization in unsorted, THY1– and THY1+ fractions according to the expression of RBPMS, ISLET1, BRN3A, and THY1. Immunostaining for CRX and RCVRN enables photoreceptor identification. Scale bars: 50 μm.