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. 2017 Dec 19;11:409. doi: 10.3389/fncel.2017.00409

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Copyright © 2017 Perny, Ting, Kleinlogel, Senn and Roccio.

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) or licensor 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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3D in vitro formation of non-neural ectoderm. (A) Schematic of the differentiation protocol. PP, pluripotent; DE, definitive ectoderm NNE, non-neural ectoderm; PPE, pre-placodal ectoderm; OEPD, otic epibranchial placode domain; HC, hair cells; OSN, otic sensory neurons (OSNs). (B) Morphological changes during the first 3 days of differentiation are visualized by brightfield microscopy. Scale bar 100 μm. (C) Immunostaining of day 5 (d5) organoids for Sox2 and AP2. (D) Immunostaining of day 5 organoids for AP2 and Ecad. 3D maximum projection (3D) and single stack (top and middle) are shown for the acquired confocal stacks. Scale bar 100 μm. (E) Magnified view of the boxed area in D, showing the formed AP2⁺, Ecad⁺ outer epithelium. Scale bar 20 μm.