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. 2020 Jun 26;14:538. doi: 10.3389/fnins.2020.00538

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Copyright © 2020 Eura, Matsui, Luginbühl, Matsubayashi, Nanaura, Shiota, Kinugawa, Iguchi, Kiriyama, Zheng, Kouno, Lan, Kongpracha, Wiriyasermkul, Sakaguchi, Nagata, Komeda, Morikawa, Kitayoshi, Jong, Kobashigawa, Nakanishi, Hasegawa, Saito, Shiromizu, Nishimura, Kasai, Takeda, Kobayashi, Inagaki, Tanaka, Makinodan, Kishimoto, Kuniyasu, Nagamori, Muotri, Shin, Sugie and Mori.

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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Schematic procedure of inducing hBSOs and immunohistochemical analysis of 1-month-old hBSOs from hESCs. (A) Schematic procedure of generating human brainstem organoids. SB, SB431542; D, dorsomorphin; I, insulin; P, progesterone; T, transferrin; bF, basic fibroblast growth factor; E, epidermal growth factor; AA, ascorbic acid; cA, cyclic adenosine monophosphate; N, neurotrophin 3; G, glial cell line–derived neurotrophic factor; B, brain-derived neurotrophic factor. Photographs of organoids were taken on days 0, 9, 22, and 28. Bars = 500 μm. (B) Immunohistochemistry of hBSOs at 1-month old from hESCs for the markers of melanocyte (Fontana–Masson, HMB45) and neural crest cell (SOX9). Bars = 100 μm.