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. 2018 Aug 27;8:12865. doi: 10.1038/s41598-018-31256-1

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© The Author(s) 2018

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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Modulation of rostro-caudal and dorso-ventral axis specification induces the development of PHOX2B-expressing NCCs from hPSCs. (a) A diagram of the culture conditions for the modification of rostro-caudal and dorso-ventral axis specification of hPSCs. (b) A heat map image showing the percentage of eGFP⁺ cells on day 10 of differentiation using KhES1 PHOX2B::eGFP under various conditions. (c) Representative FCM plots of day 10 KhES1 PHOX2B::eGFP-derived aggregates under the indicated conditions (i–iv) of (b). (d) RT-PCR analyses for PHOX2B, SOX1, PAX6, SOX10, FOXD3, HOXB1, HOXB2, HOXB4, HOXB6, HOXB8 and HOXC9 in day 10 aggregates under conditions (i–iv). The right diagram shows the expression pattern of HOX genes in the rhombomere (r1–8) and the spinal cord (cervical and thoracic) region. SB = SB431542, CHIR = CHIR 99021, RA = retinoic acid, Pur = Purmorphamine, BMP = BMP4, NT = neural tube, NCC = neural crest cell, HB = hindbrain, SC = spinal cord.