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. 2018 Feb 12;8:2814. doi: 10.1038/s41598-018-21098-2

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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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3D Matrigel promotes aggregation and upregulates expression of markers of TM cells, ESCs and NC. P3 cells cultured on 2D Matrigel in MESCM + 5% FBS were reseeded in 3D Matrigel with or without Noggin before morphological analysis by phase contrast microscopy (C, scale bar: 20 µm), immunostaining to Oct4, Sox2, Nanog, Myc, or KLF4 (C, nuclear counterstained by Hoechst 33342, scale bar: 20 µm), and qRT-PCR for TM markers (A) and ESC and neural crest markers (B, n = 3, **P < 0.01 and ***P < 0.001) by setting the expression level for 2D Matrigel as the control.