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. 2020 Oct 21;10:17937. doi: 10.1038/s41598-020-73908-1

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Generation of definitive endoderm using defined medium (Step I). (a) Morphological representation of definitive endoderm (DE) differentiated from four different induced pluripotent stem cell (iPSC) lines using conventional or chemically defined, animal origin-free (CD-AOF) medium. Scale bar, 100 µm. (b) Flow cytometric analysis of the CXCR4 expression in DEs differentiated using the conventional or the CD-AOF medium. (c) Gene expression analysis of the DEs differentiated using the conventional or the CD-AOF medium. Error bars represent standard deviation. (d) Immunostaining of the DE for FOXA2 and SOX17. Scale bar, 100 µm.