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. 2017 Aug 15;7:8299. doi: 10.1038/s41598-017-08266-6

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© The Author(s) 2017

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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Cell lineage analysis of blastocysts. Immunoconfocal fluorescence images of representative MZ twin blastocysts, co-stained for cell lineage-specific markers: CDX2 for TE (A1,B1), SOX17 for pEnd (A2,B2) and NANOG for EPI (A3,B3). Microscope objective 20X, 0.75 N.A. Pairs were scored as either “concordant” (A) or “discordant” (B) based on the “9-2-2 criterion” (see main text). The proportions of concordant and discordant pairs were similar irrespective of culture medium, ovarian stimulation, co-culture, sperm-entry point and embryo conformation at the four-cell stage (tetrahedral, T; planar, P; open chain, O). The error bars associated with the proportions are standard deviations. (C). Linear correlation analysis (Pearson’s r) of the cells counted in the lineage compartments of each MZ pair shows that the main contributor to the discordance was the EPI lineage (D).