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. 2017 Jul 17;7:5575. doi: 10.1038/s41598-017-06008-2

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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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Upon removal of FGF4, mouse TS cell cultures were initially proliferative but increased in number only until day 4 of differentiation (A). Sca-1⁺ trophoblast cells isolated from non-proliferating, differentiated TS cultures and plated at decreasing densities in conditions to support TS proliferation formed colonies however Sca-1⁻ and non-sorted, differentiated TS cells did not (B). When these cells were compared to the Sca-1⁻ subpopulation of trophoblast cells by qRT-PCR, Sca-1⁺ positive cells displayed higher expression of Sca-1 and the trophoblast stem and progenitor markers, Esrrb and Epcam, Rhox4, respectively, while Ascl2 was not different (C).