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. 2017 Jun 12;8:15680. doi: 10.1038/ncomms15680

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Copyright © 2017, The Author(s)

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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(a) Schematic timelines and simplified protocols for the differentiation from the undifferentiated hESC stage to the appearance of FLCs. (b) Phase-contrast image of a differentiated hESC culture without inducers at day 11. (c) Phase-contrast image of a differentiated hESC culture with the inducers described in a at day 11. (d) Magnified phase-contrast images of four different FLCs appearing at day 11 in the induced culture. (e) Stereo-microscope image of a differentiated hESC culture with inducers at day 11. (f) Stereo-microscope image of a group of FLCs collected from a differentiated hESC culture with inducers at day 11. Immunofluorescent staining of ovarian follicle markers ZP2 (g), NOBOX (h) and AMH (i) on the culture plate with FLCs. (j) VASA-eGFP reporter cells specifically expressed eGFP in cells that became FLCs. P.C.: phase contrast. Bar, 500 μm in b,c,e,f; 100 μm in d; 50 μm in g,h,i,j.