Figure 5: Appearance of ovarian FLCs induced in differentiated hESCs.

(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.