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. 2015 Mar 6;24(13):1536–1545. doi: 10.1089/scd.2014.0410

FIG. 1.

FIG. 1.

Primordial germ cells (PGCs) enter the gonad hypomethylated and remain hypomethylated over the next 3 days in vivo and ex vivo. (A) Phase-contrast image of an E10.5 aorta-gonad-mesonephros (AGM). Black arrows point to the genital ridge, (m) refers to mesonephros. (B) Histology of an E10.5 AGM. Dotted line separates the genital ridge (black arrow) from the mesonephros (m). (C) Immunofluorescence of E10.5 AGM for stage-specific embryonic antigen-1 (SSEA-1)-positive PGCs (green), nuclei are detected using DAPI (blue), white star refers to PGCs outside of the genital ridge. Dotted line separates the genital ridge from the mesonephros. (D) Phase-contrast image of an AGM cultured for 3 days. Black arrows point to the genital ridge, m refers to the mesonephros. (E) Histology (left) and immunofluorescence (right) of a female (XX) AGM cultured for 5 days (top) and male (XY) genital ridge (bottom). Bottom panel includes high-power images (63×) of the respective male section. Mvh-positive PGCs (red), Amh-positive sertoli cells (green), and DAPI nuclei (blue). (F) Immunofluorescence of E10.5 AGMs for Mvh-positive PGCs (red) and 5-methyl cytosine (5mC) (green). (G) Days 1–3 AGM organ culture for Mvh-positive PGCs (red) and 5mC (green). (H) E11.5–E13.5 genital ridges. All images were obtained using at least n=3 independent biological replicates of gonads or AGMs.