FACS analysis and RT-PCR of germ cell specific markers. (A-C) FACS analysis (A) representative histogram, (B) dot plot of one of the six independent experiments (C) bar graph describes mean ± SD of six independent experiments. (D) Immunofluorescence staining of GDF9 in a group of hAECs cultured in medium containing SSS at passage 5 (magnification x20); white arrows, hAECs stained for GDF9; Green color, GDF9; Blue color, DAPI nuclear staining. (E) RT-PCR of C-KIT, DAZL, VASA and ZP3 gene expression in freshly isolated hAECs and in hAECs cultured in medium containing SSS and respectively in standard medium at passage 1 and 5, GAPDH gene expression was used as an internal control. Shown are photographs of the representative bands of the gene products in a 2.5% agarose gel.