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. 2016 Jun 30;6:28218. doi: 10.1038/srep28218

Figure 3. Isolation and characterization of human faFGSCs from follicular aspirates.

Figure 3

(A) Representative images of DDX4+ cells from FAs after 2 weeks of culture. The white arrowheads indicate the magnetic beads. (B) The DDX4+ cells were confirmed by immunocytochemical analysis with the antibody against C-terminal of DDX4. (C) A representative morphology of mitotic faFGSCs. (D) A representative morphology of faFGSCs at passage 4 (p4). (E) A representative morphology of faFGSCs at passage 30 (p30). (F) RT-PCR analysis human ovarian tissues (28 years old), faFGSCs (cell line 1# and 2#), and srFGSCs. Lane 1, 100-bp DNA markers. –RT, PCR of RNA sample without reverse transcription. GAPDH is sample loading control. (G) The faFGSC line was detected by immunofluorescence analysis with the antibodies against DDX4, OCT4, IFITM3, BLIMP-1, and DAZL. (H) Karyotype analysis of the faFGSC line by G banding showing normal 46, XX and the distribution of metaphase spreads with different chromosome numbers (right). n = 59. (I) The faFGSC line was detected by immunofluorescence analysis with the antibody against GFRA1. (J) Immunohistochemistry staining of EGFP-positive cells (EGFP+, brown) surrounded by smaller EGFP− cells in adult human ovarian cortical tissues injected by EGFP+ faFGSCs and xenografted into nude female mice for 2 weeks. The nuclei were counterstained with hematoxylin (blue). Full-length gels in F are presented in Supplementary Fig. 5. N.C. in (B,G,J) are omission of the primary antibody. Scale bars: 20 μm (A–C), 50 μm (G, I,J), 100 μm (E), 200 μm (D).