Fig. 5

Induction of SSCLCs from pDFP-derived PGCLCs. A Representative pictures of day 7 and 14 aggregates of eGFP-PGCLCs and somatic testicular cells; at 14 days, testicular organoids showing structures resembling testis cords containing eGFP-cells are recognizable. Pictures were taken using an eclipse microscope (10x, Nikon TE2000, Japan). Scale bars, 50 μm. B Expression of the canonical SSC marker PLZF and GFRα1 in eGFP-cells isolated from testicular organoids after 14 days of culture. Pictures were taken using a confocal microscope (20x, Leica, TCS SP5 II, Germany). Scale bars, 20 μm. C Meiotic chromosome spreads of testicular tissues from 8 dpp testes of CAG/eGFP mice and testicular organoids containing eGFP-SSCLCs after 8 days of culture stained with antibodies against meiotic proteins; note that positive staining for SYCP3, γH2AX, and RAD51 was observed in all samples while positive cells for SYCP1 and MLH1 were observed only in the control samples from 8 dpp testes of CAG/eGFP mice. Pictures were taken using a confocal microscope (20x, Leica, TCS SP5 II, Germany). Scale bars, 20 μm