Fig. S6

Propagation and differentiation culture of SSCs of normal testis. (A) Effects of basal medium and heparin at 50 U/ml on proliferation of dissociated testicular cells of the sox17::egfp transgenics. The gas condition was adjusted to the basal culture medium; in air for L-15 and in 5% CO2 and 10% O2 for DMEM. Bars indicate the average (n=4). (B) Dissociated sox17::egfp testicular cells were cultured for 1, 20, and 31 days under the SSC propagation condition. Upper panels: phase contrast; bottom panels: fluorescent. Arrows indicate GFP-negative differentiated spermatogonia. (C) Sycp3 expression in cells under differentiation culture. SSCs of normal testis that underwent propagation culture for 31 days were transferred onto ZtA6-12 feeders and cultured for 9 days. Sycp3 signals are observed (green). (D) Sycp3 immunostaining of SSC clumps cultured under the SSC propagation condition for 40 days. Cells expressing Sycp3 were observed at the periphery of SSC clumps (green). Scale bars, 50 µm.