Next, we applied medium B which consisted of Advanced DMEM/F12 (1:1 medium) supplemented with 100 ng/mL EGF, 10 μM Y27632, 5 μM SB202190, 10 mM nicotinamide, 500 nM A-83-01, 2% (v/v) B27, 1% (v/v) Glutamine and 5% (v/v) conditioned medium containing Wnt3A, RSPO1 and Noggin to the next nine ovarian tumor specimens from HOV31T to HOV39T including one germ cell tumor. The gene discussed is RSPO1; the disease is testicular germ cell tumor.