Fibroblast growth factor 8 (FGF8), acting through the fibroblast growth factor receptor 1 (FGFR1), has an important role in the development of GnRH neurons. We hypothesized that FGF8 regulates human GnRH neuron differentiation in a time- and dose-dependent manner via FGFR1. Human pluripotent stem cells were differentiated to GnRH neurons with ten days of dual-SMAD inhibition, followed by FGF8 (25 ng/ml, 50 ng/ml or 100 ng/ml for ten days; or 100 ng/ml for two, four, six, and ten days), and Notch inhibition for five days. FGF8 induced expression of GNRH1 in a dose-dependent fashion and the duration of FGF8 exposure correlated positively with GNRH1 expression (P<0.05, Rs=0.49). However, two days of FGF8 (100 ng/ml) induced expression of its target genes such as FOXG1, ETV5, and SPRY2, and continued FGF8 induced dynamic expression of several genes. During FGF8 exposure, FGFR1 localized to the cell surface, and its specific inhibition with PD166866 reduced GNRH1 expression (P<0.05). In neurons, FGFR1 also localized to the nucleus. Our results suggest that dose- and time-dependent FGF8 signaling via FGFR1 is indispensable for human GnRH neuron ontogeny.