Simple SummaryThis study aimed to evaluate whether serum gonadotropin and sex steroid hormone measurement is useful in the differential diagnosis of granulosa cell tumors (GCTs). Serum hormone levels were measured preoperatively in patients who underwent surgery for ovarian tumors (n = 471) and compared in two groups, a GCT group (n = 13) and a group with other histological types of ovarian tumor (non-GCT) (n = 458). Univariate analysis showed that the GCT group had significantly lower serum gonadotropin levels and significantly higher serum sex steroid hormone levels than the non-GCT group. Multivariate analysis revealed that the serum follicle-stimulating hormone (FSH) level was significantly associated with GCT (p = 0.004). Receiver-operating characteristic curve analysis for the diagnosis of GCTs showed that the area under the curve of the serum FSH level was 0.99, with a sensitivity of 100% and a specificity of 98%. Preoperative serum FSH levels are an extremely useful marker for differentiating GCTs from all ovarian tumors.Background: We evaluated whether the serum hormone levels are useful in the differential diagnosis of granulosa cell tumors (GCTs), regardless of menopausal status. Methods: Serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone, estradiol, and progesterone were measured preoperatively in all patients (n = 471) who underwent surgery for ovarian tumors at Chiba University Hospital between 2009 and 2021. These were compared in two groups, a GCT group (n = 13) and a group with other histological types (non-GCT) (n = 458). Results: The GCT group had significantly lower serum LH and FSH (p = 0.03 and p < 0.001, respectively) and significantly higher testosterone, estradiol, and progesterone (p < 0.001, p < 0.001, and p = 0.045, respectively) than the non-GCT group. Multivariate analysis revealed that serum FSH and estradiol were significantly associated with GCT (FSH, odds ratio (OR) = 0.0046, 95% confidence interval (CI) = 0.0026–0.22, p = 0.004; estradiol, OR = 0.98, 95% CI = 0.96–0.998, p = 0.046). Receiver-operating characteristic curve analysis for GCTs showed that the area under the curve of serum FSH was 0.99, with a sensitivity of 100% and a specificity of 98%, when the cutoff level was set at 2.0 IU/L. Conclusions: Preoperative serum FSH level is an extremely useful marker for differentiating GCTs from all ovarian tumors.