Sexual hormones are essential for the process of spermatogenesis in the testis. However, the effect of maternal genistein (GEN) on the pups' testicular development remain-unclear. Our present study evaluated the effects of supplementing GEN for parental and offspring mice on the reproductive function and growth performance of the male pups. Mothers during gestation and lactation period were assigned to a control diet (CON group), low dose GEN (LGE group) diet (control diet +40 mg/kg GEN), and high dose of GEN (HGE group) diet (control diet +800 mg/kg GEN). Their male offspring underwent the same treatment of GEN after weaning. LGE treatment (40 mg/kg GEN) significantly increased body weights (p < 0.001), testes weights (p < 0.05), diameters of seminiferous tubule (p < 0.001) and heights of seminiferous epithelium (p < 0.05) of offspring mice. LGE treatment also increased serum testosterone (T) levels and spermatogenesis scoring (p < 0.05). However, HGE treatment (800mg/kg GEN) significantly decreased body weights (p < 0.001), testes weights (p < 0.05) and testis sizes (p < 0.001). Furthermore, mRNA expressions of ESR2 (p < 0.05), CYP19A1 (p < 0.001), SOX9 (p < 0.001) and BRD7 (p < 0.001) in testis of mice were increased in the LGE group. Similarly, HGE treatment increased mRNA expressions of ESR2 (p < 0.05) and CYP19A1 (p < 0.001). However, mRNA expressions of SOX9 and BRD7 were decreased significantly in the HGE group (p < 0.001). Meanwhile, higher ratio apoptotic germ cells and abnormal sperms were detected in the HGE group (p < 0.001). In conclusion, exposure to a low dose of GEN during fetal and neonatal life could improve testicular development of offspring mice, whereas, unfavorable adverse effects were induced by a high dose of GEN.