Spermatogenesis is a complex process producing haploid spermatozoa, and the formation of lipid droplets (LDs) within Sertoli cells is critical to maintaining normal spermatogenesis. However, the utilization of LDs within Sertoli cells is still largely unknown. In the present study, proliferation of spermatogonial cells had begun in May, whereas the meiotic cells occurred predominately in July and majority of spermiogenic cells were observed in the seminiferous tubules in October. However, TEM and Oil Red O staining demonstrated that a larger number of LDs had accumulated within the Sertoli cells in May compared to that in October. There were several LDs attached to the isolation membrane/phagophore, suggesting that the LDs may be a source of endogenous energy for the biogenesis of autophagosomes. The LDs were enclosed within the autophagosomes in May, whereas, autophagosomes and mitochondria were directly attached with large LDs within the Sertoli cells in October. Furthermore, immunohistochemistry results demonstrated the stronger localization of LC3 on the Sertoli cells in May than in October. This study is the first to provide clear evidence of the two different modes of lipophagy for lipid consumption within Sertoli cells, which is a key aspect of Sertoli germ cell communication during spermatogenesis.