Eurasian perch is an important fish species for European aquaculture diversification, but the quality of reproduction still remains one of the main limitations for further industry development. In particular, the optimal condition to obtain the best quality of sperm is poorly understood. The aim of our study was to measure the possible effects of two experimental rearing temperatures (6?°C and the conventionally used 12?°C) and of hormonal stimulation, on the motility parameters (pMOT, VCL, VSL, LIN, ALH, BCF), osmolality and fertilizing capacity of Eurasian perch sperm at the end of the reproductive cycle. A prior untested, large-scale (5?mL cryotube and Polystyrene box) cryopreservation method was implemented using fresh sperm obtained from the two above mentioned temperature groups. Males were injected with 100?μg body weight kg−1 sGnRHa. No significant difference was recorded between the two rearing temperatures and between the saline control and sGnRHa treated groups on the different features of sperm quality. A similar fertilization rate was monitored in all sGnRHa treated (6?°C: 69?±?13%, 12?°C: 81?±?11%) and saline control groups (6?°C: 79?±?10%, 12?°C: 87?±?4%). Correspondingly, no significant difference in hatching rate was observed in the sGnRHa injected (6?°C: 27?±?9%, 12?°C: 40?±?20%) and saline control (6?°C: 35?±?18%, 12?°C: 36?±?7%) males. However, a notable negative effect of freezing process on sperm movement was observed following thawing in both temperature groups. No significant difference in the motility parameters was measured between the two temperature groups following large-scale cryopreservation. Furthermore, a similar result was observed in the fertilizing capacity (6?°C: 79?±?10%, 12?°C: 75?±?8) of thawed sperm as well as in the hatching rate (6?°C: 52?±?13%, 12?°C: 46?±?19%). Our results indicate that fresh Eurasian perch sperm can tolerate a reduced rearing temperature following hormonal treatment. The adopted large-scale cryopreservation method could be used efficiently in the future for the fertilization of large amounts of Eurasian perch eggs following a precise standardization process.