As a result of experimental researches it has been established that the optimization of parameters at the development of new mediums helps to reduce the cryogenic risks in the preservation of the genetic resources of the rooster. The best results in the restoring of gametes physiological parameters of after cryopreservation in maltose-petumozid-arginineglycerin medium and defrozen in isotonic sodium citrate solution are achieved at the dilution of semen 1:3, refrigeration at 2-4°C during 15 minutes and freezing for a duration of 2 minutes. The use of such cryobiological parameters allowed us to obtain similar results when using maltose-arginine-glycerol medium as in the control group. At the same time, the additional use of petumozide in the composition of experimental medium increases the mobility of thawed gametes of cock by 11.4%. The effectiveness of the developed medium was tested and in the production conditions. As a result, it was established that the hatchability of chicks reaches 68.8 ± 6.69 against 41.7 ± 7.12%, which is 27.1% higher in favor of the experimental variant. The results of laboratory and industrial experiments allow to recommend developed medium for cryopreservation of semen of the rooster and use in breeding practice.