This study was conducted to examine the impact of different levels of Insulin hormone in some some characteristics of local cocks’ semen after being cooling for four different periods. this study was executed at the Poultry Research Station/ Agricultural Research Department/ Ministry of Agriculture, for the period of 12^th/Feb to 21^th/ Dec/2022. cocks’ semen was pooled, diluted with extender, and divided equally in to five groups: control treatment (C₁) 0 insulin, (C₂) contained 4 IU/insulin, (C₃) contained 5 IU/insulin., (C₄) contained 6 IU/insulin., and (C₅) contained 7 IU/insulin. The effect of these addition was studied for different cooling periods at 0 ,24 ,48 and 72 hr on ATPase, mitochondrial activity and DNA damage. The results revealed that the addition of insulin hormone were not significant differences in mitochondrial activity, A significant differences were observed on ATPase and DNAdamage when insulin hormone added with high levels within all of the cryopreservation periods.