The objective of this study was to evaluate the characteristics of Korean Native Cattle sperm frozen-thawed with L-cysteine and/or catalase. The semen from bulls was collected by the artificial vagina method, and Triladyl containing 20% egg-yolk and/or L-cysteine (L), catalase (C) and L-cysteine + catalase was added to the diluted semen for cryopreservation. The results showed that sperm viability was significantly higher in the L-cysteine + catalase ($69.49{pm}3.16%$) group than in the control ($60.5{pm}3.94%$) group (p<0.05). Acrosome damage was significantly lower in the L-cysteine ($17.12{pm}1.08%$) group than in the control ($21.46{pm}1.14%$), catalase ($20.54{pm}0.76%$), and L-cysteine + catalase ($19.29{pm}0.65%$) groups (p<0.05). In addition, the level of intact mitochondria in the spermatozoa was significantly higher in the L-cysteine ($58.65{pm}1.39%$) group than in the control ($50.63{pm}2.37%$) group (p<0.05). The hydrogen peroxide level in the frozen-thawed sperm was significantly lower in the L-cysteine ($3.74{pm}1.66%$), catalase ($4.65{pm}1.87%$), and L-cysteine + catalase ($8.11{pm}2.15%$) groups than in the control ($13.22{pm}1.6%$) group (p<0.05). The glutathione level was significantly higher in the L-cysteine ($1.33{pm}0.03%$) group than in the control ($1.08{pm}0.06%$), catalase ($1.05{pm}0.02%$) and L-cysteine + catalase ($1.11{pm}0.03%$) groups (p<0.05). In conclusion, L-cysteine and catalase could protect the membrane of Korean Native Cattle sperm from damage during sperm cryopreservation. Especially, L-cysteine was more effective for keeping acrosomes and mitochondria intactness during sperm cryopreservation.