In this study, we used flow a cytometric assay to evaluate plasma membrane integrity and mitochondrial activity in post-thawed sperm that was supplemented with ginsenoside-Rg1. Varying concentrations of ginsenoside-Rg1 (0, 25, 50 and 100 μM/ml) were used in the extender during cryopreservation to protect the DNA of thawed sperm, thereby increasing the viability and motility rate as evaluated using a computer-assisted sperm analysis (CASA) method. The results derived from CASA were used to compare the fresh, control, and ginsenoside-Rg1 groups. Sperm motility and the number of progressively motile sperm were significantly (p<0.05) higher in the 50 μM/ml ginsenoside-Rg1 group (61.0±4.65%) than in the control (46.6±7.02%), 25 μM/ml (46.2±4.76%), and 100 μM/ml ginsenoside-Rg1 (52.0± 1.90%) groups. However, the velocity distribution of post-thawed sperm did not differ significantly. Membrane integrity and MMP staining as revealed using flow cytometry were significantly (p<0.05) higher (91.6±0.82%) in the 50 μM/ml ginsenoside-Rg1 group than in the other groups. Here, we report that ginsenoside-Rg1 affects the motility and viability of boar spermatozoa. Moreover, ginsenoside- Rg1 can be used as a protective additive for the suppression of intracellular mitochondrial oxidative stress caused by cryopreservation.