In order to establish Centropristis striata sperm cryopreservation method, this paper used a computer-assisted sperm motion analysis system(CASA)，and selected six cryoprotectants,such as glycerin(GLY), dimethyl sulfoxide(DMSO), propylene glycol(PG), ethylene glycol(EG), dimethyl sulfoxide(DMA), methanol(METH) at four concentrations (5%, 10%, 15% and 20%, V/V) in HBSS to find the influence of C. striata sperm cryopreservation. Results showed that the best cryopreservation protocol for C. striata was 15% PG as antifreeze protective agent in HBSS with step cooling scheme. After 37°C water bath thawed, motility of the sperm reached (93.1±0.9)%, which was higher than other combination groups and showed no significant difference with fresh sperm (P>0.05).The group of 15% PG performed best in average linear velocity, average velocity, and average path velocity at (88.3±0.3) μm/s, (76.2±0.5) μm/s, and (86.7±0.7) μm/s, which showed no significant difference with fresh sperm (P>0.05). In different cryoprotectants with different concentration, the motility of 15% PG within 1 min had no significant difference with fresh semen. Therefore, 15% PG was the best cryoprotectant for C. striata and could be applied in practice.