The objectives of this study were to determine the effect of different cryoprotectants and sperm densities for longterm storage of orange mud crab, Scylla olivacea spermatozoa. Spermatozoa were obtained by homogenizing the spermatophores using a glass homogenizer in an ice-bath followed by centrifugation at 4°C. Spermatozoa were then suspended in calcium-free saline (Ca-F saline) containing 5% of the following cryoprotectants: Glycerol, dimethyl sulfoxide (DMSO) and methanol. Sperm which vibrated and rotated were counted as live during sperm viability assessment. Samples of spermatozoa were cooled to -196°C by two-step freezing, first to -80°C and then by plunging into liquid nitrogen (LN). Spermatozoa were gradually cooled at 1°C/min. Thawing was carried out in a 30°C water bath for 2 min. This yielded live sperm after storage in LN for 30 days. The best sperm viability was obtained from a density of 108 cells per mL in DMSO. There was no significant difference (p>0.05) among cryoprotectants toward sperm viability. However, sperm viability was significantly affected (p>0.05) by cell densities. In conclusion, DMSO gave the best protection to sperm cells of S. olivacea, but the effectiveness of DMSO as a cryoprotectant is influenced by sperm density.