The ability to transport superior male genetics between rabbitries is essential, thus improvements to the rabbit meat industry are required in Australia. During transportation, sperm viability must be maintained to ensure successful conception. This study assessed different cryoprotectants and their ability to maintain sperm motility and viability post-thaw. In experiment 1, three ejaculates were collected from three individual bucks (n=9) and diluted 1:10 with one of four penetrating cryoprotectant concentrations (1-4; 3.5% DMSO, 1.5% acetamide, 3.5% DMSO + 1.5% acetamide and 1.75% DMSO + 0.75% acetamide, respectively), all containing 17% egg yolk in a standard tris-citric acid-glucose (TCG) diluent. Diluted samples were stored in 0.25 mL straws, slow cooled to 5 °C over 2.5 h and then frozen in liquid nitrogen. Sperm motility was assessed using CASA, and acrosome integrity and plasma membrane integrity were evaluated using flow cytometry. These parameters were assessed immediately after collection, and at 0, 2 and 4 h post-thaw. Results showed Diluent 1 maintained significantly higher total motility (P<0.001; 17.78 ± 4.09 at 4 h) and sperm viability (P<0.001; 53.46 ± 5.31 at 4 h) over each time point post-thaw, when compared to Diluents 2 and 4. In experiment 2, Diluent 1 was further optimised by assessing the cryoprotective effects of egg yolk versus extracted low density lipoproteins (LDL) (A-D; 17% egg yolk, 17% LDL, 9% egg yolk and 9% LDL, respectively). Diluent A had significantly higher total motility (P=0.01; 21.33 ± 2.40 at 4 h) and sperm viability (P<0.001; 62.70 ± 3.56 at 4 h) than other diluents at each time point post-thaw. Overall, the use of 3.5% DMSO in combination with 17% egg yolk in a TCG diluent improved the post-thaw survival of rabbit sperm compared with the other diluents tested.