Abstract: The use of frozen semen in rabbits is greatly limited due to its low fertility rates. The current study aimed to evaluate the effect of gelatin addition to semen extender on motility, livability, abnormality and acrosome integrity of rabbit spermatozoa during freezing process and on fertility after Artificial Insemination (AI). Pooled semen collected from bucks (n = 15) belonging to a line selected by Animal Production Research Institute, Egypt (APRI). Semen was processed in tris-buffer extender with gelatin addition at levels of 0, 1, 2 and 3% (g/100 mL extender) at a rate of 1:5, and frozen in liquid nitrogen. Results showed that when compared with control semen, 2% gelatin addition had positive (p<0.05) effect on percentages of motility, livability, abnormality and intact acrosome o f spermatozoa in post-diluted (73 vs. 65.5%; 73.9 vs. 66.9%; 12.2 vs. 15% and 81 vs. 70%), post-equilibrated (71 vs. 60%; 71.7 vs. 60.4%; 12.6 vs. 15.1% and 80.2 vs. 67.5%) and post-thawed (54.5 vs. 31%; 54.6 vs. 31.4%; 18 vs. 21.4% and 71.4 vs. 56%) semen, respectively. Semen with 2% gelatin yielded the highest (p<0.05) kindling rate (75%) and litter size (6.93) as compared to control semen (65% and 6.08/doe), respectively. Adding 2% gelatin to tris-buffer extender of APRI rabbit semen enhanced freezing ability, efficiency and fertility of spermatozoa in thawed semen.