Semen extenders containing egg yolk as a cryoprotectant may pose hygienic risks and are difficult to standardize. Although a new generation of semen extenders free of animal ingredients is available, egg yolk-containing extenders are still widely used for cryopreserving semen. The aim of the present study was to compare the effect of using soy lecithin-based extenders, Biociphos and Bioxcell, and egg          yolk-based extender on buffalo spermatozoa freezability and fertilizing potentials. Extension of buffalo bull semen in the Bioxcell and the Biociphos extenders significantly increased (P<0.01) the post-thaw sperm motility (61.67 and 60.00%, respectively) and the viability index (130.83 and 120.00, respectively) compared to semen that extended in the TRIS-egg yolk extender (38.33% and 88.33, respectively).  Assessment of the post-thaw acrosomal integrity showed significant differences (P<0.01) between extenders (11.00, 11.33 and 19.00 %, for Bioxcell, Biociphos and TRIS-egg yolk extenders, respectively). In vitro fertilization results revealed that, extension of buffalo bull semen in the Bioxcell and Biociphos extenders significantly increased (P<0.05) the in vitro fertilization rate (65.08 and 63.24%, respectively) and boosted the ability of cleaved oocytes to develop to the morula stage (23.94 and 22.89%, respectively) compared to semen that extended in the TRIS-egg yolk extender (41.07 and 9.62%, respectively). Field trials revealed that, no significant differences (P>0.05) were detected between the extenders for the non-return rates. We suggest that consistent with quality standards that should be required for cryoprotectant media, soy lecithin-based diluents might be the best choice as a buffalo semen extender in the future.