To our knowledge, the roles of kaempferol in semen extension or cryobiology haven’t been discovered until now. Furthermore, the intracellular signals triggered by kaempferol in buffalo spermatozoa during semen processing haven’t been reported. So, the current study intended to clarify the role of kaempferol during the cryopreservation of buffalo spermatozoa. Six buffalo bulls were adopted for semen collection. Two consecutive ejaculates were collected from each buffalo bull weekly using an artificial vagina. The ejaculates were pooled to eliminate variability. Semen extension was done using the Tris-based diluent supplemented with Kaempferol at concentrations of 5, 10, 15, 25, 50, and 100 μg/ml vs. tris-based diluent only as control. Samples were subjected to freezing then thawing to evaluate the vital sperm characteristics. It was discovered that kaempferol at a concentration of 25 and 50 μg/ml of significantly (p