Background: Sperm quality has an important role in the success of assisted reproductive techniques, by adding some bioactive agents with a positive impact on sperms, it can be improved. Objective: This study aimed to evaluate the effects of kisspeptin on the sperm motility criteria, lactate dehydrogenase-C (LDHC) activity, acrosomal reaction, and capacitation in the mouse testicular sperm in vitro. Materials and Methods: Sperm samples were extracted from testes of 96 male Balb/C mice weighing 25-30 gr, aged 6-8 wk. Then, they were separated into 4 parts; 2 controls and 2 kisspeptin-treated aliquots; each one incubated for either 15 or 30 min. The sperm motility and the LDHC activity were evaluated, and also the frequency of the non-capacitated, intact, and acrosomal-reacted sperms were evaluated by staining with Wheat germ agglutinin, Peanut agglutinin, and Concanavalin A, respectively. The stained sperms were analyzed by flow cytometry and fluorescent microscope. Results: Our result showed that kisspeptin increased both the sperm motility (p = 0. 04) and LDHC enzyme activity (p = 0. 04) after 15 min of incubation. At the same time, it did not impact the frequency of the non-capacitated, intact and acrosomal-reacted sperms after incubation in the same period (p = 0. 16). Conclusion: A 15 min period of incubation with kisspeptin could be applicable for evaluating sperm motility and LDH activity.