• Pre-incubation with Kisspeptin improves the adverse effects of the freeze-thawed Human Ejaculated sperms
  • Seyede Fateme Hosseini,1,*
    1. Anatomy Department, Shiraz University of Medical Sciences, Shiraz, Iran


  • Introduction: Spermatozoa cryopreservation results in reduction of sperm quality. Kisspeptin (KP), as an antioxidant, has beneficial effects on the sperm functions. The objectives of this study were to determine the mitigating impact of KP on detrimental effects of the sperm freeze-thawing process and compare it with glutathione (GSH), as the positive control.
  • Methods: Thirty normal semen samples, prepared by swim-up procedure, were divided into three aliquots: negative control, without any treatment; positive control receiving GSH; and experimental aliquot treated with KP for 30 min. All aliquots were cryopreserved, and then thawed after 48hr. Sperm motility was assessed according to WHO guidelines. Acrosomal reaction and capacitation were evaluated by FITC-conjugated Peanut agglutinin (PNA), Wheat germ agglutinin (WGA), and Concanavalin A (ConA); the percentage of the positive cells was evaluated by flowcytometry. Sperm DNA quality was evaluated by Acridine Orange, Aniline Blue, Chromomycin A3, and TUNEL staining methods. Statistical analyses were performed using ANOVA and LSD.
  • Results: KP supplementation improved the motility and led to an increase in the percentage of capacitated and acrosome-intact sperms compared with both controls. Freeze-thaw procedure damaged DNA integrity severely, and KP pre-treatment significantly reduced the frequency of apoptotic sperms along with those with histone – protamine substitution impairment.
  • Conclusion: Pre-exposure of the sperms to KP can protect the sperm quality including motility and DNA integrity against the detrimental influence of freezing and thawing. Therefore, it can be considered as a good pre-additive substance to control the sperm quality during freezing and thawing procedure.
  • Keywords: Kisspeptin, motility, DNA integrity, Cryopreservation, Lectin histochemistry