Effects of cryoprotectants and trehalose on electron microscopic evaluation of cryopreserved sperm

Background: The present study was conducted to examine the effects of different cryoprotectants (glycerol, G and ethylene glycol, EG) and trehalose (T) on cryopreserved ram sperm.Methods: Ejaculates collected from six Merinos rams were pooled and evaluated at 37 C. Pooled semen samples were divided into six aliquots, and diluted in a Tris-based extenders containing %5 G, %3 G + 60 mM T, % 1.5 G + 100 mM T , % 5 EG, %3 EG + 60 mM T and % 1.5 EG + 100 mM T. Then, they were cooled to 5 C and frozen in 0.25 ml French straws. Frozen straws were then thawed individually at 37 C for 25 s in a water bath for electron microscopy evaluation. Field Emission Scanned Electron Microscope (FESEM) was used for examining the thawed sperm.Result: The groups of ethylene glycol % 5 and %1.5 ethylene glycol + 100 mM trehalose provided the highest protection for sperm morphologies among the groups. The addition of trehalose at high dose in semen extenders tended to reduce the damaged percentages in sperm cells (pConclusion: Trehalose at 100 mM with/without ethylene glycol (1.5 and 5%) supplementation in semen extenders provided a great protection of sperm morphologies against cryopreservation injury in electron microscopic examination.

کلیدواژه ها:

Cryopreservation ، Cryoprotectant ، Sperm parameters ، Trehalose ، Ram semen