Area of ram sperm head modified during the addition of cryprotectant agents

PELUFO V.; VENTURINO A; MALCOTTI V.; AISEN E

Madrid

Congreso; 55th Annual Meeting Society for Cryobiology CRYO2018; 2018

Society for Cryobiology

Sperm freezing procedures have considerable variations regarding the steps beingemployed for cooling, freezing, and addition of cryoprotectants. The area of the sperm head (ASH) is closely related to cell volume, so it was determined for the study of sperm cell volume changes during the dilution and semen cooling. In this work, the effects of the addition of glycerol and/or trehalose on ASH, either before (30°C) or after cooling (5°C), were evaluated. The different samples were observed at 30°C initial time (T0), at 5°C-90 min (T90) and at pre-freezing (Tpf). The ASH was determined using Rose Bengal dye and the NIS-element AR® program. At T0, not differences were observed, with an average value of 34.96 ± 0.39 μm2. At T90, all treatments significantly increased the ASH with respect to the control. At Tpf, all treatments showed an increase of ASH, similar to that registered at 90 min, except for glycerol 30°C + trehalose 5°C treatment, which reduced this parameter to values similar to the control. The analysis of the addition of cryoprotectants at Tpf with respect to those at T0 allows to affirm that the diluents containing trehalose reduce the flow of water into the sperm cells, while the diluent containing only glycerol, added at Tpf, showed a significant increase in ASH, and consequently an increase in cell volume (35.52 ± 0.33 μm2 at T90 vs. 36, 89 ± 0.32 μm2 at Tpf, p