Effect of methyl-beta-cyclodextrin on the viability and acrosome damage of sex-sorted sperm in frozen-thawed bovine semen

BACKGROUND: The regulation of methyl-beta-cyclodextrin (MBCD) on cryodamage on X- and Y-sperm during cryopreservation of semen was investigated. The semen was collected from ten healthy bulls of proven fertility by an artificial vagina. The bovine sperm treated with MBCD fresh solution (0, 1, 5, 10, and 20 mM). The sperms were evaluated for viability and acrosome damage using flow cytometry. Moreover, X- and Y-sperm in frozen-thawed bovine semen were sorted by flow cytometry after Hoechst 33342-dyed, and the viability and acrosome damage of sperms were analyzed. RESULTS: Sperm viability in frozen-thawed semen was decreased by MBCD (p < 0.05), also the acrosome damage of sperm was significantly increased (p < 0.05). Moreover, we sorted X- and Y-sperm from frozen-thawed bovine semen for observing the viability and acrosome damage on the separated X- and Y-sperm after MBCD treatment. Viability of X-sperm was significantly lower than that of Y-sperm (p < 0.05). Also, acrosome damage of X-sperm was significantly higher than Y-sperm (p < 0.05). CONCLUSIONS: Methyl-beta-cyclodextrin enhances damage of sperm in frozen-thawed bovine semen, and X-sperm is more sensitive than Y-sperm in cell damage. These results demonstrate that MBCD can inhibit viability of spermatozoa in frozen-thawed bovine semen (for X-sperm, especially).