Effects of different cryoprotectants on the cryopreservation of cattle testicular tissue

Abstract. Cryopreservation of testicular tissue is a new option in fertility preservation for prepubertal male animals. The purpose of this study was to explore the effects of different cryoprotectant agents (CPAs) at various concentrations on testes after the cryopreservation of calf testicular tissue. These experiments selected dimethyl sulfoxide (DMSO), glycerol, propylene glycol (PrOH), and sucrose as CPAs in varying doses (2.5, 5, 7.5, 10, 12.5, 15, 17.5, and 20 %; v/v) in 8-month-old calf testicular tissue that was frozen and preserved. Then, cell viability, testosterone production, malondialdehyde (MDA) level, and superoxide dismutase (SOD) level were detected and analyzed following cryopreservation. The results showed that the optimal concentrations of DMSO, PrOH, glycerol, and sucrose were 10, 10, 7.5, and 10 %, respectively. Compared to the optimal concentrations of CPAs, cell viability and testosterone production decreased significantly at a lower and higher CPA concentration (P < 0.05). At the optimal concentrations of CPAs, the DMSO group showed higher cell viability and testosterone production than other CPA groups (P < 0.05). Compared to the optimal concentration of CPAs, the MDA level increased and the SOD level decreased at a lower or higher concentration of CPAs, but there was no significant difference (P > 0.05). Cell viability was significantly positively correlated with testosterone production (P < 0.05). In conclusion, DMSO provided the most effective protection for calf testicular tissue cryopreservation and the optimal concentration was 10 %.