title: Cryopreservation of stallion spermatozoa using different cryoprotectants and combinations of cryoprotectants.
abstract: The present study investigates the effects of five cryoprotectants (CPAs) and cryoprotectant combinations on the post-thaw total motility, progressive motility, viability, mitochondrial membrane potential and acrosome integrity in stallion spermatozoa. In Experiment 1, the objective was to compare the impact of different concentrations (2.5%, 3.5% and 5%) of a single CPA, including glycerol (Gly), ethylene glycol (EG), dimethyl sulphoxide (DMSO), methyl formamide (MF), and dimethylformamide (DMF) for stallion spermatozoa cryopreservation. In Experiment 2, two or more CPAs were used to assess whether this improved post-thaw spermatozoa quality. Gly, MF and DMF, were used to prepare seven combinations of freezing extender with different mixtures of cryoprotectant, and the 3.5% Gly, MF and DMF were used as a control group. The results show that post-thaw total motility, progressive motility, viability, and mitochondrial membrane potential for all concentrations of EG and DMSO were less than the 3.5% and 5% Gly and MF and 2.5% and 3.5% DMF (P<0.05). Use of the 3.5% concentration resulted in the greater post-thaw total motility and progressive motility than the 2.5% and 5% concentrations for all CPAs. The results for the use of different combinations of cryoprotectant indicate there are differences in progressive motility and viability. The viability with the use of Gly(2/3)+MF(1/3) was 44.65% and was greater than the Gly(1/3)+MF(1/3)+DMF(1/3) (30.96%), MF(2/3)+DMF(1/3) (35.05%), Gly (32.21%) and MF(33.76%) (P<0.05). The progressive motility with the use of the MF(2/3)+Gly(1/3) combination was 36.0% and was greater than in the DMF (25.0%) and MF(2/3)+DMF(1/3) (22.7%) (P<0.05). These results suggest that using the appropriate cryoprotectant combination instead of a single cryoprotectant can improve horse spermatozoa cryopreservation.
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi 832000, Xinjiang, China.
authors.forename: Zhuangyuan
authors.initials: Z
authors.lastname: Wu
authors.affiliation: Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China.
authors.forename: Xinbiao
authors.initials: X
authors.lastname: Zheng
authors.affiliation: Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China.
authors.forename: Yongming
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Huo
authors.affiliation: Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Dong
authors.affiliation: Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China.
authors.forename: Guoting
authors.initials: G
authors.lastname: Zhang
authors.affiliation: The Breeding Center of Yili Horse, Zhaosu 835600, Xingjiang, China.
authors.forename: Weihao
authors.initials: W
authors.lastname: Yu
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi 832000, Xinjiang, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Tian
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi 832000, Xinjiang, China.
authors.forename: Liangjun
authors.initials: L
authors.lastname: He
authors.affiliation: College of Animal Science and Technology, Shihezi University, Shihezi 832000, Xinjiang, China; Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi 830000, Xinjiang, China. Electronic address: chenjb126@126.com.
authors.forename: Jingbo
authors.initials: J
authors.lastname: Chen
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000008:administration & dosage / Q000494:pharmacology*; D006736:Horses / Q000502:physiology*; D008297:Male; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013094:Spermatozoa / Q000502:physiology*
chemical_list: D003451:Cryoprotective Agents
keywords: CPA; Cryoprotective agents; Equine spermatozoa; Frozen semen; Horse