Laboratory of Animal Germplasm Conservation, Federal Rural University of Semi-Arid, Mossoro-RN, Brazil.
Laboratory of Animal Biotechnology, Federal Rural University of Semi-Arid, Mossoro-RN, Brazil.
Cryobiology. 2021 Apr;99:40-45. doi: 10.1016/j.cryobiol.2021.01.016. Epub 2021 Jan 27.
Addressing the establishment of biobanks for the conservation of wild hystricomorph rodents' germplasm, we verified the effects of different extenders and distinct concentrations of non-permeant cryoprotectants on the sperm parameters of Spix's yellow-toothed cavies. Nine testis-epididymis complexes were used for sperm collection by retrograde washing using Tris or a powdered coconut water extender (ACP®-116c). Spermatozoa were diluted and frozen with the same extenders supplemented with egg yolk or Aloe vera at a 10% or 20% concentration. After recovery and cryopreservation, all samples were evaluated for sperm kinetic parameters, morphology, membrane integrity, osmotic response, and sperm-binding capability using an egg yolk perivitelline membrane assay. After recovery, no differences were observed between Tris and ACP®-116c that provided 515.4 × 10 sperm/mL and 561.6 × 10 sperm/mL, presenting >65% motile sperm, respectively. After cryopreservation, most effective preservation of sperm kinetic parameters (68.1 ± 5.9% motile sperm) and membrane integrity (48.2 ± 7.4%) was provided by Tris extender supplemented with 10% egg yolk. However, both extenders supplemented with any concentration of egg yolk or Aloe vera presented similar preservation of osmotic response and sperm-binding ability after cryopreservation. In summary, we suggest the use of a Tris extender supplemented of 10% egg yolk for cryopreservation of Spix's yellow-toothed cavy epidydimal sperm.
为了建立用于保护野生豪猪目啮齿动物种质资源的生物银行，我们验证了不同稀释液和不同浓度的非渗透性冷冻保护剂对黄牙豚鼠精子参数的影响。通过使用 Tris 或粉状椰子水稀释剂（ACP®-116c）进行逆行冲洗，从 9 个睾丸-附睪复合体中收集精子。将精子稀释并用相同的稀释液冷冻，其中添加了 10%或 20%浓度的蛋黄或芦荟。恢复和冷冻保存后，使用蛋黄卵黄膜试验评估所有样品的精子动力学参数、形态、膜完整性、渗透反应和精子结合能力。恢复后，Tris 和 ACP®-116c 之间没有差异，它们分别提供了 515.4×10 个/mL 和 561.6×10 个/mL 的精子，分别有 >65%的运动精子。冷冻保存后，Tris 稀释液中添加 10%蛋黄提供了最有效的精子动力学参数（68.1±5.9%的运动精子）和膜完整性（48.2±7.4%）保存。然而，添加任何浓度的蛋黄或芦荟的两种稀释液在冷冻保存后都具有相似的渗透反应和精子结合能力保存。总之，我们建议使用 Tris 稀释液补充 10%的蛋黄来冷冻保存黄牙豚鼠的附睪精子。
