Cocchia N, Ciani F, El-Rass R, Russo M, Borzacchiello G, Esposito V, Montagnaro S, Avallone L, Tortora G, Lorizio R
Department of Veterinary Clinic Sciences, University of the Studies of Naples Federico II, Naples, Italy.
Zygote. 2010 Feb;18(1):1-8. doi: 10.1017/S0967199409990256. Epub 2009 Nov 26.
Cryopreservation of gametes is an important tool in assisted reproduction programmes; long-term storage of oocytes or spermatozoa is necessary when in vitro fertilization (IVF) or artificial insemination is to be performed at a future date. Cryopreservation of epididymal spermatozoa offers a potential tool for rescuing genetic material from males of endangered populations. The objectives of this work were to: (1) examine sperm motility, viability, abnormality and acrosome integrity of frozen-thawed domestic cat epididymal spermatozoa; and (2) evaluate the same cryopreservation method on wild feline spermatozoa, needed to preserve their genetic resources. Epididymides were collected from 20 domestic cats during routine neutering procedure and from two wild felines at autopsy. The sperm samples, diluted with 4% glycerol/Tris/egg yolk, were loaded into 0.25 ml mini-straws, exposed to nitrogen vapour and stored in liquid nitrogen. After 4 weeks, samples were thawed and re-evaluated. The quality of each fresh and frozen-thawed sperm sample was tested by determining the motility (54.7 +/- 11.3% and 32 +/- 13.1% respectively for cat spermatozoa; 38.3 +/- 18.7% and 21.5 +/- 16.8% respectively for tiger spermatozoa), viability (74.3 +/- 8.6% and 45.2 +/- 9.4% respectively for cat spermatozoa; 42.4 +/- 14.5% and 33.5 +/- 12.9% respectively for wild felid spermatozoa), morphology and acrosomal status. The present study showed that feline epididymal spermatozoa can be frozen in egg-yolk extender with 4.0% glycerol in 0.25 ml straws. The procedure used in the present study for epididymal cat sperm cryopreservation may be applied to bank the genetic resources of wild felid species.
配子的冷冻保存是辅助生殖计划中的一项重要工具；当计划在未来某个时间进行体外受精（IVF）或人工授精时，卵母细胞或精子的长期储存是必要的。附睾精子的冷冻保存为拯救濒危种群雄性个体的遗传物质提供了一种潜在工具。本研究的目的是：（1）检测冻融后的家猫附睾精子的活力、生存力、异常情况和顶体完整性；（2）评估同一冷冻保存方法对野生猫科动物精子的效果，这对于保护它们的遗传资源是必要的。在常规绝育手术期间从20只家猫以及在尸检时从两只野生猫科动物身上采集附睾。将用4%甘油/Tris/蛋黄稀释的精子样本装入0.25 ml的微型细管中，置于氮蒸气中并储存在液氮中。4周后，将样本解冻并重新评估。通过测定活力（家猫精子分别为54.7±11.3%和32±13.1%；虎精子分别为38.3±18.7%和21.5±16.8%）、生存力（家猫精子分别为74.3±8.6%和45.2±9.4%；野生猫科动物精子分别为42.4±14.5%和33.5±12.9%）、形态和顶体状态来检测每个新鲜和冻融后精子样本的质量。本研究表明猫附睾精子可以在含有4.0%甘油的蛋黄稀释液中于0.25 ml细管中冷冻。本研究中用于猫附睾精子冷冻保存的方法可应用于储存野生猫科动物物种的遗传资源。
