Cryopreservation of rainbow trout sperm in large volume straws:: application to large scale fertilization
Cryopreservation of rainbow trout sperm in large volume straws:: application to large scale fertilization
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DOI：
10.1016/s0044-8486(01)00636-6
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发表时间：
2001-10-01
期刊：
影响因子：
4.5
通讯作者：
Herráez, MP
中科院分区：
文献类型：
--
作者：
Cabrita, E;Robles, V;Herráez, MP
传统的0.25毫升和0.5毫升吸管已成功地应用于虹鳟鱼精子的超低温保存。为了便于大批量受精，并将这项技术推广到商业孵化场，研究了1.8毫升和5毫升吸管的适宜性。精子在#6 Erdahl和Graham稀释液中以1：3的比例稀释，其中添加7%DMSO、10%蛋黄和7.5 mg/mlPromine。用泡沫塑料在液氮表面以上的不同高度进行冷冻，并在不同温度的水浴中解冻。用热电偶记录吸管内的冻结和解冻速度。精子冷冻保存的成功是通过几个精子质量参数来检验的：活动率、存活率和膜功能。生育能力测试也是小批量进行的，使用200个鸡蛋/毫升稀释精子的比率。1.8毫升和0.5毫升吸管的冷冻和解冻速度相似。然而，即使在较低的冻结和较高的解冻温度下，使用5毫升的吸管也可以实现较低的冻结和解冻速度。精子活动率分析在所有受试吸管(0.5、1.8和5ml)中冷冻保存的精子中得到了类似的结果。然而，大容量培养的细胞存活率(61.9%)和受精率(73.2%)略低于0.5ml培养管(分别为77.4%和84%)。用冷冻在5毫升吸管中的精子进行大批量卵子(1600-2000年)的受精，提供了相似的受精率(73%)，这表明5毫升吸管的好处弥补了受精率的轻微损失。我们的结果表明，大量精子的冷冻保存可以用于孵化目的。(C)2001 Elsevier Science B.V.保留所有权利。
Traditional 0.25- and 0.5-ml straws have been successfully applied to the cryopreservation of rainbow trout sperm. In order to facilitate the fertilization of large egg batches, and with the purpose of transferring this technique to commercial hatcheries, the suitability of 1.8 and 5 ml straws was studied. Sperm was diluted at a 1:3 ratio in #6 Erdahl and Graham extender with 7% DMSO, 10% egg yolk and 7.5 mg/ml Promine. Freezing was performed at different levels above the surface of liquid nitrogen using a styrofoam box and thawing was carried out in a water bath at different temperatures. Freezing and thawing rates inside the straws were registered with a thermocouple. The success of sperm cryopreservation was checked in vitro using several sperm quality parameters: motility, viability and membrane functionality. Fertility tests were also performed on small batches using a ratio of 200 eggs/ml of diluted sperm. Freezing and thawing rates were similar in 1.8- and 0.5-ml straws. However, lower freezing and thawing rates were achieved with 5-ml straws even at lower freezing and higher thawing temperatures. Motility analysis gave similar results in sperm cryopreserved in all the tested straws (0.5, 1.8 and 5 ml). However, cell viability (61.9%) and fertility (73.2%) were slightly lower in large volumes than in 0.5-ml straws (77.4% and 84%, respectively). Fertilization of larger egg batches (1600-2000 eggs) with sperm frozen in 5-ml straws provided similar fertilisation rates (73%), demonstrating that the slight loss in fertility is compensated by the benefits of 5-ml straws. Our results showed that cryopreservation of large sperm volumes could be useful for hatchery purposes. (C) 2001 Elsevier Science B.V. All rights reserved.