Legendre M, Billard R
Reprod Nutr Dev (1980). 1980;20(6):1859-68. doi: 10.1051/rnd:19801012.
Cryopreservation trials on rainbow trout (Salmo gairdneri) sperm were carried out using two basic extenders: Mounib's medium (M) and Ménézo's medium (Me) to which were added bovine serum albumin (BSA) and tellurite egg yolk (Institut Pasteur). After 10 p. 100 of DMSO was added to these different deep-freeze diluents (DD), they were mixed with the sperm and then deep-frozen into 100 microliter pellets on dry ice. The pellets were stored in liquid nitrogen for periods lasting from 3 days to 6 months. The intensity of sperm motility and fertilizing ability were measured before and after cryopreservation. After the sperm was diluted in Ménézo's medium, slight spermatozoon motility was noticed, which probably caused their early exhaustion and would explain the lower fertilizing ability observed after thawing. Mounib's medium gave better results, especially after 10 p. 100 of egg yolk was added. The optimal deep-freeze conditions were: 1/3 dilution, no equilibration after dilution but immediate deep-freezing at a rate of 10 to 40 degrees C/min. Thawing had to be carried out rapidly in 10 sec. However, the spermatozoa were altered during the freezing-thawing process, and during insemination more frozen spermatozoa had to be used to equal the fertilization rate obtained with non-frozen sperm. However, the fertile spermatozoa gave normal embryogenesis and no abnormal development was seen up to the vesicle resorption stage. At the end of spermiation, sperm fitness for deep-freezing decreased, perhaps due to sperm senescence. Pooling the sperm of several males partially compensated for the loss of fertilizing ability seen at the end of the reproductive period.
使用两种基本的稀释液对虹鳟（Salmo gairdneri）精子进行冷冻保存试验：穆尼布培养基（M）和梅内佐培养基（Me），并向其中添加牛血清白蛋白（BSA）和亚碲酸盐蛋黄（巴斯德研究所）。在这些不同的深度冷冻稀释液（DD）中加入10%（体积分数）的二甲亚砜（DMSO）后，将它们与精子混合，然后在干冰上冷冻成100微升的小丸。这些小丸在液氮中保存3天至6个月。在冷冻保存前后测量精子活力强度和受精能力。精子在梅内佐培养基中稀释后，观察到精子有轻微活力，这可能导致它们过早耗尽，这也可以解释解冻后观察到的较低受精能力。穆尼布培养基的效果更好，尤其是添加10%（体积分数）蛋黄后。最佳深度冷冻条件为：1/3稀释，稀释后不进行平衡处理，但立即以10至40℃/分钟的速率深度冷冻。解冻必须在10秒内快速进行。然而，精子在冻融过程中发生了变化，并且在授精过程中必须使用更多的冷冻精子才能达到与未冷冻精子相同的受精率。然而，有活力的精子产生了正常的胚胎发育，直到囊泡吸收阶段都没有观察到异常发育。在精子排放末期，精子的冷冻适应性下降，这可能是由于精子衰老所致。将几只雄性的精子混合部分弥补了繁殖期结束时观察到的受精能力损失。
