Yasui George Shigueki, Arias-Rodriguez Lenin, Fujimoto Takafumi, Arai Katsutoshi
Graduate School of Fisheries Science, Division of Marine Life Sciences, Laboratory of Aquaculture Genetics and Genomics, Hokkaido University, Hokkaido, Japan.
Cryo Letters. 2008 Sep-Oct;29(5):383-90.
Here, we propose a simple and inexpensive method for fish sperm cryopreservation. Sperm samples of the loach Misgurnus anguillicaudatus (Teleostei: Cobitidae) were diluted 7-fold by an extender containing 63.5 mM NaCl, 114 mM KCl, 20 mM Tris and 10% methanol. The cryogenic straws were placed in three kinds of self-made tubes which diameter was changed by commercially available materials and then immersed into powdered dry ice for 2 min and plunged into liquid nitrogen. This procedure resulted in a cooling rate at -421.4 +/- 119.84 (control), -55.8 +/- 4.32 (tube 1), -40.2 +/- 3.43 (tube 2) and -33.3 +/- 2.09 C/min (tube 3). In the slowest cooling rate by the tube 3, total motility (72 +/- 3 %), duration (146 +/- 12 s) and hatching rates (29 +/- 04 %) were higher than those by other rates. Progressive motility (83 +/- 5 %) did not differ significantly from fresh samples.
在此，我们提出一种简单且经济的鱼类精子冷冻保存方法。将泥鳅（硬骨鱼纲：鳅科）的精子样本用含有63.5 mM氯化钠、114 mM氯化钾、20 mM Tris和10%甲醇的稀释液稀释7倍。将冷冻细管置于三种通过市售材料改变直径的自制管中，然后浸入干冰粉末中2分钟，再投入液氮中。该程序导致的冷却速率分别为-421.4 +/- 119.84（对照）、-55.8 +/- 4.32（管1）、-40.2 +/- 3.43（管2）和-33.3 +/- 2.09℃/分钟（管3）。在管3导致的最慢冷却速率下，总活力（72 +/- 3%）、持续时间（146 +/- 12秒）和孵化率（29 +/- 4%）高于其他速率下的数值。前进活力（83 +/- 5%）与新鲜样本相比无显著差异。
