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首页> 外文期刊>Cryobiology: International Journal of Low Temperature Biology and Medicine >Improvement of European eel sperm cryopreservation method by preventing spermatozoa movement activation caused by cryoprotectants.
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Improvement of European eel sperm cryopreservation method by preventing spermatozoa movement activation caused by cryoprotectants.

机译：通过防止冷冻保护剂引起的精子运动活化，改进欧洲鳗精子冷冻保存方法。

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摘要

Sperm production has been obtained from European and Japanese eels, but its quality and quantity tend to be changeable. So, its cryopreservation has been tried in both species. Dimethyl sulfoxide (Me(2)SO) is the best cryoprotectant for European eel sperm, but increases the medium osmolality, inducing the activation of spermatozoa motility. To avoid this, different combinations of pH (6.5 and 8.5) and NaHCO(3) concentrations (20, 40 and 80mM) were tested with two Me(2)SO concentrations (5% and 10%). Foetal bovine serum (FBS, 25%v/v) was added as a membrane protector to all the freezing media used in the different experiments. The highest Me(2)SO and NaHCO(3) concentrations at pH 6.5 caused the best post-thawing motility (26+/-4%). A second experiment was carried out testing media with Me(2)SO 10% with additional NaHCO(3) concentrations (100 and 120 mM). The highest post-thawing motility (38+/-3%) was found in the media containing NaHCO(3) 100mM, but no significant difference was observed compared with the best in the previous experiment (NaHCO(3) 80 mM). In a parallel experiment, aiming to improve the protection against the cryopreservation process, bovine serum albumin (BSA, 5%w/v) was added instead of FBS. Lower motilities were registered with BSA as membrane protector. Spermatozoa activation caused by addition of Me(2)SO can be prevented using high NaHCO(3) concentrations, improving the cryopreservation process. This effect seems be based on some of the products dissociated from NaHCO(3) in aqueous solution, affecting the intracellular pH, essential in the sperm motility.
机译：精子的生产是从欧洲和日本的鳗鱼那里获得的，但是其质量和数量往往会发生变化。因此，已经在两种物种中尝试了冷冻保存。二甲基亚砜（Me（2）SO）是欧洲鳗精子的最佳防冻剂，但会增加中等渗透压，诱导精子活力的激活。为避免这种情况，使用两个Me（2）SO浓度（5％和10％）测试了pH（6.5和8.5）和NaHCO（3）浓度（20、40和80mM）的不同组合。将胎牛血清（FBS，25％v / v）作为膜保护剂添加到不同实验中使用的所有冷冻培养基中。在pH 6.5时，最高的Me（2）SO和NaHCO（3）浓度导致最佳的解冻后运动性（26 +/- 4％）。进行了第二项实验，测试了含10％Me（2）SO和其他NaHCO（3）浓度（100和120 mM）的介质。在含有100mM NaHCO（3）的培养基中发现最高的解冻后运动性（38 +/- 3％），但与先前实验中的最佳（NaHCO（3）80 mM）相比，没有观察到显着差异。在平行实验中，为了提高对冷冻保存过程的保护作用，添加了牛血清白蛋白（BSA，5％w / v）代替FBS。向BSA注册了较低的功能作为膜保护剂。可以使用高浓度的NaHCO（3）来防止由于添加Me（2）SO引起的精子活化，从而改善冷冻保存过程。这种作用似乎是基于从水溶液中的NaHCO（3）分离出来的某些产物，影响了精子运动所必需的细胞内pH。

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