文章摘要
王梅芳.马氏珠母贝精子的超低温保存[J].水产学报,2006,30(2):
马氏珠母贝精子的超低温保存
Study on cryopreservation of spermatozoa in Pinctada martensii
投稿时间:2014-03-08  修订日期:2014-03-08
DOI:
中文关键词: 马氏珠母贝 精子 超低温保存
英文关键词: Pinetada martensii spermatozoa cryopreservation
基金项目:国家海洋“863”项目(2002AA603022);广东省科技计划项目(2003C20330)
作者单位地址
王梅芳 广东海洋大学水产学院 广东海洋大学水产学院
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中文摘要:
      通过比较不同pH值(5.5-9.5)、不同盐度的海水等作为基础液对马氏珠母贝精子保存的影响,选择其中无激活精子作用又对其生理特性(活力,寿命,受精能力等)无影响者,加入二甲亚砜抗冻剂配制成超低温保存的抗冻保护液,精液与保护液按1:10和1:20的比例混合,样品按4组不同的降温程序平衡后转入液氮冷冻,比较其超低温冻存的效果。结果表明:以海水(pH7.5—8.0)为基础液,配制10%DMSO作为抗冻保护液,精液与保护液比例为1:20,在低温(0-4℃)中平衡约30min,在距液氮面15cm、5cm处分别停留5min、10min后转入液氮(-196℃),冻存精子效果良好。冷冻24h、48h及5个月后,在38—40℃下水浴解冻复苏后,用终浓度0.25‰的氨海水刺激,精子存活率可超过60%,受精率可达80%。
英文摘要:
      Comparing solutions of different pH (5.5 - 9.5) and different salinity to develop suitable basic solution for spermatozoa cryopreservation of pearl oyster Pinctada martensii, we chose the basic solution, which could inhibit the activation and had less influence on the physiological characteristics (such as activity, life span and fertilizing capability) of the spermatozoa, and adopted dimethyl sulfoxide (DMSO) as antifreeze to compound diluents for protecting the spermatozoa. After combination of natural seawater with DMSO (10%) and seminal fluid at the ratios of 1 : 10 and 1:20, the specimens were transferred into LN2 by 4 temperaturelowering processes. The conclusion showed that cryopreservation processes were effective in which the samples were pre-frozen in 0- 8 ℃ within 30 minutes, swung at 15 cm above for 5 min and then at 5 cm above LN2 for 10min, transferred them into LN2 for prolonging period (24 h, 48 h and 5 months ), and then thawed them with water at 35-40 ℃, the survival rate of spermatozoa could exceed 60% when activated with 0.25‰ ammonia seawater and the fertilization rate could reach 80%.
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