四种渗透性防冻剂在猕猴精子低温冷冻保存中对精子功能状态的影响

以冷冻精子的复苏运动度、荧光染料Hoechst 3 3 2 5 8检测的细胞膜完整率、异硫氰酸荧光素标记的花生凝集素 (FITC PNA)检测的顶体完整率作为精子功能状态的指标 ,对甘油、二甲亚砜、乙二醇和丙二醇 4种常用渗透性防冻剂在猕猴精子冷冻保存过程中的作用进行了比较。结果表明 :冷冻保存精子的复苏运动度 ,甘油 ( 4 7 3± 5 7% )和乙二醇 ( 4 4 8± 6 7% ) >二甲亚砜 ( 2 2 9± 0 9% ) >丙二醇 ( 0± 0 % ) ;细胞膜完整率 ,甘油 ( 5 4 8± 3 2 % )和乙二醇 ( 5 4 0± 6 7% ) >二甲亚砜 ( 3 7 5± 7 0 % ) >丙二醇 ( 2 8 3± 6 5 % ) ;顶体完整率 ,甘油 ( 82 2± 2 4 % )和乙二醇 ( 82 4± 2 4 % ) >二甲亚砜 ( 6 8 7± 5 7% )和丙二醇 ( 72 3±3 5 % ) (P <0 0 5 )。结果提示 :二甲亚砜和丙二醇 ,尤其是丙二醇并不适合猕猴精子的冷冻保存 ;而乙二醇具有和甘油相似的保护作用 ,是一种极具潜力的猕猴精子冷冻保存的渗透性防冻剂。

Cryoprotective Effect of Four Penetrating Cryopro tectants on Rhesus Monkey (Macaca mulatta) Spermatozoa Cryopreservation

The present study aimed to examine four cryoprotectants:glycerol,dimethyl sulfoxide,ethylene glycol and propylene glycol on rhesus monkey sperm cryopreservation.Frozen-thawed sperm cryo-survival and function was evaluated by sperm motility,plasma membrane integrity and acrosome integrity,and sperm plasma membrane and acrosome status was determined by nuclei stain Hoechst 33258 and Fluorescein isothiocyanate-conjugated peanut agglutinin (FITC-PNA).The result showed that:for the ability of protection on post-thaw sperm motility,glycerol (47.3±5.7%) and ethylene glycol (44.8±6.7%)> dimethyl sulfoxide (22.9±0.9%)> ethylene glycol (0±0%);for the ability of protection on sperm plasma membrane integrity,glycerol (54.8±3.2%) and ethylene glycol (54.0±6.7%)> dimethyl sulfoxide (37.5±7.0%)> ethylene glycol (28.3±6.5%);for the ability of protection on sperm acrosome integrity,glycerol (82.2±2.4%) and ethylene glycol (82.4±2.4%)> dimethyl sulfoxide (68.7±5.7%) and ethylene glycol (72.3±3.5%) (P<0.05).The results indicate that cryoprotective effects of various penetrating cryoprotectants are different.Dimethyl sulfoxide and propylene glycol are not suitable for rhesus monkey sperm freezing,especially propylene glycol.Ethylene glycol with similar cryoprotective properties to glycerol could be successfully used in the cryopreservation of rhesus monkey spermatozoa.