不同冷冻方法和解冻液对猪精液冷冻效率的影响  

Effects of Different Freezing Methods and Thawing Solutions on Boar Semen Cryopreservation Efficiency

作　　者：周鑫[1] 张曼玲[1] 赵丽华[2] 李荣凤[1,2] 

机构地区：[1]内蒙古大学哺乳动物生殖生物学及生物技术教育部重点实验室,呼和浩特010021 [2]南京医科大学代谢疾病研究中心,南京210029

出　　处：《内蒙古大学学报（自然科学版）》2013年第1期80-86,共7页Journal of Inner Mongolia University：Natural Science Edition

基　　金：国家自然科学基金资助项目(30871408);国家高技术研究发展计划(863计划)(2009AA10Z111)

摘　　要：采用三种不同的冷冻方法冷冻猪的新鲜精液,再分别用不同的解冻液在相同条件下同时解冻,对比解冻后精子的活力和顶体完整率,以优化猪精液冷冻方法.实验一:随机选取三头健康的有稳定受精能力的优良种公猪,采集新鲜精液,分别用干冰颗粒、液氮颗粒和液氮细管三种方法冷冻,用同样的方法解冻,对比解冻后精子的活力和顶体完整率;实验二:用不同的解冻液解冻经干冰颗粒法冷冻的精液,对比不同解冻液对精子活力的影响;实验三:以孤雌激活和鲜精受精为对照,验证经实验一和实验二选择出来的最优冷冻方法和解冻液处理后的精子的体外受精能力.实验一显示,干冰颗粒冷冻法冷冻的猪精液解冻后精子活力(0.23)略高于液氮颗粒法的精子活力(0.20),而二者均显著高于细管冻精法的精子活力(0.08)(P<0.05),液氮颗粒法的顶体完整率(54.7%)和细管法的顶体完整率(54.2%)无显著差异,但二者均显著低于干冰颗粒法的顶体完整率(58.0%)(P<0.05).实验二显示,Glucose-EDTA解冻液解冻后,精子活力(0.22)显著高于DPBS-BSA解冻液解冻后的精子活力(0.13)(P<0.05).实验三:经干冰颗粒法冷冻、Glucose-EDTA解冻液解冻的精液,体外受精后的囊胚发育率(17.6%)与鲜精体外受精囊胚率(17.9%)无显著差异,但二者均显著低于孤雌激活对照组的囊胚率(36.6%)(P<0.05).由此可知:干冰颗粒法冷冻结合Glucose-EDTA解冻液解冻是较为理想猪精液冷冻保存方法,经该方法冷冻、解冻的猪精液,体外受精后的胚胎发育率与鲜精没有显著差异.Three different methods were applied to freeze boar fresh semen and two kinds of tha wing solution were used to thaw the frozen sperm. The optimal freezing method and thawing solu tion for boar semen eryopreservation were identified based on the motility of thawed sperm and the percentage of sperm with integrity acrosome. In experiment 1：Three health and fertile boars were used for semen collection. The fresh semen was frozen by three different methods（dry ice/pellet, liq uid nitrogen/pellet and liquid nitrogen/straw）. The sperm motility and the percentage of sperm with integrity acrosome were compared after thawing in same way. In experiment 2：The frozen sperm was thawed with different thawing solutions and the sperm motility and acrosome integrity were compared. In experiment 3 ： Based on experiment 1 and 2, the sperms frozenthawed by the best freezing method and thawing solution were selected for in vitro fertilization experiment. The devel opment of IVF embryos derived from frozen sperm were investigated with IVF embryos derived from fresh sperm and parthenogenetic embryos as control. Results were as following ： 1, The motility of sperm frozen by dry ice/pellet（0.23） was higher than that by liquid nitrogen/pellet（0.20） ,after thawing. Both of them were significantly higher than that（0.08）by liquid nitrogen/Straw（P〈0. 05）. The percentage of sperm with integrity acrosome in dry ice/pellet group （58.0%） was signifi cantly different from that in liquid nitrogen/pellet group （54. 7%） and liquid nitrogen nitrogen/ straw group（54.2%）,but there were not significant difference between liquid nitrogen group and liquid nitrogen/straw group（P〈0.05）. 2, The motility of sperm thawed by a high glucoseEDTA solution （0.22） was significantly higher than that by a PBSBSA solution （0.13） （P〈 0.05）. 3, The sperm frozen by dry ice/pellet method and thawed in high glucoseEDTA were used for IVF. The results indicated that there was no significant difference in blastocyst

关 键 词：猪精液 冷冻方法 解冻液 效率 囊胚率 

分 类 号：S814[农业科学—畜牧学]