抗冻剂对水牛卵母细胞冷冻后孤雌发育的影响  

Effects of Antifreeze on Parthenogenetic Development of Buffalo Vitrified Oocytes

作　　者：杨素芳[1] 陈玉[1] 冯贵雪[2] 王晓丽[3] 陈自洪[1] 石德顺[1] 

机构地区：[1]广西大学动物繁殖研究所,南宁530005 [2]广西壮族自治区妇幼保健院生殖中心,南宁530003 [3]广西大学动物科学技术学院,南宁530005

出　　处：《西南大学学报（自然科学版）》2009年第12期8-13,共6页Journal of Southwest University(Natural Science Edition)

基　　金：国家863计划资助项目(2005AA206130);广西青年基金资助项目(桂科青0542022)

摘　　要：试验主要探索了不同抗冻剂组合对水牛MⅡ期卵母细胞冷冻-解冻后孤雌发育潜力的影响.通过对3种抗冻剂组合(Ⅰ:20%乙二醇(EG)+20%二甲基亚砜(DMSO);Ⅱ:20%EG+20%1,2-丙二醇(PROH);Ⅲ:20%EG+20%PROH+10%DMSO)的毒性试验,发现试验组的形态正常率均低于对照组(p<0.05),但Ⅰ组的分裂率、8-细胞率和囊胚率与对照组无显著差异(p>0.05).并以抗冻剂Ⅰ组做冷冻液,探讨了解冻液中蔗糖梯度浓度(A:0.5,0.25,0.10mol/L与B:0.62,0.42,0.31mol/L)对卵母细胞冷冻-解冻效果的影响.B组的形态正常率显著高于A组(p<0.05),且B组的分裂率和囊胚率与对照组无显著差异(p>0.05).最后比较了3种抗冻剂组合对卵母细胞冷冻-解冻后的孤雌激活发育效果.3组的形态正常率、分裂率和8-细胞率之间无显著差异(p>0.05),但均显著低于对照组(p<0.05),其中Ⅰ组的囊胚率与Ⅱ、Ⅲ组及对照组差异均不显著(p>0.05),Ⅱ组和Ⅲ组的囊胚率显著低于对照组(p<0.05).结果表明:采用抗冻剂组合Ⅰ冷冻水牛MⅡ期卵母细胞,解冻时采用B组解冻液去除抗冻剂,可以取得较好的冷冻保存效果.The aim of this study was to investigate the effects of different antifreeze combinations on parthenogenetic development of buffalo metaphase II （MII） vitrified-thawed oocytes.Toxicity test of 3 antifreeze combinations （I：20% ethylene glycol （EG）＋20% dimethyl sulfoxide （DMSO）;II：20% EG＋20% 1,2-propanediol （PROH）;and III：20% EG＋20% PROH＋10% DMSO） showed that the morphological normal rate in the 3 groups was all lower than that in the control group （p0.05）,but the cleavage rate,8-cell rate and blastocyst rate in Group I did not differ significantly from those of the control（p0.05）.Therefore,the thawing effect of sucrose concentration gradient in the thawing solution （A：0.5,0.25,0.10 mol/L;B：0.62,0.42,0.31 mol/L） on vitrification of oocytes was compared in Group I.The morphology normal rate of thawed oocytes in Group A was lowered than in Group B （p0.05）.In PA test,the cleavage rate and blastocyst rate of the thawed oocytes in Group B were shown to be similar to those of the control （p0.05）.Finally,the effects of the antifreeze combinations on parthenogenetic activation development of vitrified-thawed oocytes were compared.No significant differences in morphology normal rate,cleavage rate and 8-cell rate were noticed in the three groups （p0.05）,but they were all lower than those in the control （p0.05）.Blastocyst development rate in group I had no difference from Groups II and III and the control （p0.05）.However,blastocyst rate in group II and III was significantly lower than that in the control （p0.05）.In conclusion,a satisfactory antifreeze effect could be obtained when the buffalo MII oocytes were vitrified by group I and thawed by group B.

关 键 词：水牛 卵母细胞 玻璃化冷冻 抗冻剂 孤雌激活 

分 类 号：S823.83[农业科学—畜牧学]