脱防冻剂方法对牛体外受精胚胎冷冻后成活率的影响  

Effect of Dilution of Cryoprotectants on Survival Rate of Frozen Thawed Bovine IVF Embryos

作　　者：阳年生[1] 蒋和生[1] 王武陵[1] 石德顺[1] 卢克焕[1] 

机构地区：[1]广西农业大学畜牧兽医系

出　　处：《中国兽医学报》1998年第3期291-294,共4页Chinese Journal of Veterinary Science

基　　金：国家"863"课题

摘　　要：为了观察脱防冻剂方法对牛体外受精冷冻胚胎在体内、外发育率的影响，应用常规冷冻法在０．７５ｍｏｌ／Ｌ甘油＋０．５ｍｏｌ／Ｌ丙二醇溶液中冷冻受精后７、８ｄ的囊胚，解冻后在０．２５、０．５ｍｏｌ／Ｌ蔗糖液中二步或在０．２５ｍｏｌ／Ｌ蔗糖液中三步脱防冻剂的胚胎孵化率（分别为６８．６％、６２．２％、６８．７％）与用ＰＢＳ／ＦＣＳ六步脱防冻剂的差异不显著（７０．４％，Ｐ＞０．０５），但在０．５ｍｏｌ／Ｌ蔗糖液中三步脱防冻剂后，孵化率显著降低（４７．６％，Ｐ＜０．０１）。用０．２５ｍｏｌ／Ｌ或０．５ｍｏｌ／Ｌ蔗糖或海藻糖预先使胚胎脱水后冷冻，解冻后可使胚胎直接在ＰＢＳ中一步脱掉防冻剂，胚胎孵化率（４５．０％～４９．５％）与在０．２５ｍｏｌ／Ｌ蔗糖液中二步脱防冻剂的相似（５１．５％，Ｐ＞０．０５）。移植试验表明，在０．２５ｍｏｌ／Ｌ蔗糖液中二步脱防冻剂获得的妊娠率与六步脱防冻剂相似，与在ＰＢＳ中一步脱防冻剂的亦无明显差异，但妊娠率均偏低（２１．４％～３７．５％）。The effect of cryoprotectant being removed on the survival rate of frozen thawed day 7 8 bovine blastocysts produced in vitro was investigated. Dilution of cryoprotectants (0.75 mol/L glycerol + 0.5 mol/L propylene glycol) in 2 steps in 0.25 mol/L or 0.5 mol/L sucrose solution, or in 3 steps in 0.25 mol/L sucrose solution, resulted in a hatching rate similar to that of in 6 steps in PBS/FCS solution (68.6%, 62.2%, 68.7%, 70.4% respectively, P>0.05). Dilution in 3 steps in 0.5 mol/L sucrose solution significantly reduced the hatching rate (47.6%, P<0.01). Predehydration of embryos with 0.25 mol/L or 0.5 mol/L sucrose or trehalose solution allowed embryos to be diluted directly in PBS and gave a hatching rate (45.0% 49.5%) similar to that in 2 steps in 0.25 mol/L sucrose solution (51.5%, P>0.05). Embryo transfer trials showed that the pregnancy rate obtained from embryos diluted in 2 steps in 0.25 mol/L sucrose solution was similar to that obtained in 6 steps or 1 step dilution excepting the low pregnancy rate (21.4% 37.%).

关 键 词：体外受精 囊胚 防冻剂 孵化率 妊娠率 牛 

分 类 号：S823.36[农业科学—畜牧学]