牛体外受精胚胎一步细管冷冻解冻技术的研究

本试验探讨 1.4M、1.0M或 0 .7M三个不同浓度的甘油冷冻牛体外受精胚胎 ,解冻采用 0 .5M或0 .2 5M蔗糖液细管内一步除甘油的方法。结果 6种组合中 ,以 1.0M甘油配 0 .5M蔗糖为最好 ,同 1.4M甘油配 0 .2 5M或 0 .5M蔗糖和 0 .7M甘油配 0 .5M蔗糖这三种组合比 ,胚胎的孵化率差异显著 (84 .35 %vs6 6 .6 7%、71.4 3%、73.98% ,P <0 .0 5 ) ;与另两个组合比 ,胚胎的孵化率差异不显著 (84 .35 %vs 75 .0 %、75 .4 % ,P >0 .0 5 )。三个甘油浓度之间 ,1.0M的冷冻效果最好 ,而 1.4M的效果最差 ,且二者之间胚胎的孵化率差异显著 (79.6 5 %VS 6 9.4 8% ,P <0 .0 5 )。以上结果表明 ,不同甘油浓度对冷冻牛体外受精胚胎有明显的影响。细管内的组合以含 1.0M甘油为冷冻液 ,两端装 0 .5M蔗糖为最佳

Freezing IVF Bovine Embryos by One-step Dilution of Cryoprotectants

Dilution of cryoprotectants for freezing of IVF bovine embryos was investigated in this study. Day 7 blastocysts were frozen in 1.4M, 1.0M or 0.7M glycerol (G) using the conventional freezing procedure. After thawing of embryos, glycerol was removed by one-step dilution in 0.25M or 0.5M sucrose (S). When they were frozen in 1.0MG and diluted in 0.5MS significantly more embryos hatched (84.35%) in comparison with them frozen in 1.4MG and diluted in 0.5M (66.67%) or o.25MS(71.43%), and frozen in 0.7MG and diluted in 0.5MS(73.98%). The hatching rate of embryos frozen in 1.0MG (79.65%) was significantly higher (p<0.05) than that frozen in 1.4MG(69.48%). This result indicates that the concentration of glycerol significantly influence the survival rate of bovine IVF embryos after frozen-thawed. Appropriate concentration of glycerol for embryo freezing and sucrose for glycerol removed are 1.0M and 0.5M, respectively.