Study On The Cryopreservation Of Mice And Human Oocytes By The Vitrification Method

Objective: Study on the cryoprotective effect of the EDS-40 vitrification solutions to mice and human oocytes. Methods: 1.We have devised EDS-40 vitrification solution ourself.According to random group doctine,the test is divided into experiment group and contrast group . Experiment group(a) use self-devised EDS-40 vitrification solution(including 40% Ethylene glycol and 18% Dextran) , contrast group(b) use EFS-40 vitrification solution which is considered the best vitrification solution. and contrast group(c) use program freezing method.First to test the vitrification effect of the EDS-40 vitrification solution and EFS-40 vitrification solution .2. Animal experiment:,In group a and group b , good kunnming mice oocytes were vitrified in EFS-40 and EDS-40 solutions at 25℃,after packaged in 0.25ml plastic straws ,the oocytes were immersed into liquid nitrogen ,in group c good kunnming mice oocytes were cryopresevered in program freezing equipment. All samples were cryopreserved about 2-3 months. The straws were warmed rapidly in 25℃ water, and agitated gently, then oocytes were transferred into Ham'sF12 medium and cultured. Then watch its survival rate and its fertilization rates. Software SPSS11.5 analysis was used to determine differences in percentage effect. 3.Human experiment: Human oocytes were collected from our center,the cryopreserved method ,rewarming method and statistical analysis method is same to animal experiment . Result: 1.EDS-40 solutions were vitrified.2. The toxicity test of two vitrification solutions had significant difference.3.the survival rate,of mice oocytes were79.34%,67.94%,56.34%;the fertilization rate of mice oocytes were79.17%,66.29%,51.14%。4.The survival rate of human oocytes were53.57 %,66.67%,56.67 % and the fertilization rate of human oocytes were40.00%,77.78%,41.18%. Conclusion: 1. EDS-40 vitrification can be verified and can be the cryoprotectant to vitrifiy oocytes . 2.The effect of EDS-40 solution is beteer than EFS-40 and program freezing method on cryopreserving mice oocytes.3. Our experiment outcome manifest the effect of vitrification is better than conventional protocol. 4. The effect of EDS-40 solution is lest than EFS-40 and similar to program freezing method on cryopreserving humanoocytes,besides the species difference ,the cause is related to the quality of embryos,and need tcontinue to study.