Cryoloop和CPS超速玻璃化冷冻人三原核胚胎效果比较

Comparison of Two Ultra-rapid Vitrification Techniques for Human Three-pronuclear Embryos: Cryoloop and Closed Pulled Straw Group(CPS).

目的:比较冷冻环(Cryoloop)和闭合型拉细麦管(CPS)超速玻璃化冷冻方法冻存人早期卵裂期胚胎的效果差异,以期建立1种有效的超速玻璃化冷冻方法。方法:将人三原核孕卵发育形成的126个6-10细胞胚胎随机分为3组:对照组30个、Cryoloop组36个和CPS组60个。通过形态学观察,比较胚胎复苏后存活率;经罗丹明123染色,激光扫描共聚焦显微镜摄片分析,测定复苏后存活胚胎线粒体跨膜电位(△Ψm)。结果:Cryoloop组复苏后胚胎存活率达100%,显著高于CPS组(P<0.05);Cryoloop组△Ψm显著高于CPS组(P<0.05),而与未冷冻对照组相比,无明显差异(P>0.05)。结论:Cryoloop超速玻璃化冷冻法获得了更高的胚胎存活率和胚胎活力,该方法更适于冷冻人早期卵裂期胚胎。

Objective： To compare the results of cryoloop and closed pulled straw vitrification methods for the purpose of establishing an effective ultra - rapid vitrification method for human cleavage embryos. Methods： The 6 - 10 cell human embryos derived from three - pronuclear embryos were divided into three groups randomly, 30 in control group , 36 in cryoloop group and 60 in CPS group. At first, the survival rates were evaluated through the embryo morphology after thawing. Then the survival embryos were stained with Rhodamine123 to examine the mitochondrial membrane potential under the laser confoeal microscope （LSCM） . Results： Compared to CPS group, signific, ant higher embryo survival rates were obtained in the cryoloop group. There was no difference in embryo survival rates between cryoloop group and control group. The mitochondrial membrane potential in CPS group is lower than cryoloop group, the difference between eryoloop group and control group was uaremarkable. Compared to the control group, the mitochondrial membrane potentials in CPS group was significant lower. Conclusion： The cryoloop ultra -rapid vitrification technique not only obtained higher embryo survival rate but also conserved the embryos vitality. It is more effective to cryopreserve early human cleavage embryos.