玻璃化冷冻活检后人胚胎的探索

The First Attempt at Vitrification of Biopsied Human Cleavage Stage Embryos

目的:探索玻璃化方法冷冻活检后人胚胎的可行性。方法:实验组中活检后的胚胎先后在含10%乙二醇平衡液和30%乙二醇加0.5mol/L蔗糖的玻璃化液中进行平衡,然后将胚胎置于冷冻薄膜上,直接在液氮中快速冷冻。复温时采用1mol/L浓度的蔗糖液。对照组中活检后的胚胎进行标准程序化丙二醇慢冻。结果:采用玻璃化冷冻技术后,活检后的人胚胎冻存率、完整率、卵裂球总体存活率、囊胚形成率均显著比使用标准程序化丙二醇慢冻方法提高(95%对15%,P<0.01;75%对9%,P<0.01;89%对24%,P<0.01;20%对2%,P<0.05)。结论:玻璃化冷冻可以显著提高活检后的卵裂期的人胚胎的冻存结局。

Objective： To find an effective method to improve cryosurvival of biopsied human embryos. Methods： In study group, biopsied embryos were pre-equilibrated by 10% EG（ethylene glycol）solution, vitrified in 30% ＋ 0.5 mol/L solution, loaded on the cryotops and frozen directly in liquid nitrogen, lmol/L sucrose was adopted in the warming media. In control group, the biopsied embryos were cryopreserved by standard programmed slow propanediol protocol. Results： The cryosurvival rate, intact rate, total blastomere survival rate, blastulation rate were all significantly higher using the vitrification method than using the standard slow propanediol method （95% vs 15%, P〈0.01; 75% vs9%, P〈0.01; 89% vs24%, P〈0.01; 20% vs2%, P〈0.05）. Conclusions. Outcomes of cryopreservation of human biopsied cleavage stage embryos can be improved significantly by vitrification.