不同载体和冷冻保护剂用于人胚胎玻璃化冷冻的初步研究

目的：比较分析冷冻环、自制叶片、冰珠3种载体，以及5．5mol／LEG、15％PROH＋15％EG2种冷冻保护剂用于人胚胎玻璃化冷冻的效果。方法：①选择131个6-10细胞，Ⅰ-Ⅱ级胚胎，按载体不同随机分为3组，将胚胎解冻后用于移植，比较3组的复苏率、植入率、临床妊娠率；②部分患者成功妊娠分娩后，自愿放弃剩余冻存胚胎，选择6-10细胞，Ⅰ-Ⅱ级，以叶片为载体冷冻的胚胎52个，按照冷冻溶液的不同分为2组，解冻后将胚胎继续培养3天，比较2组的复苏率和囊胚形成率。结果：①冰珠组的复苏率显著低于冷冻环和自制叶片组，3个组的植入率、临床妊娠率无明显差异；②15％PROH＋15％EG冻存的胚胎解冻后复苏率和囊胚形成率显著高于5．5mol/L EG。结论：冷冻环和自制叶片是较为理想的冷冻载体，二者用于玻璃化冷冻效果相近，自制叶片具有成本低的优点；联用15％PROH＋15％EG能降低冷冻保护剂对胚胎的毒性，玻璃化冷冻效果好于5．5mol／LEG。

A Pilot Study on Different Carriers and Cryoprotectants Applied for Human Embryos Vitrification

Objective:To compare the effects of three different carriers(cryoloop,self-made blade and ice-ball)and two different cryoprotectants(5.5mol/L EG and 15%PROH+15%EG)on human embryos vitrification.Methods:①131 embryos (6-10 cells,Ⅰ-Ⅱgrade)were randomly divided into 3 groups according to different carriers.The embryos were used for transplantation after thawing,and the recovery rate,implantation rate,and clinical pregnancy rate were compared.②Some patients with successful pregnancy and childbearing gave up the remaining vitrified embryos voluntarily.And 52 embryos (6-10 cells,Ⅰ-Ⅱgrade)carried on blade were divided into 2 groups according to different cryoprotectants.The embryos were cultured for 3 days after thawing.The recovery rate and blastulation rate were compared.Results:①The recovery rate of ice-ball group was significantly lower than cryoloop and self-made blade groups.The differences of implantation rate and clinical pregnancy rate were not statistically significant among the 3 groups.②The recovery rate and blastulation rate of 15% PROH+15%EG group were significantly higher than 5.5mol/L EG group.Conclusion:Cryoloop and self-made blade are relatively ideal vitrification carriers with similar effects.Self-made blade has the advantage of lower cost.The combination of 15%PROH+15%EG can achieve a better effect than 5.5mol/L EG for its lower cryoprotectant toxicity on embryos.