人类成熟卵母细胞玻璃化冷冻研究

目的:研究人类成熟卵母细胞玻璃化的方法和冷冻液,探讨人类卵母细胞玻璃化冷冻保存的方法及临床应用价值。方法:运用cryoleaf进行人类成熟卵母细胞玻璃化冷冻保存,复苏后行胞浆内单精子注射-胚胎移植或培养至囊胚形成,观察复苏后卵母细胞受精及发育能力。根据冷冻方法分为程序化冷冻组和玻璃化冷冻组,又将玻璃化冷冻组分为自配液冷冻组和成品液冷冻组,比较各组存活率、受精率、卵裂率及囊胚形成率。结果:程序化冷冻组和玻璃化冷冻组间存活率、卵裂率、囊胚形成率均无显著性差异(P>0.05),但是受精率存在显著性差异(P<0.05);程序化组与自配液组、成品液组间存活率、卵裂率均无显著性差异(P>0.05),但受精率存在显著性差异(P<0.05)。成品液组与自配液组、程序化组间囊胚形成率存在显著性差异(P<0.05)。程序化组移植4例,获得临床妊娠并分娩1例,玻璃化组移植7例,获得1例生化妊娠。结论:程序化冷冻与玻璃化冷冻法均可应用于人类成熟卵母细胞,冻融后卵母细胞具备正常受精能力及卵裂能力。玻璃化冻存人类卵母细胞具有更大发展趋势。

Study on Vitrification of Human Mature Oocytes

Objective: To study the vitrification method and the cryopreservation solution of human mature oocytes, and to investigate the feasibility and clinical application value of oocytes cryopreservation. Methods: Human mature oocytes were cryopreserved using cryoleaf. Resuscitated oocytes were administered by intracytoplasmic sperm injection (ICSI), embryo transplantation or cultured to blastulation, observing the fertilization and developmental capacity of oocytes. The oocytes were divided into slow-freezing group and vitrification group, according to the cryopreservation methods. And then the vitrification group was divided into self-made up solution group and commercial solution group, and the survival rate, fertilization rate, cleavage rate and blastulation rate of all groups were compared. Result: The survival rate, cleavage rate and blastulation rate had no significant difference between slow-freezing group and vitrification group (P>0.05), while the fertilization rate had a significant difference (P<0.05). And there were significant difference of blastulation rate between commercial solution group and self-made up solution group (P<0.05). In slow-freezing group, embryos were transplanted to 4 patients, and one of them achieved clinical pregnancy and delivered a healthy boy. In vitrification group, embryos were transferred to 7 patients, and one of them got biochemical pregnancy. Conclusions: Both slow-freezing and vitrification methods can be applied to human mature oocytes, with the resuscitated oocytes having normal fertilizability and cleavage capability. And vitrification methods are more simple and convenient.