人黄体期卵母细胞的体外成熟及玻璃化冷冻的实验研究

Research on the In Vitro Maturation and Vitrification of Oocytes from Human Luteal Phase

目的:探讨未成熟卵母细胞体外成熟(IVM)技术联合玻璃化冷冻保存黄体期卵母细胞对某些女性肿瘤患者的生育能力的保存情况。方法:采集因妇科肿瘤等行卵巢切除手术过程中穿刺获取的256枚未成熟卵母细胞,按取卵时患者的月经周期分为卵泡期组(143枚)与黄体期组(113枚),每组再随即分为新鲜对照组及玻璃化冷冻组。分别进行IVM后行新鲜卵胞质内单精子注射(ICSI)授精和玻璃化冻融卵ICSI授精,比较各组间IVM后MII卵率、受精率、卵裂率、优质胚胎率。结果:①卵泡期与黄体期卵母细胞的IVM率差异无统计学意义;而组间复苏存活率(68.0%vs 48.1%)有统计学差异(P<0.05);卵泡期与黄体期卵母细胞新鲜组间受精率、卵裂率、优质胚胎率无统计学差异,冷冻组间差异亦无统计学意义。②与新鲜组相比,玻璃化冷冻使卵泡期与黄体期卵母细胞的受精率均降低(P<0.01)。结论:黄体期未成熟卵母细胞可以体外成熟并有继续发育为优质胚胎的能力;玻璃化冷冻使卵母细胞受精率、卵裂率下降。IVM和冻融后体外授精是某些女性肿瘤患者保存生育能力的一种有临床应用前景的方式。

Objective： To investigate in vitro maturation（IVM）,vitrification-thawing and ICSI of immature oocytes from human luteal phase as to preserve the fertility of some female cancer patients.Methods： A total of 256 immature oocytes were aspirated from 64 women undergoing gynecologic operations.They were divided by menstrual cycle into follicular pahase group and luteal phase group,morethan each group was divided into virtrification subgroup and fresh subgroup（the control）.Maturation（MII stage） rate,fertilization rate,cleavage rate and rate of good-quality embryos were observed.Results： 1） There was no significant difference in MII stage rate between oocytes retrieved at luteal phase and follicular phase（P〈0.05）.The frozen-thawen survival rate for the oocytes retrieved was higher at follicular phase（68.0%,34/63） than at luteal phase（48.1%,26/38）（P〈0.05）.The fertilization rate,cleavage rate and rate of good-quality embryos showed no significant difference between the immature oocytesof folliular phase and luteal phase.2） Compared with fresh group,cryopreservation showed significantly decline the fertilization rate in both of the oocytes from follicular phase and luteal phase（P〈0.01）.Conclusion： Immature oocytes retrieved at luteal phase can be matured in vitro and develop to good-quality embryos;vitrification declines the fertilization rate and cleavage rate of oocytes.In vitro maturation,vitrification-thawing and intracytoplasmic sperm injection is a convenient way for female cancer patients to preserve their fertility in the clinical work.