添加两种不同培养成分对人未成熟卵体外成熟培养的影响

目的:研究添加两种不同培养成分对人未成熟卵母细胞体外成熟培养(IVM)的效果。方法:选择接受ICSI治疗的77名不孕患者共收集未成熟卵177枚,随机分A组(激素组)33例,添加0.075 IU/ml FSH及0.075 IU/ml LH至基础IVM培养液中;B组(成熟卵泡液组)44例,50%基础IVM培养液与50%人成熟卵泡液配成。将两组收集的未成熟卵母细胞放入不同培养液中培养48 h,每24 h倒置显微镜下观察卵母细胞的形态;对MII卵进行ICSI,ICSI操作后继续培养,ICSI后16～20 h进行受精观察,24及48 h分别进行卵裂观察及胚胎评分。结果:①B组未成熟卵母细胞48 h成熟率和MI期卵母细胞48 h成熟率显著高于A组,差异有统计学意义(P<0.05),两组培养液GV期卵母细胞24和48 h成熟率比较差异无统计学意义(P>0.05)。②B组2PN卵裂率显著高于A组,差异有统计学意义(P<0.05),两组2PN受精率及可利用胚胎率差异无统计学意义(P>0.05)。结论:人成熟卵泡液内可能含有一些未知的有利于卵母细胞成熟的因子,深入研究卵泡液的成分,进一步明确其促卵母细胞成熟机制对开发新的IVM培养液、改善IVM的临床结局有重要意义。

Effects of two kinds of different adding ingredients on in vitro maturation of human immature oocytes

Objective:To research the effects of two kinds of different adding ingredients on in vitro maturation(IVM) of human immature oocytes.Methods:A total of 77 infertile patients who received intracytoplasmic sperm injection(ICSI) were selected,and then 177 immature oocytes were collected;77 patients were randomly divided into two groups: in group A(33 patients,hormone group),0.075 IU/ml follicle-stimulating hormone(FSH) and 0.075 IU/ml luteinizing hormone(LH) were added into basic IVM culture solution;in group B(44 patients,mature follicular fluid group),the culture solution was composed of 50% basic IVM culture solution and 50% human mature follicular fluid.The immature oocytes in the two groups were cultured in different culture solutions for 48 hours,the morphologies of oocytes were observed under inverted microscope every 24 hours;MII oocytes were treated with ICSI,then they were cultured continuously,fertilization was observed at 16-20 hours after ICSI,cleavage was observed and embryo score was performed at 24 and 48 hours after ICSI.Results:The 48-hour mature rates of immature oocytes and MII oocytes in group B were statistically significantly higher than those in group A(P<0.05),there was no statistically significant difference in 24-hour and 48-hour mature rates of oocytes at GV stage between the two groups(P>0.05).The cleavage rate of 2PN in group B was statistically significantly higher than that in group A(P<0.05),there was no statistically significant difference in 2PN fertilization rate and rate of available embryos between the two groups(P>0.05).Conclusion:Human mature follicular fluid may contain some cytokines beneficial to maturation of oocytes,further researching the component of follicular fluid and confirm the mechanism of oocytes maturation have important significances for developing new IVM culture solutions and improve clinical outcome of IVM.