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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。
Objective: To study the effects of adding two different culture components on in vitro maturation of human immature oocytes (IVM). Methods: A total of 177 immature oocytes were collected from 77 infertile patients receiving ICSI. A total of 177 immature eggs were collected and randomly divided into group A (hormone group), 33 cases, 0.075 IU / ml FSH and 0.075 IU / ml LH into basal IVM culture medium. B group (mature follicular fluid group) 44 cases, 50% basic IVM broth and 50% human mature follicular fluid dubbed. The immature oocytes collected in the two groups were cultured in different culture medium for 48 h, and the morphological changes of oocytes were observed under inverted microscope every 24 h. ICSI and ICSI of MII eggs were continued after culture, and after ICSI 16-20 h fertilization observation, cleavage observation and embryo score 24 and 48 h respectively. Results: (1) The 48 h maturation rate of immature oocytes and the 48 h maturation rate of MI oocytes in group B were significantly higher than those in group A (P <0.05). The GV oocytes 24 There was no significant difference in 48 h maturation rate (P> 0.05). ② The 2PN cleavage rate in group B was significantly higher than that in group A (P <0.05). There was no significant difference in fertilization rate and available embryo rate between the two groups (P> 0.05). CONCLUSION: The human mature follicular fluid may contain unknown factors that are conducive to oocyte maturation, and further study the composition of follicular fluid to further clarify the mechanism of oocyte maturation for the development of new IVM broth, to improve the clinical outcome of IVM There’s important meaning.