Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China.
Fertil Steril. 2011 Sep;96(3):629-634.e1. doi: 10.1016/j.fertnstert.2011.06.054. Epub 2011 Jul 16.
To investigate the effect(s) of exogenous gonadotropin on the cytoplasmic and nuclear maturation of cumulus-partially enclosed immature human oocytes in vitro derived from ovarian stimulation cycles.
Experimental human study.
University-based laboratory.
PATIENT(S): Women, aged 26-35 years, with infertility secondary to male factors, underwent ovarian stimulation and intracytoplasmic sperm injection cycles using a long protocol of pituitary down-regulation.
INTERVENTION(S): Cumulus-partially enclosed immature human oocytes that were retrieved from the stimulated cycles were collected at the time of intracytoplasmic sperm injection. The cumulus-partially enclosed immature human oocytes were allocated into two groups: [1] oocytes at the germinal vesicle (GV) stage; and [2] oocytes at the metaphase I (MI) stage. Each group was cultured in vitro with and without gonadotropin supplements. Some metaphase II (MII) oocytes derived from the two groups were parthenogenetically activated and exposed to subsequent embryonic development for 168 hours in vitro. Other MII oocytes were tested for meiotic apparatus analysis, including spindle morphology and chromosomal alignment, by immunofluorescence staining and scanning confocal microscopy.
MAIN OUTCOME MEASURE(S): Oocyte maturation and activation rates, percentages of embryonic development, and spindle normalization were analyzed by χ(2) analysis, whereas oocyte maturation time was analyzed by one-way analysis of variance.
RESULT(S): For GV oocytes the maturation and activation rates were significantly higher during in vitro maturation with supplementation with FSH/LH (68% vs. 60% and 82% vs. 62%, respectively). However, maturation time (22.78 ± 0.87 vs. 23.70 ± 0.94 hours), embryonic development (cleavage: 84% vs. 83%; four-cell: 72% vs. 66%; eight-cell: 48% vs. 43%; blastocyst: 5% vs. 7%), and meiotic apparatus normalization rates (55% vs. 61.1%) were similar. For MI oocytes there were no significant differences in the maturation rates (85% vs. 84%), maturation time (14.81 ± 0.65 vs.15.73 ± 0.58 hours), activation rates (77% vs. 80%), embryonic development (cleavage rates: 80% vs. 83%; four-cell: 68% vs. 72%; eight-cell: 56% vs. 51%; blastocyst: 7% vs. 6%), and meiotic apparatus normalization rates (52.4% vs. 54.5%). CONLUSION(S): Gonadotropin supplements to the maturation medium play an important role in cumulus-partially enclosed oocytes at the GV stage; however, MI stage-derived oocytes from stimulated cycles fail to acquire improved maturity after in vitro maturation. Furthermore, gonadotropin at the current concentration did not increase spindle or chromosomal abnormalities in MII oocytes maturated from either GV- or MI-stage oocytes in vitro.
研究外源性促性腺激素对体外来源于卵巢刺激周期的卵丘部分包裹的不成熟人卵母细胞的细胞质和核成熟的影响。
实验性人体研究。
大学实验室。
年龄 26-35 岁，因男性因素导致不孕，接受了长垂体下调方案的卵巢刺激和胞浆内精子注射周期。
在胞浆内精子注射时收集来自刺激周期的卵丘部分包裹的不成熟人卵母细胞。将卵丘部分包裹的不成熟人卵母细胞分为两组：[1]处于生发泡（GV）阶段的卵母细胞；[2]处于第一次减数分裂中期（MI）阶段的卵母细胞。每组均在有和没有促性腺激素补充的情况下进行体外培养。从这两组中获得的一些第二次减数分裂中期（MII）卵母细胞通过孤雌激活，并在体外进一步进行 168 小时的后续胚胎发育。其他 MII 卵母细胞通过免疫荧光染色和扫描共聚焦显微镜检查分析减数分裂装置，包括纺锤体形态和染色体排列。
卵母细胞成熟和激活率、胚胎发育率以及纺锤体正常化率通过卡方检验进行分析，而卵母细胞成熟时间通过单因素方差分析进行分析。
GV 期卵母细胞在体外成熟时添加 FSH/LH 后，其成熟和激活率显著提高（分别为 68% vs. 60%和 82% vs. 62%）。然而，成熟时间（22.78±0.87 小时 vs. 23.70±0.94 小时）、胚胎发育（卵裂：84% vs. 83%；四细胞：72% vs. 66%；八细胞：48% vs. 43%；囊胚：5% vs. 7%）和减数分裂装置正常化率（55% vs. 61.1%）相似。对于 MI 期卵母细胞，成熟率（85% vs. 84%）、成熟时间（14.81±0.65 小时 vs. 15.73±0.58 小时）、激活率（77% vs. 80%）、胚胎发育率（卵裂率：80% vs. 83%；四细胞率：68% vs. 72%；八细胞率：56% vs. 51%；囊胚率：7% vs. 6%）和减数分裂装置正常化率（52.4% vs. 54.5%）均无显著差异。
促性腺激素补充剂对 GV 期卵丘部分包裹的卵母细胞成熟具有重要作用；然而，来自刺激周期的 MI 期卵母细胞在体外成熟后未能获得改善的成熟度。此外，目前浓度的促性腺激素并未增加体外成熟的 GV 期或 MI 期卵母细胞获得的 MII 卵母细胞中的纺锤体或染色体异常。
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