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提高体外采集的人类卵母细胞在卵巢组织冷冻保存过程中的成熟率。

Improving the maturation rate of human oocytes collected ex vivo during the cryopreservation of ovarian tissue.

机构信息

Laboratory of Reproductive Biology, The Juliane Marie Centre for Women, Children and Reproduction, Copenhagen University Hospital and Faculty of Health and Medical Sciences, University of Copenhagen, 2100, Copenhagen, Denmark.

Università degli studi di Teramo, 64100, Teramo, Italy.

出版信息

J Assist Reprod Genet. 2020 Apr;37(4):891-904. doi: 10.1007/s10815-020-01724-7. Epub 2020 Feb 24.

Abstract

PURPOSE

The aim of the present study was to improve the in vitro maturation (IVM) procedure using oocytes from surplus ovarian tissue after fertility preservation.

METHODS

Twenty-five patients aged 17-37 years were included in the study. Maturation was compared between oocytes collected in HEPES-buffered medium or saline, and we determined whether transport on ice prior to oocyte collection affected maturation. Two different IVM media were used that were supplemented with and without recombinant human midkine. Mature oocytes were assessed for aneuploidy using next-generation sequencing (NGS).

RESULTS

On average, 36 immature oocytes were collected from each patient (range 7-90, N = 895). Oocytes recovered from HEPES-buffered medium matured at a higher rate than oocytes recovered from saline (36% vs 26%, p < 0.01). Ovarian transportation on ice prior to the procedure negatively affected maturation compared with non-transported samples (42% vs 27%, p < 0.01). The addition of midkine improved maturation rate (34% vs 27%, p < 0.05). On average, 11 MII oocytes were obtained per patient (range 1-30). NGS of 53 MII oocytes and their first polar bodies indicated that 64% were euploid.

CONCLUSIONS

The study demonstrated unexpectedly high number of immature oocytes collected from surplus ovarian tissue without any stimulation. The overall MII rate was one in three, resulting in a total number of MII oocytes that was similar to the number obtained after ovarian stimulation. If these MII oocytes prove suitable for IVF, they will provide a substantial improvement in fertility preservation for patients and advance IVM as an interesting platform for further improvements in assisted reproduction.

摘要

目的

本研究旨在通过对接受生育力保存的剩余卵巢组织中的卵母细胞进行体外成熟(IVM)来提高该过程。

方法

本研究纳入了 25 名 17-37 岁的患者。比较了在 HEPES 缓冲液或盐水中采集的卵母细胞的成熟情况,并确定在采集卵母细胞之前是否通过冰上运输影响了成熟。使用两种不同的 IVM 培养基,分别补充和不补充重组人中期因子。使用下一代测序(NGS)评估成熟卵母细胞的非整倍体性。

结果

每位患者平均从卵巢组织中采集 36 个未成熟卵母细胞(范围 7-90,N=895)。与从盐水中回收的卵母细胞相比,从 HEPES 缓冲液中回收的卵母细胞成熟率更高(36%比 26%,p<0.01)。与未运输的样本相比,在手术前将卵巢冰上运输会对成熟产生负面影响(42%比 27%,p<0.01)。添加中期因子可提高成熟率(34%比 27%,p<0.05)。每位患者平均获得 11 个 MII 卵母细胞(范围 1-30)。对 53 个 MII 卵母细胞及其第一极体进行 NGS 分析表明,64%的卵母细胞为整倍体。

结论

该研究出乎意料地从剩余卵巢组织中采集到大量未成熟卵母细胞,且无需任何刺激。总的 MII 率为三分之一,这导致获得的 MII 卵母细胞总数与卵巢刺激后获得的卵母细胞数量相似。如果这些 MII 卵母细胞适合 IVF,将为患者的生育力保存带来实质性改善,并将 IVM 作为进一步提高辅助生殖的有趣平台。

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