Recombinant anti-Müllerian hormone in the maturation medium improves the in vitro maturation of human immature (GV) oocytes after controlled ovarian hormonal stimulation.

Jure Bedenk, Tadeja Režen, Taja Železnik Ramuta, Nina Jančar, Eda Vrtačnik Bokal, Ksenija Geršak, Irma Virant Klun
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引用次数: 3

Abstract

Background: In vitro maturation (IVM) of oocytes is a laboratory method that allows the maturation of immature (GV) oocytes retrieved from patients enrolled in the in vitro fertilization (IVF) programme. However, this method is still sparsely researched and used in clinical practice, leading to suboptimal clinical results. Anti-Müllerian hormone (AMH) is an important hormone with known effects on human ovaries, especially on follicles (follicular cells) during folliculogenesis. In contrast, the effect of AMH on the human oocyte itself is unknown. Therefore, we wanted to determine whether human oocytes express AMH receptor 2 (AMHR2) for this hormone. Recombinant AMH was added to the IVM medium to determine whether it affected oocyte maturation.

Methods: In total, 247 human oocytes (171 immature and 76 mature) were collected from patients enrolled in the intracytoplasmic sperm injection (ICSI) programme who were aged 20 to 43 years and underwent a short antagonist protocol of ovarian stimulation. The expression of AMHR2 protein and AMHR2 gene was analysed in immature and mature oocytes. Additionally, maturation of GV oocytes was performed in vitro in different maturation media with or without added AMH to evaluate the effect of AMH on the oocyte maturation rate.

Results: Immunocytochemistry and confocal microscopy revealed that AMHR2 protein is expressed in both immature and mature human oocytes. AMHR2 was expressed in a spotted pattern throughout the whole oocyte. The IVM procedure revealed that AMH in maturation medium improved GV oocyte maturation in vitro, as all oocytes were successfully matured in maturation medium containing recombinant AMH only. Furthermore, antagonism between AMH and follicle-stimulating hormone (FSH) during the maturation process was observed, with fewer oocytes maturing when both AMH and FSH were added to the maturation medium. Finally, AMHR2 gene expression was found in immature and in vitro matured oocytes but absent in mature oocytes.

Conclusions: The positive AMHR2 protein and AMHR2 gene expression in human oocytes shows that AMH could directly act on human oocytes. This was further functionally confirmed by the IVM procedure. These findings suggest the potential clinical application of recombinant AMH to improve IVM of human oocytes in the future.

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在成熟培养基中加入重组抗勒氏杆菌激素,可促进受控卵巢激素刺激后人未成熟卵母细胞的体外成熟。
背景:卵母细胞体外成熟(IVM)是一种实验室方法,允许从参加体外受精(IVF)计划的患者中取出未成熟(GV)卵母细胞成熟。然而,该方法在临床实践中的研究和应用仍然很少,导致临床效果不理想。抗勒氏激素(AMH)是一种重要的激素,已知对人类卵巢,特别是在卵泡发生过程中对卵泡(滤泡细胞)有影响。相比之下,AMH对人类卵母细胞本身的影响尚不清楚。因此,我们想确定人类卵母细胞是否表达这种激素的AMH受体2 (AMHR2)。将重组AMH加入到IVM培养基中,观察其是否影响卵母细胞成熟。方法:共收集了247个人卵母细胞(171个未成熟卵母细胞和76个成熟卵母细胞),这些卵母细胞来自于年龄在20 - 43岁之间接受短期卵巢刺激拮抗剂方案的卵母细胞内单精子注射(ICSI)计划的患者。在未成熟和成熟卵母细胞中分析AMHR2蛋白和AMHR2基因的表达。此外,在体外不同成熟培养基中分别添加或不添加AMH进行GV卵母细胞成熟,以评估AMH对卵母细胞成熟率的影响。结果:免疫细胞化学和共聚焦显微镜显示,AMHR2蛋白在未成熟和成熟的人卵母细胞中均有表达。AMHR2在整个卵母细胞中呈斑点状表达。IVM程序显示成熟培养基中的AMH促进了GV卵母细胞的体外成熟,因为所有卵母细胞都在仅含有重组AMH的成熟培养基中成功成熟。此外,在成熟过程中观察到AMH和促卵泡激素(FSH)之间的拮抗作用,当成熟培养基中同时添加AMH和FSH时,成熟的卵母细胞较少。最后,AMHR2基因在未成熟卵母细胞和离体成熟卵母细胞中均有表达,而在成熟卵母细胞中无表达。结论:AMHR2蛋白和AMHR2基因在人卵母细胞中的阳性表达表明AMH可直接作用于人卵母细胞。IVM过程在功能上进一步证实了这一点。这些发现提示了未来重组AMH在改善人卵母细胞IVM方面的潜在临床应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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