OP-IVM：将卵母细胞取回后的体外成熟与妇科手术相结合
1Center for Reproductive Medicine, Department of Obstetrics and Gynecology,Peking University Third Hospital, 2National Clinical Research Center for Obstetrics and Gynecology, 3Key Laboratory of Assisted Reproduction, Ministry of Education, 4Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproduction, 5Research Units of Comprehensive Diagnosis and Treatment of Oocyte Maturation Arrest,Chinese Academy of Medical Sciences
Summary
妇科手术前体外成熟（IVM）（OP-IVM）将卵母细胞取回后的IVM与常规妇科手术相结合，并作为常规IVM应用的延伸，用于保留生育能力。
Abstract
妇科手术前体外成熟（IVM）的使用（OP-IVM）是传统IVM的延伸，它将卵母细胞取回后的IVM与常规妇科手术相结合。OP-IVM适用于接受良性妇科手术的患者，他们需要保留生育能力（FP）或不孕症治疗，如体外受精和胚胎移植（IVF-ET）。在手术室中，接受良性妇科手术的患者首先被麻醉并接受超声引导的未成熟卵泡抽吸（IMFA）治疗。随着随后的妇科手术的进行，检查积云 – 卵母细胞复合物（COCs），并将未成熟的COCs转移到IVM培养基中并在IVF实验室中培养28-32小时。评估后，将选择MII阶段的成熟卵母细胞并将其冷冻保存在液氮中用于FP或通过卵胞浆内单精子注射（ICSI）受精用于IVF-ET。通过将IVM与妇科手术相结合，可以保存本来会丢弃的未成熟卵母细胞并将其用于辅助生殖技术（ART）。本文介绍了OP-IVM的程序，重要性和关键方面。
Introduction
IVM是一种ART，其中人未成熟的卵母细胞在体外培养至IVF-ET或FP成熟。在IVM中，不使用排卵诱导药物，从而减轻疼痛，经济负担和并发症，如卵巢过度刺激综合征（OHSS）1，2。此外，IVM特别适用于癌症患者的FP和不孕症患者的不孕症治疗，他们可能无法或没有时间接受排卵诱导治疗3。因此，虽然取回的卵母细胞数量、临床妊娠率（CPR）和活产率（LBR）均低于IVF4、5，但IVM有其独特的优势。
患有子宫内膜病变、输卵管积水或卵巢囊肿的不孕患者通常在 ART 治疗前进行妇科手术，其卵母细胞通常不成熟。OP-IVM使用引导经阴道超声来检索未成熟的卵母细胞并在体外生长，直到IVF-ET或FP成熟。OP-IVM在卵母细胞取回和妇科手术后结合IVM，从而减少受控卵巢过度刺激周期中常见的并发症，节省时间和金钱。对于有生育能力的患者，OP-IVM可以作为接受常规妇科手术的”生育保险”。
此外，妇科手术造成的损害，如电烙术6，7和卵巢肿瘤切除术，可以通过妇科手术前的卵母细胞取回来减少。因此，与常规妇科手术相比，OP-IVM可以减少不孕症治疗期间的手术次数，防止卵巢手术期间功能性卵母细胞的丧失。
先前的一项研究表明，额外的卵母细胞取回程序既不会增加手术并发症和不良妊娠结局，也不会延长住院时间8。一些患者通过OP-IVM8进行了活产，这表明这种方法的可行性。本文描述了可能受益于OP-IVM的患者的特征以及OP-IVM的程序和关键点，并讨论了人类卵母细胞成熟的评估。
Protocol
注：与OP-IVM方法相关的研究已获得北京大学第三医院机构审查委员会（IRB）和北京大学伦理委员会的批准（2014S2004）。OP-IVM的摘要如图 1所示。分步过程将在下一节中介绍。 1. 向合适的患者介绍OP-IVM 确定可能受益于OP-IVM的潜在患者，例如步骤1.1.1-1.1.3中描述的患者： 多囊卵巢综合征 （PCOS） 有克罗米芬耐药的患者，需要腹腔镜卵巢钻孔手?…
Representative Results
截至2019年12月，OP-IVM用于274名患者的生育能力保存。2014年至2016年间158名患者的胚胎学和生殖结局发表在之前的论文8中。以下示例讨论了2016年接受OP-IVM的PCOS患者所遵循的程序。患者为28岁，被诊断为原发性不孕、左附件囊肿和PCOS。她于2016年9月28日接受了腹腔镜膀胱切除术和OP-IVM手术。获得27个未成熟的COC。培养28小时后，选取7个MII阶段的卵母细…
Discussion
本文中描述的OP-IVM方法扩展到传统的IVM应用，并将卵母细胞取回后的IVM与常规妇科手术相结合。在妇科手术中会丢失的卵母细胞现在可用于IVF-ET或FP，而无需额外的手术风险。OP-IVM首先用于在PCOS患者卵巢钻孔之前取回卵母细胞。其应用很快扩展到需要良性妇科手术的不孕患者和需要放化疗的癌症或血液疾病患者。由于存在潜在的转移风险，OP-IVM不适合或推荐用于恶性肿瘤患者。基于OP-IVM，该机构?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
本研究由国家重点研发计划（编号：2017YFC1002000、2018YFC1004001、2019YFA0801400）、国家自然科学基金（编号：81571386、81730038）、CAMS医学创新基金（2019-I2M-5-001）和中国首都健康发展专项（2018-2-4095）资助。
Materials
| 19 G single-lumen aspiration needles | Cook, Australia | K-OPS-7035-REH-ET | |
| 4-well plate | Corning | ||
| 70 μm nylon cell strainer | Falcon, USA | 352350 | |
| CO2 Incubator | Thermo | ||
| Culture oil | Vitrolife, Sweden | 10029,OVOIL | Step 3.2. |
| FSH & LH | Ferring Reproductive Health, Germany | MENOPUR® | |
| Glass Pasteur pipette | Hilgenberg GmbH, Germany | 3154102-26 | |
| G-MOPS medium | pH-stable handling medium for washing the needle before puncturing | ||
| IVM medium | Origio, Denmark | ART-1600-B | |
| Laminar Flow Clean Benches | ESCO | ||
| Petri dish | Thermo Fisher Scientific, Denmark | 263991 | |
| pH stable handing media designed to support the handling and manipulation of oocytes and embryos outside the incubator | Vitrolife, Sweden | 10130, G-MOPS PLUS | Step 7.1. |
| Rinse solution | Cook, Australia | K-SIFB-100 | |
| Stereoscope | Nikon |
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Tags
OP-IVMIn Vitro MaturationOocyte RetrievalGynecological SurgeryInfertilitySubfertilityPolycystic Ovary SyndromeClomiphene ResistanceLaparoscopic Ovarian DrillingBenign Gynecologic SurgeriesHysteroscopic MyomectomyPolypectomyTransthoracic Resection Of SeptumLaparoscopic Tube SurgeryOophorocystectomyCancerHematologic DiseaseChemoradiotherapyRadiotherapyFertility PreservationUltrasound-guided AspirationFollicular Fluid AspirationHeparinCell StrainerGranulosa Cells
Cite This Article
Liu, T., Song, X., Zheng, X., Yan, L., Ma, C., Li, R., Yan, J., Qiao, J. OP-IVM: Combining In vitro Maturation after Oocyte Retrieval with Gynecological Surgery. J. Vis. Exp. (171), e61647, doi:10.3791/61647 (2021).