Advanced Reproduction Medicine Research Center, Department of Obstetrics and Gynecology, International University of Health and Welfare School of Medicine.
Advanced Reproduction Medicine Research Center, Department of Obstetrics and Gynecology, International University of Health and Welfare School of Medicine;
J Vis Exp. 2021 Mar 25(169). doi: 10.3791/62098.
Ovarian function progressively declines during aging and in some pathophysiological conditions including karyotype abnormality, autoimmune diseases, chemo- and radiation-therapies, as well as ovarian surgeries. In unmarried women with severe ovarian dysfunction, fertility preservation is important for future pregnancies. Although oocyte cryopreservation is an established method for fertility preservation, these patients could only preserve a limited number of oocytes even after ovarian hyperstimulation, leading to repeated stimulations to ensure sufficient oocytes to guarantee future pregnancy. To solve this issue, we have recently developed a drug-free in vitro activation (IVA) procedure, which enable us to stimulate early stages of ovarian follicles to develop to the preantral follicle stage. These preantral follicles can respond to the unique protocol of gonadotropin stimulation, resulting in increased number of retrieved oocytes per ovarian stimulation for cryopreservation. The drug-free IVA comprised from the surgical approach and ovarian stimulation. We removed a part of cortex from one or both ovaries from patients under laparoscopic surgery. The ovarian cortical tissues were cut into small cubes to disrupt the Hippo signaling pathway and stimulate the development of early stage follicles. These cubes were grafted orthotropically into remaining ovaries as well as beneath the serosa of both Fallopian tubes. We have already published the surgical procedure of the drug-free IVA and the protocol of subsequent ovarian stimulation, but herein we present the details of laboratory methods required for drug-free IVA.
卵巢功能随着衰老以及某些病理生理状况而逐渐下降，这些状况包括核型异常、自身免疫性疾病、化疗和放疗，以及卵巢手术。对于严重卵巢功能障碍的未婚女性，生育力保存对于未来妊娠非常重要。虽然卵母细胞冷冻保存是生育力保存的一种既定方法，但这些患者即使在卵巢过度刺激后，也只能保存有限数量的卵母细胞，导致需要反复刺激以确保有足够的卵母细胞来保证未来妊娠。为了解决这个问题，我们最近开发了一种无药物的体外激活（IVA）程序，使我们能够刺激卵巢卵泡的早期阶段发育到原始卵泡阶段。这些原始卵泡可以对独特的促性腺激素刺激方案做出反应，从而在每次卵巢刺激中增加可用于冷冻保存的卵母细胞数量。无药物 IVA 包括手术方法和卵巢刺激。我们在腹腔镜手术下从患者的一侧或两侧卵巢中切除一部分皮质。将卵巢皮质组织切成小块，以破坏 Hippo 信号通路并刺激早期卵泡的发育。这些小块被原位移植到剩余的卵巢中，以及输卵管的浆膜下。我们已经发表了无药物 IVA 的手术程序和随后的卵巢刺激方案，但在此我们介绍了无药物 IVA 所需的实验室方法的详细信息。
