In Vitro Oocyte Maturation Techniques

体外卵母细胞成熟技术
  • 文章类型: Review
    McCune-Albright综合征(MAS)是一种罕见的遗传性疾病,影响多个器官,包括内分泌组织。这种内分泌病有时会导致不孕症,因为它可能诱导卵巢的独立功能,导致无排卵周期。该病例报告描述了一名22岁女性的不孕症旅程,该女性青春期早,雌激素和孕激素水平高,不规律,低FSH和LH(在月经周期的第3天),和多囊性右卵巢。她接受了几种不孕症治疗:最初是体外卵母细胞成熟(IVM),然后是囊肿经阴道超声引导的抽吸,都不成功。进行了右半卵巢切除术,最终恢复了规律的周期,并有可能进行卵巢刺激(OS)和体外受精(IVF)。在第一次胚胎移植后获得活产。
    McCune-Albright syndrome (MAS) is a rare genetic disease affecting multiple organs, including endocrine tissues. This endocrinopathy is sometimes responsible for infertility, as it may induce an independent functioning of the ovaries leading to anovulatory cycles. This case report describes the infertility journey of a 22-year-old female who had early puberty and irregular periods with high estrogen and progesterone levels, low FSH and LH (on day 3 of her menstrual cycle), and a multi-cystic right ovary. She received several infertility treatments: initially in vitro oocyte maturation (IVM) followed by cyst transvaginal ultrasound-guided aspiration, all unsuccessful. A right hemi-ovariectomy was performed that eventually restored regular cycles and made it possible to perform ovarian stimulation (OS) and in vitro fertilization (IVF). Live birth was obtained after the first embryo transfer.
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  • 文章类型: Case Reports
    背景:抵抗性卵巢综合征(ROS)是一种罕见的内分泌疾病,很少有关于患病患者活产的报道。由于促性腺激素抗性导致未成熟卵母细胞,一些研究人员报道了很少有卵母细胞体外成熟(IVM)的活产,但在ROS患者中,IVM并不总是成功的。这里,我们报告了一个ROS的原始病例,与血清中的Ig-FSHR相关,卵巢刺激联合地塞米松治疗后实现活产。
    方法:30岁女性出现继发性闭经和不孕症。发现她的血清FSH水平高于正常水平,但与正常的抗苗勒管激素(AMH)水平和窦卵泡计数不一致。遗传调查未发现可能影响FSHR的突变。参考以前的ROS研究,分析患者血清中针对FSHR的抗体,斑点印迹分析显示与FSHR的反应性强.然后,向患者推荐了地塞米松,她成功地怀孕了,最终通过剖腹产接生了一个健康的女孩.
    结论:据我们所知,这是首次报道卵巢刺激联合地塞米松成功治疗ROS.在某些情况下,ROS大剂量外源性促性腺激素联合免疫抑制治疗可能是一种有效的方法。
    BACKGROUND: Resistant ovary syndrome (ROS) is a rare endocrine disorder and there have been few reports of live births by affected patients. As gonadotropin resistance leads immature oocytes, some researchers reported few live births with in vitro maturation (IVM) of oocytes, but IVM is not always successful in ROS patients. Here, we report an original case of ROS, associated with Ig-FSHR in the serum, who achieved a live birth following ovarian stimulation combined with dexamethasone treatment.
    METHODS: The 30-year-old woman presented with secondary amenorrhea and infertility. Her serum FSH levels were found to be higher than normal, but in discordance with a normal anti-Müllerian hormone (AMH) level and antral follicle count. Genetic investigation found no mutations potentially affecting FSHR. With reference of previous ROS studies, the patient\'s serum was analyzed for antibodies directed against FSHR and dot blot analysis showed strong reactivity with FSHR. Then, dexamethasone was proposed to the patient, and she successfully became pregnant, finally delivering a healthy girl by caesarean section.
    CONCLUSIONS: To our best knowledge, this is the first report of the successful treatment of ROS using ovarian stimulation combined with dexamethasone. In some cases of ROS, high doses of exogenous gonadotropins in combination with immunosuppressive therapy could be an effective approach.
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  • 文章类型: Case Reports
    目的:我们报道了一名自身免疫性卵巢早衰(POF)患者在卵母细胞体外成熟(IVM)和新鲜胚胎移植后成功的活产,并对该适应症中卵母细胞IVM治疗后获得的活产文献进行了综述。
    方法:患者是一名24岁的女性,产后诊断为自身免疫性POF,患有血清抗卵巢和抗21-羟化酶抗体的自身免疫性多腺综合征。患者有典型的POF症状:继发性闭经伴雌激素减少,促性腺激素升高和不育;然而,血清抗苗勒管激素水平和总窦卵泡计数保持正常。在连续三天给予150IU高度纯化的人绝经促性腺激素并注射10,000IU人绒毛膜促性腺激素以触发排卵后,进行未成熟卵母细胞的IVM。
    结果:收集的6个卵母细胞-卵丘复合体在体外成熟。卵胞浆内单精子注射(ICSI)后,获得5个胚胎。在两个胚胎的新鲜移植和适当的子宫内膜准备后实现妊娠。妊娠37周后分娩了一名正常的女性儿童,其特征是肾上腺功能不全和不稳定的糖尿病。
    结论:我们报告了一例自身免疫性卵巢早衰(POF)患者在IVM治疗后成功的活产。具有自身免疫性病理学的育龄妇女的管理需要生育咨询。自身免疫性POF的早期诊断对于早期受孕和保存卵母细胞具有重要意义。因为目前唯一的选择是卵子捐赠。
    OBJECTIVE: We report a successful live birth after oocytes in vitro maturation (IVM) and fresh embryo transfer in a patient with autoimmune premature ovarian failure (POF) and performed a review of the literature of livebirths obtained after oocytes IVM treatment in this indication.
    METHODS: The patient was a 24-year-old woman with autoimmune POF diagnosed post-partum, who developed autoimmune polyglandular syndrome with serum anti-ovarian and anti-21-hydroxylase antibodies. The patient had typical symptoms of POF: secondary amenorrhea with hypoestrogenism, elevated gonadotropins and infertility; however, the serum anti-Müllerian hormone level and total antral follicle count remained normal. IVM of immature oocytes was performed after the administration of 150 IU highly purified human menopausal gonadotropin for three consecutive days and an injection of 10,000 IU human chorionic gonadotropin to trigger ovulation.
    RESULTS: The six oocyte-cumulus complexes collected matured in vitro. After intracytoplasmic sperm injection (ICSI), five embryos were obtained. Pregnancy was achieved after the fresh transfer of two embryos and appropriate endometrial preparation. A normal female child was delivered following a 37-weeks pregnancy characterized by the onset of adrenal insufficiency and unstable diabetes.
    CONCLUSIONS: We report a successful livebirth after IVM treatment in a patient with autoimmune premature ovarian failure (POF). Management of reproductive age women with autoimmune pathology requires fertility counseling. Early diagnosis of autoimmune POF is important for early conception and oocyte preservation, because the only other option at present is ovum donation.
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  • 文章类型: Case Reports
    OBJECTIVE: Assisted oocyte activation combined with ICSI (ICSI-AOA) has been reported to improve fertilization outcomes of couples with oocyte activation deficiency (OAD). Although there\'s no sufficient evidence to support ICSI-AOA as routine use, it might be beneficial for POSEIDON group 3 patients with suspected oocyte-related OAD.
    METHODS: A 29-year-old female presented with a history of primary infertility for two years. She was classified as a POSEIDON group 3 patient and had a total fertilization failure history. With the help of ICSI-AOA, six oocytes were successfully fertilized. Pregnancy was later confirmed after embryo transfer. A living infant was born after 34 weeks of pregnancy.
    CONCLUSIONS: OAD should be taken into consideration for POSEIDON group 3 patients since low Antimüllerian hormone is associated decreased quality. Further research needs to be done to understand the mechanism underlying oocyte-related OAD and the potential role of ICSI-AOA in young patients with suboptimal ovarian response.
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  • 文章类型: Case Reports
    目的:本研究旨在报道一例在没有卵巢刺激的情况下收集未成熟卵母细胞的肿瘤患者紧急保留生育力的病例,通过体外成熟(IVM),随后是ICSI和受精卵的冷冻保存,十年后,在一个健康婴儿的活产中。
    方法:2008年9月,我们的诊所对一名32岁的女性进行了IVM,该女性被诊断为具有阳性雌二醇受体的导管浸润性癌,孕激素受体阴性和人表皮生长因子受体2阳性。在简单的乳房切除术后和任何化学疗法治疗之前,在没有卵巢刺激的情况下进行未成熟卵母细胞的回收。将收集的致密卵丘-卵母细胞复合物(COCs)置于Lag培养基中2小时,然后在IVM培养基中孵育,补充热灭活的患者血清,重组FSH,和重组LH。培养30小时后,去除卵丘细胞,中期II卵母细胞被显微注射,将获得的受精卵冷冻保存。2017年,将受精卵解冻并培养至第3天。一个胚胎被转移,另一个被冷冻保存。
    结果:收集四个紧凑型COC并进行IVM。两个卵母细胞到达中期II并进行显微注射。获得了两个受精卵,并在两个原核阶段冷冻保存。大约9年后,将两个受精卵解冻并培养至第3天。移植并植入了10个细胞的胚胎,导致一个健康的婴儿出生。
    结论:在紧急开始辅助治疗需要立即收集卵母细胞的情况下,IVM可能是获得完全合格的成熟卵母细胞的唯一选择,从而可以有效保存生殖潜力。
    OBJECTIVE: This study aims to report a case of urgent fertility preservation in an oncological patient with collection of immature oocytes in the absence of ovarian stimulation that, through in vitro maturation (IVM), followed by ICSI and cryopreservation of zygotes resulted, 10 years later, in the live birth of a healthy baby.
    METHODS: In September 2008, our clinic performed IVM in a 32-year-old woman diagnosed with a ductal invasive carcinoma with positive estradiol receptors, negative progesterone receptors and positive human epidermal growth factor receptor 2. The retrieval of immature oocytes was performed in the absence of ovarian stimulation after a simple mastectomy and prior to any chemotherapy treatment. The compact cumulus-oocyte complexes (COCs) collected were placed in Lag medium for 2 h, followed by incubation in IVM medium, supplemented with heat inactivated patient serum, recombinant FSH, and recombinant LH. After 30 h in culture, cumulus cells were removed, the metaphase II oocytes were microinjected, and the zygotes obtained were cryopreserved. In 2017, the zygotes were thawed and cultured until day 3. One embryo was transferred and the other cryopreserved.
    RESULTS: Four compact COCs were collected and subjected to IVM. Two oocytes reached metaphase II and were microinjected. Two zygotes were obtained and were cryopreserved at the two pronuclear stage. Approximately 9 years later, the two zygotes were thawed and cultured until day 3. An embryo with 10 cells was transferred and implanted, resulting in the birth of a healthy baby.
    CONCLUSIONS: In cases where urgency to start adjuvant therapy requires immediate oocyte collection, IVM may be the only option to obtain fully competent mature oocytes allowing for effective preservation of the reproductive potential.
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  • 文章类型: Journal Article
    随着性腺毒性肿瘤治疗后稳定缓解率的增加,为了为肿瘤患者提供最佳护理,需要新的生育能力保存方法。这里,我们报道了1例卵巢组织卵母细胞体外成熟(OTOIVM)后的整倍体囊胚冷冻保存的原始病例。乳腺癌患者卵巢切除术后,从卵巢组织中获得了33个卵母细胞。12个成熟卵母细胞中有6个成功受精,形成3个囊胚。对与这种恶性肿瘤相关的突变的遗传调查发现,患者不是携带者。仅对非整倍体进行了植入前遗传测试,发现所有3个胚泡都是整倍体,适合胚胎移植。我们的研究表明,体外成熟的卵巢组织卵母细胞在ICSI后具有形成整倍体囊胚的潜力,可以筛选非整倍体和遗传突变,然后玻璃化,以便为癌症女性提供最佳的生育力保护策略。
    With the increased rate of stable remission after gonadotoxic cancer treatment, new methods of fertility preservation are required in order to provide the best possible care for oncological patients. Here, we report an original case of euploid blastocyst cryopreservation after in vitro maturation of ovarian tissue oocytes (OTO IVM). Thirty-three oocytes were obtained from the ovarian tissue after ovariectomy in the breast cancer patient. Six out of 12 matured oocytes fertilized successfully and 3 blastocysts were formed. Genetic investigation for mutations associated with this type of malignancy found that the patient is not a carrier. Preimplantation genetic testing was performed only for aneuploidies and found all 3 blastocysts to be euploid and suitable for embryo transfer. Our study showed that the ovarian tissue oocytes matured in vitro have the potential for euploid blastocyst formation after ICSI which could be screened for aneuploidies and inherited mutations and then be vitrified in order to provide the best fertility preservation strategy for women with cancer.
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  • 文章类型: Case Reports
    BACKGROUND: Although women with polycystic ovarian syndrome (PCOS)-related sub-fertility are high responders to controlled ovarian stimulation, it is difficult to obtain mature oocytes in these women. Therefore, in vitro maturation (IVM), which is the technique of letting the contents of the ovarian follicles and the oocytes inside mature in vitro, has often been proposed in such women. We describe the first successful delivery of monozygotic triplets resulting from transfer of a single blastocyst following IVM of oocytes.
    METHODS: A 32-year-old nulligravida female with PCOS underwent IVM. She underwent vitrified-warmed single blastocyst transfer following IVM, and a dichorionic triamniotic triplet pregnancy was confirmed at 8 weeks. Healthy triplets were delivered by cesarean section at 33 weeks\' gestation. This is the first case of monozygotic triplets derived from IVM oocytes that were successfully delivered. The determination of chorionicity and amnionicity is generally supposed until 3 days after fertilization, and no division or splitting of her embryo was observed on transfer. Interestingly, her embryo might have split after the transfer, resulting in a dichorionic triamniotic triplet pregnancy.
    CONCLUSIONS: Patients should be informed of a possible increased risk of monozygotic multiple pregnancies after single embryo transfer following IVM.
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  • 文章类型: Case Reports
    Here, we present a diffuse large B cell lymphoma patient admitted for fertility preservation before cancer therapy and whose pregnancy was recognized incidentally just after the start of random start controlled ovarian stimulation (RSCOH) during the stimulation cycle. Despite an optimal homogenous growth of follicle cohort, majority of the retrieved oocytes were immature after GnRHa trigger. Possible effects of extremely high serum progesterone and/or β-hCG levels on oocyte in vivo maturation are discussed with the surprising high rate of in vitro maturation and subsequent good embryo development. It seems that in case of need for pregnancy termination as a result of an urgent cancer therapy, RSCOH can be started and patients may benefit from overnight in vitro maturation of oocytes.
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  • 文章类型: Case Reports
    Infertility due to Gonadotropin-Resistant Ovary Syndrome (GROS) is a rare type of hypergonadotropic hypogonadism. Here, we report an original case of GROS, associated with compound heterozygous follicle-stimulating hormone receptor (FSHR) variants, in a woman who achieved a live birth by in vitro maturation (IVM) of her oocytes. This 31-year-old woman consulted our assisted reproduction center for a second opinion after having been advised, because of pervasive high serum follicle-stimulating hormone (FSH) levels, to pursue in vitro fertilization (IVF) with donor oocytes. She presented with primary infertility and progressively prolonged menstrual cycles. Her serum FSH levels were indeed found to be high, but in discordance with a normal anti-Müllerian hormone (AMH) level and antral follicle count. Genetic investigation found the patient to be compound heterozygous for two FSHR variants: I160T, a known pathologic variant, and N558H, which has never been previously reported. As there was no ovarian response to high daily doses of exogenous gonadotropins, IVM was proposed to the patient with success and she finally delivered at term a healthy boy. Effects of the receptor variants were analyzed in heterologous cells. Whereas the I160T mutation blocked FSHR membrane trafficking and FSH-stimulated cAMP-dependent signaling in transfected CHO cells, the novel variant, N558H, functioned equivalently to wild-type FSHR in the assays employed. In conclusion, IVM should always be offered as a first-line therapy to infertile women presenting with GROS. The N558H variant discovered in FSHR is novel, but its functional significance, if any, is unresolved and merits further investigation as it may be associated with a recessive FSHR-related disorder.
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  • 文章类型: Case Reports
    The purpose of ovarian stimulation in IVF is to recover mature oocytes at metaphase II stage which are capable of fertilization either when mixed with sperm or after ICSI. However, there have been instances when even after controlled ovarian stimulation (COS) and correct administration of human chorionic gonadotrophin (hCG) trigger for final oocyte maturation, the oocytes were found to be arrested at germinal vesicle (GV) or metaphase I (MI) stage. Similar dilemma is faced in cases of empty follicle syndrome (either genuine or due to inadequate response), however, in this condition, there is no retrieval of oocytes despite presence of mature looking follicles. We present an interesting case where despite presence of normally growing follicles and documentation of correct response to trigger and rise in estradiol levels, two subsequent IVF cycles; one triggered with recombinant hCG and second with GnRH agonist, hCG failed to yield mature oocytes. Both cycles yielded expected number of oocytes but all at immature MI stage even after dual trigger.
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