oocyte vitrification

卵母细胞玻璃化
  • 文章类型: Editorial
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  • 文章类型: Journal Article
    背景/目的:开放和封闭玻璃化系统通常用于卵母细胞冷冻保存;然而,关于比较它们对卵母细胞能力的单独影响的证据有限.这项研究独特地给文学带来了,关于开放式玻璃化与封闭式玻璃化系统对实验室和临床结果的影响的数据,以及冷却和升温速率的影响。方法:使用PubMed/MEDLINE数据库和Cochrane中央图书馆对文献进行了系统的检索,仅限于2023年1月之前以英文发表的文章。进行了网络荟萃分析,将每个玻璃化系统与新鲜卵母细胞进行了比较。结果:纳入23项研究。与新鲜卵母细胞相比,两种玻璃化装置均导致每个MII卵母细胞的受精率降低.当比较这两个系统的存活率时,差异无统计学意义。然而,有趣的是,与新鲜对照相比,开放系统导致每2个原核(2PN)卵母细胞的卵裂和胚泡形成率较低,同时,当将封闭设备与新鲜卵母细胞进行比较时,未检测到统计学上的显着差异。结论:总之,封闭的玻璃化系统似乎对卵母细胞的能力产生较少的有害影响,这反映在胚泡形成速率上。证明一个系统相对于另一个系统的优越性可能会导致标准化,有助于最终确定卵母细胞玻璃化的最佳实践。
    Background/Objectives: Open and closed vitrification systems are commonly employed in oocyte cryopreservation; however, there is limited evidence regarding a comparison of their separate impact on oocyte competence. This study uniquely brings to the literature, data on the effect of open versus closed vitrification systems on laboratory and clinical outcomes, and the effect of cooling and warming rates. Methods: A systematic search of the literature was performed using the databases PubMed/MEDLINE and the Cochrane Central Library, limited to articles published in English up to January 2023. A network meta-analysis was conducted comparing each vitrification system versus fresh oocytes. Results: Twenty-three studies were included. When compared to fresh oocytes, both vitrification devices resulted in lower fertilization rates per MII oocyte retrieved. When comparing the two systems in terms of survival rates, no statistically significant difference was observed. However, interestingly open systems resulted in lower cleavage and blastocyst formation rates per 2 pronuclear (2PN) oocyte compared to fresh controls, while at the same time no statistically significant difference was detected when comparing closed devices with fresh oocytes. Conclusions: In conclusion, closed vitrification systems appear to exert a less detrimental impact on the oocytes\' competence, which is reflected in the blastocyst formation rates. Proof of superiority of one system versus the other may lead to standardization, helping to ultimately determine optimal practice in oocyte vitrification.
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  • 文章类型: Journal Article
    目的:比较源自玻璃化卵母细胞的人类胚胎和源自新鲜收集卵母细胞的胚胎在卵母细胞捐献周期中的形态动力学模式。
    方法:一项回顾性观察性研究设置:胚胎移植生育诊所,胚胎学实验室,塞萨洛尼基,希腊患者(S):该研究包括58个卵母细胞捐赠周期的421个玻璃化卵母细胞和23个卵母细胞捐赠周期的196个新鲜卵母细胞的胚胎。
    方法:无主要结果度量(S):关键时间参数,动态事件,受精率,变性率,卵裂率,囊胚率,怀孕率,临床妊娠率,着床率和活产率。
    结果:玻璃化卵母细胞的存活率为92.58%(±7.42)。两组受精率差异显著(VITRI组:71.92%±20.29,对照组:80.65%±15.22,p=0.045),乳沟,胚泡,怀孕,临床妊娠,正在怀孕,来自新鲜或玻璃化卵母细胞的胚胎之间的着床率和活产率没有显着差异。延时分析显示,任何关键时间参数均无显着差异。然而,检查动态参数时,CC1[t2-tPB2:从第二极体挤出(tPB2)到2个细胞(t2)]显示出显着差异(p=0.004),而CC1a[t2-tPNf:从原核褪色(tPNf)到2个细胞(t2)]处于显着性阈值(p=0.057)。
    结论:与新鲜卵母细胞相比,玻璃化卵母细胞中的CC1表现出相对较慢的进展。相反,与新鲜卵母细胞相比,玻璃化卵母细胞中的CC1a表现出更快的进展。值得注意的是,这些临时偏差对后续发展影响很小。尽管玻璃化组的临床结果显示下降,均无统计学意义。这种意义的缺乏可以归因于有限的研究规模。
    OBJECTIVE: To compare the morphokinetic patterns of human embryos originating from vitrified oocytes (VITRI group) with those derived from freshly collected oocytes (CONTROL group) in oocyte donation cycles.
    METHODS: This is a retrospective observational study.
    METHODS: Embryolab Fertility Clinic, Embryology Lab, Thessaloniki, Greece.
    METHODS: The study included embryos from 421 vitrified oocytes from 58 oocyte donation cycles and 196 fresh oocytes from 23 oocyte donation cycles.
    METHODS: None.
    METHODS: Key time parameters, dynamic events, fertilization rates, degeneration rates, cleavage rates, blastocyst rates, pregnancy rates, clinical pregnancy rates, implantation rates, and live birth rates were estimated.
    RESULTS: The mean survival rate of vitrified oocytes was 92.58% (±7.42%). Fertilization rates were significantly different between the 2 groups (VITRI group: 71.92% ± 20.29% and CONTROL group: 80.65% ± 15.22%) whereas the degeneration, cleavage, blastocyst, pregnancy, clinical pregnancy, ongoing pregnancy, implantation, and live birth rates were not significantly different between embryos derived from fresh or vitrified oocytes. Time-lapse analysis showed no significant difference in any key time parameter. However, when examining dynamic parameters, first cell cycle (CC1) (t2 - tPB2: from the second polar body extrusion (tPB2) up to 2 cells (t2)) showed a significant difference whereas CC1a (t2 - tPNf: from fading of the pronuclei (tPNf) up to 2 cells (t2)) was at the threshold of significance.
    CONCLUSIONS: CC1 in vitrified oocytes exhibited a comparatively slower progression in contrast to fresh oocytes. Conversely, CC1a in vitrified oocytes demonstrated faster progression compared with fresh oocytes. It is worth noting that these temporary deviations had minimal impact on the subsequent development. Despite the clinical outcomes showing a decrease in the vitrified group, none of them reached statistical significance. This lack of significance could be attributed to the limited sample size of the study.
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  • 文章类型: Journal Article
    出于健康和社会原因,为不同年龄段的妇女提供卵母细胞冷冻保存。通过玻璃化冷冻保存来自受控卵巢刺激或体外成熟(IVM)的卵母细胞。由于母亲年龄是卵母细胞质量的重要决定因素,关于年龄相关的卵母细胞单独或联合使用玻璃化升温程序的易感性的数据有限。在本研究中,使用通过超排卵或IVM从2、6、9和12个月大的瑞士白化病小鼠获得的中期II卵母细胞。为了了解母亲年龄与卵母细胞冷冻耐受性之间的关系,对卵母细胞进行玻璃化加热,并与非玻璃化的同胞卵母细胞进行比较。评估存活的卵母细胞的线粒体电位,主轴完整性,纺锤体检查点蛋白转录本的相对表达,和DNA双链断裂。成熟潜能和玻璃化升温存活率在高龄组的排卵卵母细胞中受到显著影响(分别为p<0.001和p<0.05),而在IVM卵母细胞中没有影响。虽然玻璃化加温显著增加了高龄产妇排卵卵母细胞的纺锤体异常(p<0.01),在IVM卵母细胞中没有观察到显著的变化。此外,在玻璃化加温的IVM卵母细胞中,Bub1和Mad2转录物水平显著较高(p<0.05)。总之,高龄孕妇对小鼠排卵卵母细胞的冷冻敏感性有负面影响,但对小鼠IVM卵母细胞无影响。
    Oocyte cryopreservation is offered to women of various age groups for both health and social reasons. Oocytes derived from either controlled ovarian stimulation or in vitro maturation (IVM) are cryopreserved via vitrification. As maternal age is a significant determinant of oocyte quality, there is limited data on the age-related susceptibility of oocytes to the vitrification-warming procedure alone or in conjunction with IVM. In the present study, metaphase II oocytes obtained from 2, 6, 9, and 12 month old Swiss albino mice either by superovulation or IVM were used. To understand the association between maternal age and oocyte cryotolerance, oocytes were subjected to vitrification-warming and compared to non vitrified sibling oocytes. Survived oocytes were evaluated for mitochondrial potential, spindle integrity, relative expression of spindle checkpoint protein transcripts, and DNA double-strand breaks. Maturation potential and vitrification-warming survival were significantly affected (p < 0.001 and p < 0.05, respectively) in ovulated oocytes from the advanced age group but not in IVM oocytes. Although vitrification-warming significantly increased spindle abnormalities in ovulated oocytes from advanced maternal age (p < 0.01), no significant changes were observed in IVM oocytes. Furthermore, Bub1 and Mad2 transcript levels were significantly higher in vitrified-warmed IVM oocytes (p < 0.05). In conclusion, advanced maternal age can have a negative impact on the cryosusceptibility of ovulated oocytes but not IVM oocytes in mice.
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  • 文章类型: Journal Article
    不同条件下的生育力保护为妇女提供了购买时间的机会,这些时间可以投资于改善她们的福祉,通过从整体角度治疗他们的病情,符合现代医学的戒律。
    Fertility preservation for different conditions provides women the chance to buy time that can be invested in improving their well-being, by curing their condition from a holistic perspective, in line with the precepts of modern medicine.
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  • 文章类型: Journal Article
    不孕症是一种具有深刻社会影响的疾病。的确,医学和社会的发展都会影响IVF的实践方式,这并不奇怪。在现代医学的关键词是4P,含蓄地涉及(i)我们的知识和我们的技术的不断更新,以实现“预测”和“个性化”任务,(ii)鉴于所有相关的社会变革,不断重塑我们的思维方式,以完成“预防”和“参与”任务。全球老龄化人口的生活重点要求我们投资于生育教育,传播可操作的信息,让女性和男性做出有意义的生殖选择。在世界许多国家,出于医学和非医学原因的生育保护仍然非常被忽视,要求全面更新我们的方法,从学术界和试管婴儿实验室开始,穿过医疗办公室,接触(社交)媒体。生殖医学应该从治疗疾病的临床实践发展为保证患者生殖健康和福祉的整体方法。保存生育力的卵母细胞玻璃化是这种过渡的完美用例。这个工具正在获得一个新的身份,以符合新的适应症和社会需求,坚持技术挑战,全新的临床技术,和来自学术界的新革命。这篇“观点和评论”文章旨在从所有这些紧密联系的角度概述卵母细胞玻璃化的进展。
    Infertility is a condition with profound social implications. Indeed, it is not surprising that evolutions in both medicine and society affect the way in vitro fertilization is practiced. The keywords in modern medicine are the four principles, which implicitly involve a constant update of our knowledge and our technologies to fulfill the \"prediction\" and \"personalization\" tasks, and a continuous reshaping of our mindset in view of all relevant societal changes to fulfill the \"prevention\" and \"participation\" tasks. A worldwide aging population whose life priorities are changing requires that we invest in fertility education, spreading actionable information to allow women and men to make meaningful reproductive choices. Fertility preservation for both medical and nonmedical reasons is still very much overlooked in many countries worldwide, demanding a comprehensive update of our approach, starting from academia and in vitro fertilization laboratories, passing through medical offices, and reaching out to social media. Reproduction medicine should evolve from being a clinical practice to treat a condition to being a holistic approach to guarantee patients\' reproductive health and well-being. Oocyte vitrification for fertility preservation is the perfect use case for this transition. This tool is acquiring a new identity to comply with novel indications and social needs, persisting technical challenges, brand-new clinical technologies, and novel revolutions coming from academia. This \"views and reviews\" piece aims at outlining the advancement of oocyte vitrification from all these tightly connected perspectives.
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  • 文章类型: Journal Article
    卵母细胞在修复精子DNA损伤中起着至关重要的作用,这可能会影响下一代;然而,某些因素会削弱这种能力。这项研究检查了卵母细胞玻璃化,一种广泛使用的生育力保存方法,负面影响修复能力。雄性DBA/2小鼠(n=28)注射101.60μmol/100g体重的叔丁基过氧化氢(tBHP)14天以诱导精子DNA损伤。组织学变化,精子功能,使用TUNEL测定评估DNA片段化。使用Cryotop方法将超排卵的雌性DBA/2小鼠(n=28)的卵丘-卵母细胞复合物(COC)玻璃化。然后用tBHP处理和未处理的精子使新鲜和玻璃化的卵母细胞受精,和随后的胚胎发育被监测。此外,Mre11a的表达,使用实时PCR在所得受精卵和胚泡中评估Rad51、Brcal和Xrcc4。精子tBHP治疗减少了睾丸组织中分化的生精细胞,精子浓度,和运动性,同时增加DNA片段化(P<0.05)。经tBHP处理的精子玻璃化卵母细胞组的受精率降低(P<0.05),tBHP处理的精子新鲜和玻璃化卵母细胞组的双细胞率降低(P<0.05)。未处理的精子玻璃化卵母细胞和tBHP处理的精子新鲜和玻璃化卵母细胞组的四细胞囊胚率降低(P<0.05),tBHP处理的精子玻璃化卵母细胞组的囊胚率最低。在受精卵中,在tBHP处理的精子玻璃化卵母细胞组中Brca1上调(P<0.05)。此外,在胚泡中,在未处理的精子玻璃化卵母细胞组中,Rad51、Brca1和Xrcc4显著上调(P<0.05)。玻璃化对卵母细胞的损害会破坏精子DNA片段的修复,从而损害胚胎发育。
    Oocytes play a crucial role in repairing sperm DNA damage, which can affect the next generation; however, certain factors can impair this ability. This study examined whether oocyte vitrification, a widely used method for fertility preservation, negatively affects repair ability. Male DBA/2 mice (n = 28) were injected with 101.60 µmol/100 g body weight of tert-Butyl hydroperoxide (tBHP) for 14 days to induce sperm DNA damage. Histological changes, sperm functions, and DNA fragmentation were assessed using the TUNEL assay. Cumulus-oocyte-complexes (COCs) of superovulated female DBA/2 mice (n = 28) were vitrified using the Cryotop method. Fresh and vitrified oocytes were then fertilized by tBHP-treated and untreated sperms, and subsequent embryonic development was monitored. Additionally, the expression of Mre11a, Rad51, Brca1, and Xrcc4 was assessed in resulting zygotes and blastocysts using real-time PCR. The sperm tBHP treatment reduced differentiated spermatogenic cells in the testicular tissue, sperm concentration, and motility, while increasing DNA fragmentation (P < 0.05). The fertilization rate was decreased in the tBHP-treated sperm-vitrified oocyte group (P < 0.05), and the two-cell rate diminished in tBHP-treated sperm-fresh and vitrified oocyte groups (P < 0.05). The four-cell to blastocyst rate decreased in the untreated sperm-vitrified oocyte and the tBHP-treated sperm-fresh and vitrified oocyte groups (P < 0.05), and the tBHP-treated sperm-vitrified oocyte groups had the lowest blastocyst rate. In zygotes, Brca1 was upregulated in the tBHP-treated sperm-vitrified oocyte group (P < 0.05). Also, in blastocysts, Rad51, Brca1, and Xrcc4 were significantly upregulated in the untreated sperm-vitrified oocytes group (P < 0.05). Damages to the oocyte due to vitrification can disrupt the repair of sperm DNA fragmentation and consequently impair the embryo development.
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  • 文章类型: Journal Article
    玻璃化冷冻技术已广泛应用于卵母细胞冷冻保存,但仍需要优化以改善临床结局.在这项研究中,我们比较了常规液滴合并方案与改良的多梯度平衡玻璃化冷冻保存小鼠卵母细胞的中期II.随后,将卵母细胞解冻并进行卵胞浆内单精子注射(ICSI).通过形态学和免疫荧光染色评估卵母细胞的存活和纺锤体状态。此外,在体外检查受精率和囊胚发育。结果表明,多梯度平衡玻璃化在卵母细胞存活方面优于液滴合并玻璃化,纺锤形态,囊胚形成,和胚胎质量。相比之下,液滴合并玻璃化表现出降低的存活率,纺锤体形态正常的卵母细胞比例降低,随着加载卵母细胞数量的增加,胚泡率降低。值得注意的是,当超过六个卵母细胞被装载时,降低卵母细胞存活率,卵母细胞纺锤体形态异常,观察到胚胎质量差。这些发现突出表明,通过改良的多梯度平衡玻璃化对小鼠中期II卵母细胞的玻璃化具有维持卵母细胞存活的优势,纺锤形态,以及随后的胚胎发育。
    Vitrification has been widely used for oocyte cryopreservation, but there is still a need for optimization to improve clinical outcomes. In this study, we compared the routine droplet merge protocol with modified multi-gradient equilibration vitrification for cryopreservation of mouse oocytes at metaphase II. Subsequently, the oocytes were thawed and subjected to intracytoplasmic sperm injection (ICSI). Oocyte survival and spindle status were evaluated by morphology and immunofluorescence staining. Moreover, the fertilization rates and blastocyst development were examined in vitro. The results showed that multi-gradient equilibration vitrification outperformed droplet merge vitrification in terms of oocyte survival, spindle morphology, blastocyst formation, and embryo quality. In contrast, droplet merge vitrification exhibited decreasing survival rates, a reduced proportion of oocytes with normal spindle morphology, and lower blastocyst rates as the number of loaded oocytes increased. Notably, when more than six oocytes were loaded, reduced oocyte survival rates, abnormal oocyte spindle morphology, and poor embryo quality were observed. These findings highlight that the vitrification of mouse metaphase II oocytes by the modified multi-gradient equilibration vitrification has the advantage of maintaining oocyte survival, spindle morphology, and subsequent embryonic development.
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  • 文章类型: Journal Article
    卵巢子宫内膜异位症是一种与不孕症密切相关的妇科疾病-从其发病机理到治疗方式,这种情况在寻求受孕时对患者和临床医生都提出了挑战,由于AMH水平低,腹膜炎症,手术时无意中切除了健康的卵巢实质。事实上,大约一半的子宫内膜异位症患者寻求生育需要三级辅助生殖技术来实现活产.卵母细胞冷冻保存,最初为肿瘤患者设计的程序,近年来,对于被诊断患有卵巢子宫内膜异位症的患者来说,这是一种非常有希望的治疗策略,以保持其生育能力并在其生命的后期获得活产。在诊断的早期阶段咨询患者卵母细胞保存技术,最好是在35岁之前,尤其是在任何手术治疗之前,提供了一个很好的机会来讨论未来的生育能力和与卵母细胞冷冻保存相关的好处。
    Ovarian endometriosis is a gynecological condition that is closely associated with infertility-from its pathogenesis to treatment modalities, this condition presents a challenge both for patients and clinicians alike when seeking conception, due to low AMH levels, peritoneal inflammation, and the inadvertent removal of healthy ovarian parenchyma at surgery. In fact, around half of endometriosis patients seeking fertility require tertiary-level assisted reproduction techniques to achieve a live birth. Oocyte cryopreservation, a procedure initially designed for oncology patients, has emerged over recent years as a very promising treatment strategy for patients who have been diagnosed with ovarian endometriosis in order to preserve their fertility and obtain a live birth at a later stage in their lives. Counseling patients about oocyte preservation techniques at an early stage in the diagnosis, ideally before the age of 35 and especially prior to any surgical treatment, provides an excellent opportunity to discuss future fertility and the benefits associated with oocyte cryopreservation.
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  • 文章类型: Journal Article
    背景:特纳综合征(TS)伴有卵巢早衰。卵母细胞玻璃化是一种保存生育力的既定方法。然而,玻璃化卵母细胞的TS女性的卵母细胞产量数据以及利用卵母细胞的回报率很少。
    方法:回顾性多中心队列研究。数据是从2010年至2021年开始卵母细胞玻璃化的TS女性的医疗记录中收集的。
    结果:纳入了33名女性。玻璃化卵母细胞的累积数量中位数为每个妇女20个。并发症发生在4%的周期中。玻璃化卵母细胞的累积数量与AMH之间存在显着相关性(r=0.54,p<0.01)。AFC(r=0.49,p<0.01),46,XX细胞的百分比(r=0.49,p<0.01),和FSH(r=-0.65和p<0.01)。玻璃化后±3年,10名妇女发生了自发性(n=8)和IVF(n=2)妊娠。到目前为止,没有一个妇女返回使用他们的玻璃化卵母细胞。
    结论:对于患有TS的女性,卵母细胞玻璃化是一种可行的保留生育能力的选择,特别是那些有46,XX细胞系或足够卵巢储备的人。建议多次刺激周期以达到足够数量的玻璃化卵母细胞用于妊娠。现在得出关于TS女性使用玻璃化卵母细胞的结论还为时过早。
    BACKGROUND: Turner syndrome (TS) is accompanied with premature ovarian insufficiency. Oocyte vitrification is an established method to preserve fertility. However, data on the oocyte yield in women with TS who vitrify their oocytes and the return rate to utilize the oocytes are scarce.
    METHODS: Retrospective multicenter cohort study. Data was collected from medical records of women with TS who started oocyte vitrification between 2010 and 2021.
    RESULTS: Thirty-three women were included. The median cumulative number of vitrified oocytes was 20 per woman. Complications occurred in 4% of the cycles. Significant correlations were found between the cumulative number of vitrified oocytes and AMH (r = 0.54 and p < 0.01), AFC (r = 0.49 and p < 0.01), percentage of 46,XX cells (r = 0.49 and p < 0.01), and FSH (r = -0.65 and p < 0.01). Spontaneous (n = 8) and IVF (n = 2) pregnancies occurred in 10 women ± three years after vitrification. So far, none of the women have returned to utilize their vitrified oocytes.
    CONCLUSIONS: Oocyte vitrification is a feasible fertility preservation option for women with TS, particularly in those with 46,XX cell lines or sufficient ovarian reserve. Multiple stimulation cycles are recommended to reach an adequate number of vitrified oocytes for pregnancy. It is too early to draw conclusions about the utilization of vitrified oocytes in women with TS.
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