Non invasive assessment of human oocytes and embryos in assisted reproduction: Review on present practices and future trends

Oocyte and embryo grading is one of the most important steps in assisted reproductive technology to identify the best among cultured embryos for transfer or vitrification. The most commonly used non-invasive method is morphological assessment of gametes and embryos using a microscope. This method despite being simple and cost-effective has interobserver and intraobserver variability and subjectivity and has little to offer about the physiological state of embryos. This review sourced research articles and reviews pertaining to other non-invasive assessment methodologies from Medline and PubMed to collate latest technologies in vogue and identify novel methodologies of the future. The review assesses the current understanding in oocyte and embryo grading and touches upon novel non-invasive techniques and potential biomarkers to identify the best embryo. The latest available literature on time-lapse imaging, hierarchical algorithms, omics (consisting of proteomics and secretomes), miRNAs, mitochondrial RNAs and artificial intelligence has been accessed to summarize the enormous information available, to identify gaps in current interpretations, to identify emerging technologies and to provide direction for future research. This review will greatly benefit anyone practicing assisted reproduction and clinical embryology.     

Maternal benzo[a]pyrene exposure is correlated with the meiotic arrest and quality deterioration of offspring oocytes in mice

Benzo[a]pyrene (BaP) is a polycyclic aromatic hydrocarbon (PAH) in particulate matter that has a diameter of ≤2.5 μm (PM2.5). Studies have demonstrated that BaP exposure causes oocyte meiotic arrest in mice. However, whether BaP exposure also affects oocyte maturation in offspring remains unclear. To test this, female mice were administered BaP before pregnancy to generate BaP-exposed offspring. Our findings showed that BaP exposure reduced the in vitro maturation and increased the abnormalities of meiotic apparatus in offspring oocytes. In addition, BaP exposure reduced the mitochondrial content and intracellular ATP generation, induced early apoptosis, increased reactive oxidative species accumulation and the genomic DNA 5-methylcytosine (5mc) level in offspring oocytes. Along with the abovementioned defective parameters, maternal BaP exposure further compromised the embryo developmental competence of offspring oocytes. In summary, our study demonstrated that maternal BaP exposure compromised offspring oocyte maturation and quality.     

N-acetyl-l-cysteine protects porcine oocytes undergoing meiotic resumption from heat stress

Heat stress (HS) is a notable risk factor for female reproductive performance. In particular, impaired oocyte maturation was thought to contribute largely to the HS-induced reproductive dysfunctions. In this study, we confirmed that oocytes undergoing GVBD were much susceptible to HS, and thus compromising subsequent embryonic development. Using N-acetyl-l-cysteine (NAC), we found supplementation of a relatively high dose NAC during in vitro maturation, can protect oocytes from HS-induced complications, and thus rescuing impaired embryonic development. Further analysis indicated that mechanisms responsible for protecting GVBD oocytes from HS by NAC may include: (1) reversing disorganized spindle assembly and inhibited extracellular signal–regulated kinase (ERK) signaling; (2) correcting erroneous H3K27me3 modification and dysregulated expression of imprinted genes; (3) alleviating increased intraoocyte reactive oxygen species accumulation and apoptosis initiation. Our study, focusing on the oocyte meiotic maturation, may provide a safe and promising strategy for protecting reproductive sows under environmental hyperthermal conditions.     

Fertility preservation in patients of childbearing age treated for breast cancer: A nationwide cohort study

BackgroundApproximately 7% of breast cancers are diagnosed in women under 40. Question of subsequent fertility has become fundamental. We aimed to evaluate the rate of fertility preservation (FP) by oocyte retrieval (OR) after ovarian stimulation in patients of childbearing age, managed for breast cancer with adjuvant chemotherapy in France, reuse rate of frozen gametes and live births rate (LBR) after treatment.MethodsWe included 15,774 women between 18 and 40 years old, managed by surgery and adjuvant chemotherapy for breast cancer, between January 2011 and December 2020 from a French health registry. Patients with OR after breast surgery and before chemotherapy were considered as FP group; those with no OR as no FP group. To compare LBR with French population independently of age, we calculated Standardized Incidence Rates (SIR) of live births using indirect standardization method.ResultsFP rate increased gradually since 2011, reaching 17% in 2019. A decrease in use was observed in 2020 (13,9%). Among patients with at least 2 years of follow-up, gamete reuse rate was 5,6%. Births after cancer were mostly from spontaneous pregnancies. Among patients with at least 3 years of follow-up, LBR was 19,6% in FP group, 3,9% in second group. SIR of live births was of 1,05 (95% CI = 0.91–1.19) and 0.33 (95% CI = 0.30–0.36) in FP and no FP group respectively.ConclusionOncofertility activity increased until 2019 in France, reaching 17%. Gamete reuse rate was low. Births resulted mainly from spontaneous pregnancies. SIR of live births was lower in no FP group.     

Bovine cumulus-oocyte disconnection in vitro

Summary Cumulus-oocyte complexes were obtained from cows by aspiration of small (1–6 mm in diameter) antral follicles after slaughter. Complexes with a compact multilayered cumulus investment were cultured and processed for transmission electron microscopy after different periods of culture including a 0 h control group. In 0 h control oocytes the cumulus cells had numerous projections which penetrated the zona pellucida and established gap junctions with the oolemma. A partial loss of these junctions was noticed as an early event of oocyte maturation occurring within the first 3 h of culture. A low frequency of gap junctions was maintained until 12–18 h of culture where the junctional contact was completely disrupted. This decrease in intercellular communication was parallelled by resumption of oocyte meiosis.     

Fertility in patients with multiple sclerosis: current knowledge and future perspectives

Abstract The issue of fertility in patients with multiple sclerosis (MS) has not been exhaustively studied. Epidemiological data have suggested that spontaneous fecundity might be reduced; several endocrine and sexual disturbances potentially interfering with reproduction have been evidenced in MS patients of both sexes. Moreover, some medical treatments used in MS (e. g., mitoxantrone, cyclophosphamide) may exert detrimental effects on spermatozoa as well as on oocytes, leading to early impairment of fertility. This review illustrates the factors potentially interfering with fertility in MS and discusses the therapeutic tools that may be used to promote fertility in these patients. The safety of hormonal therapies in MS is also examined. The current applications of assisted reproductive technology (ART) are discussed, including in vitro fertilisation (IVF) techniques. Currently available methods to preserve fertility in patients that undergo cytotoxic treatments by means of sperm/oocyte cryostorage or by ovarian fragment cryopreservation and autografting are considered.     

降调节对卵子质量的影响

近10余年来，在体外受精一胚胎移植（IVF-ET）周期中应用促性腺激素释放激素激动剂（GnRH-a）进行控制性卵巢刺激（COS）已成为普遍采用的方法，其主要原因是GnRH-a可以促进卵巢内多个卵泡同步发育和成熟，抑制内源性黄体生成素（LH）峰，阻遏卵泡过早黄素化及卵子早熟。由于GnRH-a在IVF-ET进行降调节COS中的应用不同，可将COS方案分为长方案和短方案。[第一段]     

一氧化氮与卵巢生理

NO是一种结构简单的气体,是脊椎动物体内已发现的分子量最小的生物活性分子。NO作为一种信使分子,生物作用广泛,在生殖系统发挥重要作用。本文着重介绍NO作为卵巢内重要的调节因子,在调节卵泡发育、卵母细胞成熟、排卵、黄体的形成与退化、激素合成等方面的研究进展。     

Mechanism of action of the epileptogenic drug pentylenetetrazol on a cloned neuronal potassium channel

The action of the epileptogenic agent pentylenetetrazol (PTZ) on a cloned potassium channel of the rat brain was studied. The Kv1.1 channel was expressed in oocytes ofXenopus laevis and potassium currents were investigated in outside-out and inside-out membrane patches. The results show that PTZ increased the multi-channel potassium currents at strongly negative potentials and decreased them at potentials positive to −35 mV both in outside-out and inside-out membrane patches. The extent and manner of PTZ action, the concentration dependence as well as the onset and time course of the PTZ effect were the same both in outside-out and inside-out membrane patches. The single-channel potassium currents showed an increase in open probability and frequency of opening and a decrease in close time at −50 mV and vice versa at 0 mV with application of PTZ. The amplitude of single-channel current, the open time and the latency to the first channel opening remained almost unchanged under PTZ. The results indicate that PTZ acts via the cell membrane and influences the membrane-associated part of the potassium channel. Thereby, PTZ accelerates the transition from the inactivated to the open state of the channel at strongly negative potentials and reduces it at slightly negative and positive potentials. This mechanism may be the basis for a gate function which is in favour of the development of epileptic discharges.     

异丙酚麻醉不同阶段对卵细胞受精和对早期胚胎质量的影响

目的探讨异丙酚静脉麻醉下取卵手术中卵泡抽吸液中异丙酚的浓度变化及各阶段对卵细胞受精和早期胚胎质量的影响.方法 17例实施超促排卵下体外受精-胚胎移植术的妇女预测卵泡至少有15个,按取卵的先后分3组(早、中、晚)分别培养.记录从静脉给异丙酚到每个卵泡取出时间.检测卵泡抽取液中异丙酚的浓度及观察卵细胞的受精率及卵裂率等.结果取出第G一个卵泡的平均时间是200秒,取出卵泡的平均速率是17.6s/ 个.所有患者的最后一个卵泡抽吸液中异丙酚的浓度明显高于第一个卵泡抽吸液中异丙酚的浓度(P<0.01).除未成熟的卵细胞占总卵泡百分比在3组间有明显的区别外,受精率和卵裂率无明显区别.结论随着麻醉的进展,卵泡抽吸液中异丙酚的浓度逐渐增加,但与所给剂量或持续的时间之间无必然的关联.我们不能显示卵泡抽吸液中升高的异丙酚浓度对卵细胞质量的不利影响.