自然周期体外受精取卵前卵母细胞提早排出的预测因素分析

目的分析卵巢低反应患者自然周期体外受精(IVF)方案中发生取卵前卵母细胞提早排出的相关因素,探讨预测和预防卵母细胞在取卵前提早排出的相关指标。方法实施自然周期IVF/卵胞浆内单精子注射(ICSI)治疗的患者共378个周期,自月经第3天B超监测窦卵泡直径,基础卵泡刺激素(FSH)、黄体生成索(LH)、雌二醇(E_2)值;第8¹1天B超监测卵泡直径;当卵泡直径>14 mm时,监测血LH、E_2、孕酮(P)水平,当E_2≈1,100 pmol/L时,肌注0.2 mg促性腺激素释放激素激动剂(GnRH-a)诱发排卵(trigger),3411天B超监测卵泡直径;当卵泡直径>14 mm时,监测血LH、E_2、孕酮(P)水平,当E_2≈1,100 pmol/L时,肌注0.2 mg促性腺激素释放激素激动剂(GnRH-a)诱发排卵(trigger),34³6 h后取卵。分析各项临床指标与卵母细胞在取卵前提早排出发生的相关性。结果取卵前卵母细胞提早排出与基础E_2值、"trigger"日P、LH值有关,与患者年龄、基础LH、FSH值和卵泡直径无关。结论自然周期取卵时机的选择应根据患者基础E_2值及监测P、LH水平进行预测和监控。     

Melatonin accelerates maturation inducing hormone (MIH): induced oocyte maturation in carps

The present communication is an attempt to demonstrate the influence of melatonin on the action of maturation inducing hormone (MIH) on the maturation of oocytes in carps. The oocytes from gravid female major carp Labeo rohita were isolated and incubated separately in Medium 199 containing (a) only MIH (1 microg/ml), (b) only melatonin (at concentrations of 50, 100 or 500 pg/ml), and (c) both melatonin and MIH, but at different time intervals. In the latter group, melatonin was added to the incubating medium either (i) 4 h before addition of MIH, (ii) 2 h before addition of MIH, (iii) co-administered with MIH (0 h interval) or (iv) 2 h after addition of MIH. In each case, oocytes were further incubated for 4, 8, 12 or 16 h post- administration of MIH, and the effects of treatment on oocyte maturation were evaluated by considering the rate (%) of germinal vesicle breakdown (GVBD). Incubation of oocytes in a medium containing only melatonin did not result in GVBD of any oocyte. Nearly all the oocytes underwent GVBD when incubated with MIH for 16 h. Administration of melatonin along with MIH (at 0 h interval) or 2 h after addition of MIH did not result in any significant change in the rate of GVBD compared to that in a medium containing only MIH. However, it was quite interesting to observe that incubation of oocytes with melatonin especially 4 h prior to addition of MIH in the medium, led to an accelerated rate of GVBD in the oocytes. Experiments with the oocytes of another major carp Cyprinus carpio following an identical schedule depicted similar results except a difference in the optimum melatonin dose. In L. rohita, 50 pg/ml melatonin had maximum acceleratory effect on MIH-induced GVBD of oocytes, while it was 100 pg/ml in C. carpio. Further study revealed that pre-incubation with melatonin accelerates the action of MIH on the formation of a complex of two proteins (MPF), a regulatory component called cyclin B and the catalytic component protein kinase known as cyclin-dependent kinase, Cdk1. Densitometric analysis of the immunoblot data collected from the melatonin pre-treated MIH incubated oocytes showed that cyclin B level continued to increase even after 4 h of incubation, and reached the peak after 12 h. Moreover, determination of H1 kinase activity as an indicator of MPF activity in oocytes revealed that melatonin pre-incubation considerably increased MIH stimulation of histone H1 phosphorylation as compared to MIH alone. Thus, the present study demonstrates for the first time that prior incubation with melatonin accelerates the action of MIH on carp oocyte maturation.     

人卵母细胞体外成熟的影响因素探讨

目的　提高人未成熟卵母细胞体外培养的成熟率。方法　将机械法获取的人未成熟卵母细胞随机分为观察组与对照组进行体外培养 ,观察影响卵母细胞体外成熟的主要影响因素。结果　年龄 <30岁者未成熟卵母细胞的体外成熟率明显高于 >30岁者 (P<0 .0 5 ) ;观察组上清液中表皮生长因子 (EGF)、血管内皮生长因子(VEGF)和粒细胞 -巨噬细胞集落刺激因子 (GM- CSF)的含量明显高于对照组 (P<0 .0 5 )。结论　 EGF、VEGF和GM- CSF可提高卵母细胞体外培养的成熟率 ;年龄也是影响卵母细胞体外成熟的重要因素     

Cloning of zebrafish ovarian carbonyl reductase-like 20 beta-hydroxysteroid dehydrogenase and characterization of its spatial and temporal expression

20 beta-Hydroxysteroid dehydrogenase (20 beta-HSD) is a crucial enzyme that converts 17 alpha-hydroxyprogesterone to 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (DHP), which triggers oocyte maturation in most teleost fish. A full-length cDNA for a carbonyl reductase-like 20 beta-HSD (CR/20 beta-HSD) has been cloned from the zebrafish ovary. Although the zebrafish CR/20 beta-HSD is expressed in all of the tissues tested, it is predominantly expressed in the ovary, testis, kidney, and gill. In the ovary, the enzyme was shown to be expressed in the follicle cells and its expression appeared to be constitutive. No significant difference was noticed in the level of CR/20 beta-HSD expression among follicles of different stages. Furthermore, analysis of the ovarian samples taken at different times before spawning showed no significant change of the enzyme expression. In agreement with these results, treatment of the cultured zebrafish ovarian follicle cells with gonadotropin and activin had little effect on the expression of the enzyme. Taken together, these results point to the possibility that the gonadotropin-induced DHP production and final oocyte maturation in the zebrafish may not involve significant change of CR/20 beta-HSD expression as evidenced in the salmonids, or that there might be other isoforms of 20 beta-HSD whose expression is tightly controlled by endocrine and paracrine factors.     

Role of gap junctions and protein kinase A during the development of oocyte maturational competence in Ayu (Plecoglossus altivelis)

Meiotic resumption in teleost oocytes is induced by a maturation-inducing hormone (MIH). The sensitivity of oocytes to MIH, also known as oocyte maturational competence (OMC), is induced by LH via mechanisms that are not fully understood. A previous study of Ayu (Plecoglossus altivelis) showed the presence of functional heterologous gap junctions (GJs) between oocytes and their surrounding granulosa cells. The objectives of this study were to determine the role of ovarian GJs and of protein kinase A (PKA) during the acquisition of OMC. We examined the effects of the specific GJ inhibitor carbenoxolone (CBX) and 18alpha-glycyrrhetinic acid (alpha-GA) on the LH-(hCG)-dependent acquisition of OMC and on MIH-(17,20beta-dihydroxy-4-pregnen-3-one)-dependent meiotic resumption; measured the cAMP content of ovarian follicles during the hCG-dependent acquisition of OMC; and determined the effects of PK activators and inhibitors on hCG-dependent OMC. Production of follicular cAMP increased during the hCG-dependent acquisition of OMC. Both GJ inhibitors and the PKA inhibitor H8-dihydrochloride, but not the PKC inhibitor GF109203X, suppressed the hCG-dependent acquisition of OMC in a dose-dependent manner. The PKA activator forskolin induced OMC with a similar potency to hCG. Unlike previous observations with teleosts where disruption of heterologous GJ either blocks or stimulates meiotic resumption, treatment with GJ inhibitors did not affect MIH-dependent meiotic resumption in maturationally competent follicles of Ayu. These observations suggest that ovarian GJs are essential for LH-dependent acquisition of OMC but not for MIH-dependent meiotic resumption, and that the stimulation of OMC by LH is mediated by cAMP-dependent PKA. They are also consistent with the view that a precise balance between GJ-mediated signals (positive or negative) and oocyte maturational readiness is required for hormonally regulated meiotic resumption.     

Optimum oocyte retrieved and transfer strategy in young women with normal ovarian reserve undergoing a long treatment protocol: a retrospective cohort study

Purpose

This study aimed to investigate the relationship between the number of oocytes retrieved and clinical outcomes in young women with normal ovarian reserve who were undergoing their first in vitro fertilization and embryo transfer (IVF-ET) cycle. The transfer strategy based on yielded oocytes was also discussed in this article.

Methods

A total of 1567 patients who underwent first long protocol of IVF treatment in our reproductive medical center between January 2010 and June 2014 were categorized into five groups based on the retrieved oocyte number, namely, 4∼6, 7∼9, 10∼12, 13∼15, and ≥16. Baseline parameters were similar among the groups. Primary outcome was defined as the cumulative live birth rate (CLBR), and secondary outcomes included the rate of patients with high risks for ovarian hyperstimulation syndrome (OHSS).

Results

It was found that the CLBR increased with the number of oocytes, as well as the rate for high risks of OHSS. In fresh cycles, 10∼12 oocyte group demonstrated the highest implantation rate (53.32 %), clinical pregnancy rate (CPR) (73.13 %), and live birth rate (LBR) (61.14 %), with no significant differences. Moreover, both cumulative CPR (CCPR) and CLBR became significantly higher in the 10∼12 oocyte group, compared with 4∼6 and 7∼9 groups. However, when the retrieved oocytes increased to 13∼15 or ≥16, the cumulative results did not have a significant increase. Also, the high risk rate of OHSS was much lower in the 10∼12 group (11.53 %) than that in the 13∼15 group (29.97 %) and ≥16 group (77.30 %). Unconditional multivariate logistic regression analysis showed that when ≥10 oocytes were retrieved, the CLBR increased significantly (P < 0.01). When oocyte number exceeded 16, the CPR of frozen embryo transfer cycle was much higher than that of fresh cycle (P < 0.05).

Conclusions

For young women with normal ovarian reserve, retrieving 10∼12 oocytes might result in optimized pregnancy outcomes in a fresh cycle with low OHSS risk and would not compromise cumulative outcomes. When ≥16 oocytes were retrieved, a “freeze-all” embryo strategy might be preferable.     

Does company-sponsored egg freezing promote or confine women’s reproductive autonomy?

Purpose

A critical ethical analysis of the initiative of several companies to cover the costs of oocyte cryopreservation for their healthy employees. The main research question is whether such policies promote or confine women’s reproductive autonomy.

Results

A distinction needs to be made between the ethics of AGE banking in itself and the ethics of employers offering it to their employees. Although the utility of the former is expected to be low, there are few persuasive arguments to deny access to oocyte cryopreservation to women who are well informed about the procedure and the success rates. However, it does not automatically follow that it would be ethically unproblematic for employers to offer egg banking to their employees.

Conclusions

For these policies to be truly ‘liberating’, a substantial number of conditions need to be fulfilled, which can be reduced to three categories: (1) women should understand the benefits, risks and limitations, (2) women should feel no pressure to take up the offer; (3) the offer should have no negative effect on other family-friendly policies and should in fact be accompanied by such policies. Fulfilling these conditions may turn out to be impossible. Thus, regardless of companies’ possible good intentions, women’s reproductive autonomy is not well served by offering them company-sponsored AGE banking.     

Laparoscopic excision of ovarian endometrioma does not exert a qualitative effect on ovarian function: insights from in vitro fertilization and single embryo transfer cycles

Purpose

To evaluate whether laparoscopic excision of endometrioma exerts a qualitative effect on ovarian function.

Methods

A retrospective analysis of oocytes retrieved in 25 cycles of 21 patients undergoing IVF treatment with controlled ovarian stimulation. The number of oocytes recovered from ovaries with a history of excision of endometrioma (E-Ov) were compared to those from contra-lateral healthy ovaries (H-Ov) as for the analysis of a quantitative effect of surgery. As for the analysis of a qualitative effect, 55 oocytes from E-Ov were compared to 128 oocytes from H-Ov in terms of normal fertilization rate and the rate of top-quality embryos per normally fertilized eggs. Furthermore, 10 embryos derived from oocytes recovered from E-Ov were compared to 24 embryos derived from oocytes from H-Ov in terms of clinical and on-going pregnancy rates per embryos in 34 single embryo transfer cycles.

Results

Mean number of oocytes recovered from E-Ov was significantly smaller than that from H-Ov (2.2 ± 2.0 vs. 5.1 ± 3.3, P = 0.009). There was no difference between oocytes from E-Ov and H-Ov as for normal fertilization rate (63.6 % vs. 69.5 %, P = 0.43) and the rate of top-quality embryos (40.0 % vs. 49.0 %, P = 0.34). Clinical and on-going pregnancy rates per embryos were also similar in embryos derived from oocytes recovered from E-Ov and H-Ov (40.0 % vs. 25.0 %, P = 0.39 and 20.0 % vs. 20.8 %, P = 0.96).

Conclusions

The quality of oocytes recovered from the ovary with a history of laparoscopic excision of endometrioma is not inferior to the quality of oocytes from contra-lateral healthy ovary.     

Live birth following IVF/ICSI using oocytes from donor who was conceived via IVF: a case report

Purpose

The purpose of the study was to report a case of live birth following donor oocyte in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) in which the oocyte donor herself was conceived via IVF. To our knowledge, such a case has not been previously reported.

Methods

Retrospective chart review; this case is reported after chart review of a successful outcome.

Results

A 42 year-old woman, with diminished ovarian reserve, and her husband desired to conceive. She underwent a fresh IVF/ICSI cycle with her own oocytes, which unfortunately was not fruitful in terms of pregnancy or cryopreserved embryos. The couple was counseled regarding the option of donor oocytes, and they elected to proceed with a fresh cycle of donor oocyte IVF/ICSI. The couple selected an anonymous oocyte donor from a donor agency who was a first-time oocyte donor and, interestingly, was conceived via IVF herself. The fresh donor oocyte/IVF/ICSI cycle did not result in pregnancy; however, two supernumerary blastocysts were cryopreserved for future cycles. The recipient’s subsequent frozen-thawed embryo transfer (FET) resulted in a singleton gestation and live birth.

Conclusions

An oocyte donor who was conceived via IVF had good ovarian response to stimulation, a good number of oocytes retrieved, and the formation and cryopreservation of blastocysts which, in a subsequent FET cycle, resulted in pregnancy and live birth for a recipient couple. To our knowledge, this is the first case reported of live birth with the use of donor oocytes from an oocyte donor who herself was conceived via IVF.     

小鼠卵子老化过程中皮质颗粒的变化和自发孤雌激活的研究

目的:通过卵子老化过程中皮质颗粒的变化和自发孤雌激活比例,探讨老化卵子受精异常的原因。方法:对8～10周龄雌鼠注射孕马血清促性腺激素和绒毛膜促性腺激素14h后,于不同时间段取出MII期成熟卵子分作体内不同时间老化组(MII+0h、MII+6h、MII+12h、MII+18h);成熟MII+0h卵子取出后体外分别培养6h、12h、18h作为体外老化组。对各组卵子免疫荧光染色,共聚焦显微镜成像显示染色体形态和皮质颗粒分布。体内各组卵子取出后继续体外培养8h统计自发孤雌激活的比例和碎裂卵子数。结果:体内和体外MII+12h、MII+18h组自发皮质颗粒排放比例和皮质颗粒内迁数明显高于MII+0h、MII+6h组。体外MII+18h组自发皮质颗粒排放比例和皮质颗粒内迁数低于体内MII+18h组。体内MII+12h、MII+18h组卵子取出继续体外培养8h后孤雌激活比例和死亡比例明显高于体内MII+0h和MII+6h组。结论:卵子在老化过程中存在皮质颗粒自发排放和皮质颗粒内迁,以及自发孤雌激活和碎裂化,这些因素均可能导致老化卵受精能力的降低。卵子老化过程中皮质颗粒排放和内迁可能受到体内环境的影响。