长方案超排卵获卵数对胞浆内单精子注射治疗结局的影响

目的：比较常规长方案超排卵周期中,获取不同卵母细胞数对胞浆内单精子注射（ICSI）治疗结局的影响。方法：回顾分析广州医科大学附属第三医院生殖医学中心2007年1月—2013年6月因单纯男性因素采用常规长方案超排卵行ICSI治疗的患者资料。根据获卵数目分成4组,获卵数≤4个为G1组（90个周期）,获卵数5~10个为G2组（368个周期）,获卵数11～20个为G3组（449个周期）,获卵数≥21个为G4组（135个周期）。结果：在ICSI治疗中,G1组女方平均年龄高于其他3组（P＜0.01）,但其中＜35岁的患者占57.8%。G1组患者完全受精失败发生率、G4组的卵巢过度刺激综合征（OHSS）发生率以及这2组的移植周期取消率均高于G2组和G3组（P＜0.01）;G1组的临床妊娠率在4组中最低（31.3%,P＜0.05）;4组中G3组的临床妊娠率、胚胎种植率和移植胚胎活产率最高,且后两者差异有统计学意义（P＜0.05）。结论：在ICSI治疗周期中,获卵数≤4个影响ICSI受精结局,完全受精失败发生率升高;获卵数在11～20个时ICSI临床结局最好。     

对常规IVF授精后4小时未排出第二极体的卵母细胞行补救ICSI

目的比较常规IVF授精后4h和20h补行卵胞浆内单精子注射（rescue ICSI）的效果。方法回顾分析2006年1月-2007年12月在我院生殖医学中心接受常规IVF治疗的21例非男性因素不育患者,授精后4h行补救ICSI,并与另组实施的18个常规IVF授精后20h行补救ICSI周期进行比较。结果 4h补救ICSI组的受精率为76.3%,高于20h补救ICSI组（53.0%）,有统计学意义（P〈0.05）。4h补救ICSI组中11例临床妊娠（52.3%）,其中2个双胎,8个单胎和一个流产。而20h补救ICSI组无一例妊娠。结论和授精后20h行补救ICSI相比,授精后4h行补救ICSI可以获得较好的受精率、妊娠率和种植率。     

The effect of minimal concentration of ethylene glycol (EG) combined with polyvinylpyrrolidone (PVP) on mouse oocyte survival and subsequent embryonic development following vitrification

Purpose

Vitrification techniques employ a relatively high concentration of cryoprotectant in vitrification solutions. Exposure of oocytes to high concentrations of cryoprotectant is known to damage the oocytes via both cytotoxic and osmotic effects. Therefore, the key to successful vitrification of oocytes is to strike a balance between the usage of minimal concentration of cryoprotectant without compromising their cryoprotective actions.

Methods

The minimal concentration of ethylene glycol (EG) on mouse oocyte survival and subsequent embryonic development was evaluated following vitrification-warming and parthenogenetic activation. Polyvinylpyrrolidone (PVP) combined with EG on mouse oocyte survival and subsequent embryonic development as well as morphology of the spindle and chromosome alignment were also evaluated. Vitrification system was adapted with JY Straw and the cooling rate was approximately 442–500 °C/min. In contrast, the warming rate was approximately 2,210–2,652 °C/min.

Results

Survival rate of oocytes increased significantly when 15 % EG was combined with 2 % PVP in vitrification solution (VS). The effect of combination of EG and PVP was not significant when the concentration of EG was 20 % and higher. Although there were no significant differences in embryonic development, the percentage of abnormal spindle and chromosome alignment was significantly higher in the oocytes without 2 % PVP in VS.

Conclusions

Our data provide a proof of principle for oocyte vitrification that may not require a high concentration of cryoprotectant. There are synergic effects of EG combined with PVP for oocyte vitrification, which may provide important information to the field in developing less cytotoxic VS.     

Protein phospholipase C Zeta1 expression in patients with failed ICSI but with normal sperm parameters

Purpose

This study was conducted to determine if expression of the testis-specific phospholipase C Zeta1 (PLCZ1) correlated with low success or fertilization failure after ICSI in patients with normal parameters after standard semen analysis (SA).

Methods

Couples <43 years with one or two failed or low fertilization ICSI cycles. Standard Semen Analysis (SA) was performed to determine sperm parameters in male partners, whereas females were evaluated for antral follicle counts (AFC), day 3 FSH levels and peak Estradiol (E2) levels. The presence of PLCZ1 in sperm was ascertained using Western blotting and Immunofluorescence (IF) analysis. The ability of sperm to initiate changes in the intracellular concentrations of free calcium ([Ca²⁺]_i), which is characteristic of mammalian sperm, was performed after injection of human sperm into mouse eggs loaded with the Ca²⁺ sensitive dye fura-2 AM.

Results

Male partners of couples with failed or low success ICSI fertilization but with normal SA parameters showed low expression levels of PLCZ1 as determined by western blotting and reduced fluorescent signal during IF studies. In addition, fewer of these males’ sperm showed PLCZ1 expression and were able to initiate robust [Ca²⁺]_i oscillations upon injection into eggs.

Conclusion

Our data suggest that in patients with normal SA parameters but with repeated low fertilization or outright failed fertilization results after ICSI, abnormal PLCZ1 function should be considered as the underlying mechanism responsible for the failure of fertilization.     

IVF周期中MⅡ末期卵子形态与胚胎发育潜能的关系

目的:探讨IVF周期中MⅡ末期卵子形态与胚胎发育潜能的关系。方法:收集IVF周期中受精后5～6h脱颗粒细胞MⅡ末期卵子,依据透明带、卵泡浆及两极体形态在光镜下对排出两个极体的卵子进行形态评分分级(Ⅰ～Ⅲ级),统计不同分级组间的高评分原核形成率、卵裂率、优质胚胎率及临床妊娠率和种植率,分析卵子形态与受精及胚胎发育潜能的关系。结果:形态评分较高的卵子与形态评分较低的卵子间受精率和卵裂率均无显著差异(P>0.05),但优质胚胎率显著增高(P<0.05);随着移植胚胎中由高形态评分卵子发育而来的胚胎数目增多,周期临床妊娠率、种植率显著增高(P<0.05)。结论:IVF周期中,早期脱颗粒细胞后MⅡ末期卵子形态对受精率及卵裂率没有影响,但可预测早期胚胎质量及妊娠结局。     

Cumulus cell-oocyte complexes retrieved from antral follicles in IVM cycles: relationship between COCs morphology, gonadotropin priming and clinical outcome

Purpose

To assess retrospectively the developmental potential of different types of cumulus cell-oocyte complexes (COCs) derived from IVM cycles.

Methods

IVM cycles were performed in natural cycles or after HCG, FSH, or FSH/HCG priming. COCs recovered were morphologically characterized in different types: compact (CC) or expanded (EC) cumulus mass but including an immature oocyte, and expanded cumulus mass enclosing a mature oocyte (EC-MII). Embryo developmental competence was investigated analysing exclusively cycles in which all transferred embryos derived from the same COC category.

Results

Fertilization rates did not differ significantly. Significant differences in pregnancy rates (14.5 %, 10.0 % and 27.6 % in the CC, EC, and EC-MII categories, respectively) were observed. Likewise, significant differences in implantation rates (8.9 %, 6.3 % and 19.1 % in the CC, EC, and EC-MII categories, respectively) were found. Overall, priming with FSH/HCG had a beneficial effect on pregnancy and implantation rates, while no priming or HCG alone generated oocytes with poor competence.

Conclusions

In IVM cycles, morphological evaluation at the time of collection can predict the developmental ability of different COCs. FSH/HGC priming has a positive effect on oocyte competence.     

Rare live birth to a 48-year-old woman after embryo transfer with autologous oocyte: A case report

ObjectiveAdvanced maternal age and decreased ovarian reserve have been challenges for assisted reproductive technology (ART). Few cases, using autologous oocytes more than 46-years-old, have previously been reported. We seek to show how the age at which autologous oocytes may successfully be employed may be increasing.Case reportWe report a 47-year-old woman with an anti-Müllerian hormone (AMH) level of 0.24 ng/mL, conceived through in vitro fertilization (IVF) using autologous oocytes. Patient was given an antagonist protocol for ovarian stimulation and one frozen-thawed embryo was transferred. The patient became pregnant. The course of her pregnancy was uneventful and she gave birth to a 3330 gm male baby by cesarean section.ConclusionTechnological advances permit women, who previously would have been considered too old to employ an autologous oocyte, to have a successful pregnancy with a live birth.