囊胚发育速度及质量对冻融周期单囊胚选择的影响

目的联合囊胚发育速度和囊胚质量分析其对妊娠结局和新生儿出生结局的影响,为冻融周期中的囊胚选择提供证据。方法回顾性分析2011年7月至2018年6月在北京协和医院生殖中心进行单囊胚复苏移植治疗的患者资料,周期数为736个。按照囊胚形成时间分为两组:D5组(在受精第5天形成囊胚)和D6组(在受精后第6天形成囊胚)。再根据各组内囊胚是否为优质胚胎分为4组,分别为D5优胚组、D5非优胚组,D6优胚组、D6非优胚组。分别比较各组的临床妊娠结局和新生儿的出生结局。结果 (1)共纳入736个复苏周期,复苏存活率100%。患者平均胚胎复苏年龄为(35.1±4.4)岁。临床妊娠率为40.9%,流产率为19.6%,活产率为26.0%。(2)D5组中移植的优质胚胎比例显著高于D6组(60.4%vs. 36.6%,P<0.05);D5组的临床妊娠率和活产率均显著高于D6组(分别为51.5%vs. 33.6%和33.9%vs. 20.6%)(P均<0.05);两组的流产率无显著差异(P>0.05)。D5组的男女性别比略高于D6组(P>0.05)。两组新生儿的身长和出生体重无显著差别(P>0.05)。D5组巨大儿的发生率略高于D6组,但无显著差异(P>0.05)。两组新生儿均未发生出生死亡和先天异常。(3)D5优胚组、D5非优胚组、D6优胚组和D6非优质胚胎组的临床妊娠率和活产率均有显著性差异(P均<0.05),D5优胚组的临床妊娠率和活产率最高,D6非优胚组的临床妊娠率和活产率最低。各组间的新生儿出生体重、身长和男女性别比无统计学差异(P>0.05)。D5非优胚组的临床妊娠率和活产率均略低于D6优胚组,但两者均无显著差异(P>0.05)。D6非优胚组的临床妊娠率显著低于D5非优胚组(P<0.05),但两组的活产率无显著差异(P>0.05)。结论单囊胚的冻融周期移植时,优先选择D5的优质胚胎;而面临D5非优质胚胎和D6优质胚胎时,两者均可考虑,可以根据实验室的实际情况进行选择。     

Melatonin promotes the in vitro development of pronuclear embryos and increases the efficiency of blastocyst implantation in murine

When a defect occurs in the in vitro development of a pronuclear embryo, the interruption of the subsequent implantation limits the success of assisted conception. This common problem remains to be solved. In this study, we observed that melatonin at its physiological concentration (10^?7 m ) significantly promoted the in vitro development of murine pronuclear embryos. This was indicated by the increased blastocyst rate, hatching blastocyst rate, and blastocyst cell number with melatonin treatment. In addition, when these blastocysts were implanted into female recipient mice, the pregnancy rates (95.0% versus control 67.8%), litter sizes (4.1 pups/litter versus control 2.7 pups/litter), and postnatal survival rates of offspring (96.84% versus control 81.24%) were significantly improved compared with their non‐melatonin‐treated counterparts. Mechanistic studies revealed that melatonin treatment upregulates gene expression of the antioxidant enzyme, superoxide dismutase (SOD), and the anti‐apoptotic factor bcl‐2 while downregulating the expression of pro‐apoptotic genes p53 and caspase‐3. Due to these changes, melatonin treatment reduces ROS production and cellular apoptosis during in vitro embryo development and improves the quality of blastocysts. The implantation of blastocysts with higher quality leads to more healthy offspring and increased pup survival.     

“Pinopodes” and Implantation

Reviews in Endocrine and Metabolic Disorders -     

复苏周期单囊胚移植的影响因素分析

目的研究玻璃化复苏囊胚移植后的妊娠结局与冷冻前囊胚质量、发育速度的相关性,探讨单囊胚移植的可能性。方法回顾分析105例复苏周期单囊胚移植患者的临床资料,观察发育速度、囊胚质量、内细胞团(ICM)/滋养外胚层(TE)评分对妊娠结局的影响。结果第5、6天囊胚间复苏后存活率、移植后妊娠率的差异均无统计学意义(98.0%,54.5%vs 97.0%,52.5%)。优质囊胚移植后妊娠率(44/70,62.9%)显著高于非优质囊胚(12/35,34.3%)。ICM评分为A、B级的囊胚移植后妊娠率差异无统计学意义(70.6%vs 58.0%),但均显著高于C级胚胎(21.1%)。TE评分越低,妊娠率有下降趋势但差异无统计学意义。结论复苏周期单囊胚移植妊娠率与囊胚质量有关,与囊胚发育速度无关。ICM评分而非TE评分与妊娠结果显著相关。复苏周期单个优质囊胚移植后妊娠率高,提示单囊胚移植可行。     

胚胎移植与囊胚移植的临床结局分析*

背景：国内关于卵裂期胚胎移植与囊胚移植的临床比较与分析大样本报道较少见。 目的：比较第2,3天卵裂期胚胎移植与第5天囊胚移植的临床结局。 方法：选择2008-01/2009-12北京大学深圳医院生殖医学科行体外受精/卵胞浆内单精子显微注射共1612周期,比较第2,3天卵裂期胚胎移植与第5天囊胚移植的临床妊娠率、胚胎种植率、流产率、多胎率、出生婴儿性别比例等差异。 结果与结论：第2,3,5天移植组分别195,1 162,255周期,临床妊娠率分别为33.33%,38.04%,44.71%,种植率分别为21.37%,24.70%,31.96%;第5天移植组临床妊娠率和种植率均高于第2,3天移植组(P < 0.05);第2天移植组宫外孕发生率高于第3天移植组(P < 0.05)。3组流产、单卵双胎、多胎、早产及出生婴儿性别比例无差异。说明囊胚移植提高了临床妊娠率和种植率的同时,未增加流产、多胎、早产等风险,对男女性别比例的平衡无负面影响。     

囊胚玻璃化冻存前激光打孔对冻融后移植效果的影响

背景：一系列研究均表明人工皱缩囊胚腔能明显提高囊胚的玻璃化冻融效果,但在皱缩过程中采用的物理方法和使用材料的不同可能会造成囊胚细胞的机械性损伤。 目的：探讨囊胚玻璃化冻存前激光打孔对冻融后移植效果的影响。 方法：3例不孕患者,平均年龄30岁,于囊胚玻璃化冻存前行激光打孔使囊胚皱缩,平均冻存时间为1年,确定移植前给予解冻。 结果与结论：3例患者中有2例取得了满意的妊娠结局,1例未妊娠。说明采用激光辅助打孔的方法使囊胚皱缩,结合玻璃化冻融后行囊胚移植安全有效。     

Self Renewal of Embryonic Stem Cells in the Absence of Feeder Cells and Exogenous Leukaemia Inhibitory Factor

Abstract

To evaluate the role of leukaemia inhibitory factor (LIF) for maintaining pluripotent embryonic stem (ES) cells in culture, we established several exogenous LIF-independent ES cell lines by continuous passaging in culture. The newly established ES cells, Kli and CBli, sustained their growth and remained undifferentiated in LIF-deficient medium. Analysis of chimaeric animals, produced with the p-galactosidase transgenic Kli ES cells, revealed that LDF-independent ES cells can contribute to all embryonic germ layers. There was no detectable LIF protein in ES cell conditioned medium, and no upregulation of LIF mRNA was found. The addition of neutralising anti-LIF antibodies was not sufficient to abrogate the self renewal of the Kli ES cells. These studies suggest that the signalling pathway involving diffusible LIF can be bypassed for maintaining the pluripotency in culture, and indicate a considerable heterogeneity in growth factor dependence and differentiation of different ES cells.     

激光削薄透明带对小鼠囊胚形成和孵出的影响

目的探索激光削薄透明带对体外培养昆明小鼠囊胚形成和孵出的影响。方法激光孵化组为应用1．48μm二极管激光对186枚2细胞期昆明小白鼠胚胎实施透明带削薄处理;对照组不削薄透明带,在离胚胎一个直径处发射激光。比较两组囊胚形成率、孵出率、完全孵出率、透明带厚度和胚胎直径。结果激光组和对照组的囊胚形成率分别为73．7％（137／186）和71．6％（53／74）,两组比较差异无统计学意义。激光组的孵出率（86．9％,119／137）明显高于对照组（71．7％,38／53,P〈0．05）;囊胚嵌顿率（74．8％,89／119）明显高于对照组（23．1％,9／38,P〈0．001）。结论激光辅助孵化对体外培养昆明小鼠囊胚形成无不良影响且可以提高囊胚孵出率,但囊胚嵌顿率显著升高。