冻融胚胎移植30例临床分析

目的探讨冻融胚胎移植(F-ET)治疗的冷冻、融解及妊娠情况。方法对2005年接受常规IVF-ET治疗不孕症患者后剩余胚胎冻融移植33周期临床资料进行回顾性分析。结果33周期中共冷冻胚胎190个,融解胚胎157个,移植97个,融胎存活率79.62%,临床妊娠率33.33%,种植率13.40%。结论F-ET是辅助生育技术治疗不孕症的重要组成部分,采用冻融胚胎技术可提高一次取卵的总妊娠率。冻融胚胎质量和有足够移植胚胎数是成功的关键;冻融胚胎移植时机是冻融胚胎移植成功的重要因素。     

不同发育时期的冻融胚胎移植的成功率分析

目的探讨辅助生殖技术中,胚胎冷冻时的发育阶段对冻融胚胎移植的成功率的影响.方法回顾性分析107个冻融胚胎移植周期.结果胚胎复苏率为57.2%(271/474),临床妊娠率为27.1% (29/107),种植率为15.1% (39/259).按胚胎冷冻时期不同分为受精后第二天组(2～6细胞)和第三天组(4～10细胞),Day 3组的胚胎存活率显著高于Day 2组,但完整胚胎存活率、妊娠率、种植率均无显著性差异(P＞0.05).妊娠组的完整胚胎存活率及移植胚胎数目明显高于未妊娠组(P＜0.05).两组的子宫内膜的厚度和分型差异无显著性.结论在受精后第二天或第三天冷冻的胚胎,复苏后胚胎移植的妊娠率、种植率均无显著性差异.冻融胚胎形态良好,有足够的移植胚胎数目是保证冻融胚胎移植成功的重要因素.     

血LH峰与尿LH峰测定判断冻融胚胎移植时机的方法学比较

目的研究利用电化学发光免疫分析技术监测血LH板或利用尿LH板监测尿LH峰判断冻融胚胎移植时机的方法学比较。方法本中心由1999年5月至今共完成冻融胚胎移植67例,排除冻融胚胎移植周期中所移植的胚胎全部为部分卵裂球受损胚胎的周期后,将其中61例移植周期随机分成A、B两组,A组共12周期,通过使用尿LH板测定尿LH峰来判断冻融胚胎移植时机,B组共49个周期,通过电化学发光免疫分析技术测定血LH峰来判断冻融胚胎移植时机,对两组资料进行回顾性分析。结果两组患者平均冻融胚胎移植数目、冷冻前胚胎形态学评分及解冻后胚胎卵裂球存活程度等均无显著差异(P>0.05),冻融胚胎移植周期的子宫内膜厚度均大于8mm。利用尿LH峰测定判断冻融胚胎移植时机的12周期中,移植冻融胚胎37个,平均每周期移植胚胎3.1±0.4个,周期临床妊娠率33.3%(4/12)。利用血LH峰测定判断冻融胚胎移植时机行冻融胚胎移植的46例49个周期中,移植冻融胚胎137个,平均每周期移植胚胎2.8±0.1个,周期临床妊娠率63.3%(31/49),两组资料临床妊娠率有非常显著性差异(P<0.01)。结论利用电化学发光免疫分析技术监测血LH峰判断冻融胚胎移植时机比利用尿LH板监测尿LH峰判断冻融胚胎移植时机更为准确和灵敏,显著提高临床妊娠率。     

冷冻时间对冻融胚胎移植周期结局的影响

目的探讨受精后第二天（D2）和第三天（D3）人胚胎冷冻对冻融胚胎移植周期结局的影响。方法回顾性分析本中心403个D2和D3胚胎程序化慢速冷冻-快速解冻周期,冷冻复苏后的胚胎存活率、完整率、种植率、临床妊娠率等指标。结果 D2冻融移植周期291个,存活率98.2%,完整率78.5%,胚胎种植率16.9%,临床妊娠率33.0%;D3解冻移植周期112个,存活率95.8%,完整率67.5%,胚胎种植率21.5%,临床妊娠率34.8%。D2比D3能显著提高胚胎存活率和完整率（P〈0.05）,但在临床妊娠率、胚胎种植率上无显著性差异。D2和D3胚胎在当天和提前解冻组的临床妊娠率、胚胎种植率差异没有显著意义。结论 D3胚胎冷冻没有比D2胚胎更有优势,D2胚胎对冷冻解冻可能具有更好的耐受。可以根据工作需要以及病人情况选择冷冻时间。     

提前复苏分裂速率较慢胚胎对妊娠率的影响

目的探讨冻融胚胎移植(FET)周期中,提前一日复苏冷冻前分裂速率较慢的胚胎对妊娠结局的影响。方法回顾性分析哈尔滨医科大学附属第一医院生殖中心2006年3月至2009年9月561例FET周期中,移植冷冻前分裂速率较慢胚胎的周期共88个。其中,33个周期为移植当日复苏胚胎,培养1-4h后移植。55个周期为移植前一日复苏胚胎,培养18-24h移植。比较两组胚胎移植后的妊娠率和种植率。结果移植当日复苏组与移植前一日复苏组妊娠率(9.1%,32.7%)与种植率(4.2%,15.5%)均有显著性差异(P﹤0.05);而患者平均年龄、移植前平均内膜厚度、胚胎复苏存活率及100%卵裂球存活率均无显著性差异(P﹥0.05)。结论对冷冻前分裂速率较慢胚胎提前一日复苏,可提高临床妊娠率和种植率。     

冻融胚胎和冻融囊胚对移植周期和分娩结局影响的比较

背景：自从1983年人类首例冷冻胚胎移植取得成功以来,胚胎冷冻技术已成为人类辅助生殖技术中重要组成部分。对选择冻融胚胎还是选择冻融囊胚移植,各地都有不少争议。 目的：比较解胚胎和囊胚经过低温保存解冻复苏后的分娩结局及新生儿状况。 方法：比较冻融胚胎移植周期1 273例和冷冻囊胚移植周期471例两组妊娠率、流产率、异位妊娠率、早产率、平均早产孕周、足月产率、平均足月产孕周、新生儿男女性别比例、出生体质量、出生缺陷等指标。 结果与结论：冻融囊胚解冻周期478例,移植周期471例(其中7例无囊胚移植取消),妊娠236例,分娩201例,分娩胎数251胎,男孩140个,女孩111个。冻融第3天胚胎解冻周期1 280例,移植周期1 273例(其中7例无胚胎移植取消),妊娠415例,分娩343例,分娩胎数431胎,男孩225个,女孩206个。冻融囊胚的妊娠率显著高于冻融胚胎妊娠率。冻融胚胎和囊胚的流产率、异位妊娠率、早产率、平均早产孕周、足月产率、平均足月产孕周、新生儿男女性别比例、出生体质量等差异无显著性意义。冻融胚胎和冻融囊胚移植出生缺陷并未明显增加。结果表明冻融囊胚与冻融胚胎的分娩结局及新生儿状况差异无显著性意义,但冻融囊胚的妊娠结局优于冻融胚胎的妊娠结局。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

高龄妇女新鲜胚胎和冻融胚胎移植的结局比较

目的探讨高龄妇女接受新鲜胚胎和冻融胚胎移植后的结局。方法对2008年1月～2010年12月期间在我院接受体外受精-胚胎移植和玻璃化冻融胚胎移植的36岁以上的高龄妇女的临床资料进行回顾性分析。结果新鲜胚胎移植306例,383个周期,90个周期获得妊娠,妊娠率23.5%,无畸形胎儿;玻璃化冻融胚胎移植75例,94个周期,27个周期获得妊娠,妊娠率28.72%,1例畸胎（脑积水）,行中孕引产。两种胚胎移植的妊娠率、多胎率、胚胎种植率、早期流产率、异位妊娠率等比较,差异均无统计学意义（P〉0.05）。结论高龄妇女接受新鲜胚胎移植和冻融胚胎移植的妊娠率、多胎率、胚胎种植率、早期流产率、异位妊娠率无显著性差异。由于高龄妇女卵巢和内膜功能下降,建议应在有限的时间内增加胚胎移植的次数来提高累计妊娠率。     

体外受精-胚胎移植的助孕效果:胚胎体外培养和低温保存

背景:从第一例试管婴儿诞生至今已二十余年,体外受精-胚胎移植技术在世界范围内得到蓬勃发展。目的:回顾性分析体外受精-胚胎移植的助孕效果,探讨体外受精胚胎的培养及低温保存技术的应用。方法:2013年泸州医学院附属医院生殖医学技术部接受体外受精-胚胎移植或卵细胞浆内单精子注射助孕的不孕患者131例,共147个取卵周期,124个移植周期,患者平均年龄31.32岁。有131个取卵周期应用标准长方案促排卵,4个取卵周期应用超长方案促排卵、10个取卵周期应用超短方案促排卵,2个取卵周期为自然周期排卵;应用新鲜胚胎移植93例,冷冻胚胎移植28例。观察不同促排卵方案、新鲜或冷冻胚胎移植患者的临床妊娠率、胚胎植入率和流产率等。结果与结论:应用标准长方案、超长方案及超短方案促排卵患者的临床妊娠率为34.7%,50%,20%;植入率分别为19.1%,22.2%,30%;流产率为21.7%,50%,33.3%;以上指标自然周期排卵患者均为0。新鲜胚胎移植临床妊娠率33.6%,植入率18.1%,流产率20.1%;冷冻胚胎移植临床妊娠率53.8%,植入率22.3%,流产率20.7%。提示应用标准长方案促排卵效果好,新鲜及冷冻胚胎移植的疗效相当,胚胎的体外培养及低温冷冻是辅助生殖技术中重要步骤,需进一步深入研究。     

冻融胚胎移植临床结局的回顾性分析

目的探讨影响冻融胚胎移植(frozen-thawed embryo transfer,FET)妊娠结局的相关因素。方法回顾性分析本中心132例冻融胚胎移植周期,其中1例取消移植外,将另外131例移植周期按照妊娠结局分为妊娠组和未妊娠组进行研究。结果 132周期共解冻胚胎290枚,存活280枚(96.55%),移植131周期,临床妊娠率为48.85%(64/131),胚胎着床率为30.00%。患者的基本资料之间无显著性差异,FET周期临床资料中,妊娠组患者的胚胎复苏率、完全存活率、冷冻前和复苏后的优胚率、继续发育胚胎比例及移植胚胎中含有致密化以上胚胎比例均显著高于未妊娠组(99.26%vs94.77%;92.65%vs 77.78%;94.12%vs 84.31%;91.11%vs 77.93%;68.89%vs 39.31%;40.74%vs 18.62%)。FET周期患者在新鲜取卵周期中,两组的Gn用量与天数、获卵数、受精率及新鲜周期胚胎移植失败率之间无显著性差异,但妊娠组的优胚率和冷冻胚胎数均显著高于未妊娠组(65.80%vs 55.72%;7.69 vs 5.03)。结论在冻融胚胎移植中,患者新鲜周期的胚胎质量、胚胎冻融过程以及解冻后的胚胎发育情况共同决定其妊娠结局     

冻融囊胚及冻融胚胎养囊胚后移植的临床效果分析

目的探讨新鲜胚胎行囊胚培养后冷冻,再经囊胚复苏移植与冻融胚胎复苏后再行囊胚培养形成囊胚后移植的效率比较。方法解冻后有2个以上的II级胚胎或者Ⅲ级胚胎超过4个者行囊胚培养,形成3CC以上囊胚则进行移植,同期将新鲜卵裂期胚胎移植或冷冻部分优胚后剩余的胚胎进行囊胚培养,形成3CC以上的囊胚进行冷冻,再次复苏囊胚移植。结果冻融胚胎解冻行囊胚培养共139个周期,解冻存活694个胚胎,形成3CC以上囊胚228个,囊胚形成率32.85%,有囊胚形成的119个周期,移植118个周期,移植妊娠率和种植率分别为41.5%和30.2%,同期进行新鲜剩余胚胎的囊胚培养141个周期共1204个胚胎,形成3CC以上的囊胚386个,有囊胚形成的128个周期进行冷冻,解冻移植38个周期,移植妊娠率和种植率分别为50.0%和35.7%。结果解冻囊胚移植的妊娠率与种植率比解冻后养囊胚获移植的妊娠率与种植率高,但两组无统计学差异。新鲜胚胎养囊胚和冻融胚胎复苏后养囊胚的形成率和取消率没有统计学差异。结论经历多次优质胚胎移植后妊娠失败的患者可考虑将复苏胚胎行囊胚培养后移植囊胚来改善妊娠结局;在新鲜周期移植冷冻部分优胚后剩余的胚胎行囊胚培养后再冷冻,可提高胚胎的有效利用率。     

Cryo-thawed embryos obtained from conception cycles have double the implantation and pregnancy potential of those from unsuccessful cycles

BACKGROUND: The purpose of this study was to evaluate the influence of fresh IVF/ICSI cycle outcome on the prognosis of the related frozen embryo replacement (FER) cycle. METHODS: 459 FER cycles, involving 2049 cleavage stage embryos with no or up to 10% fragmentation, were performed for which the outcome of the fresh cycle was recorded. The cycles were divided into two groups; group A included cycles in which cryopreserved embryos were obtained from fresh cycles in which conception occurred. Group B were cycles in which cryopreserved embryos originated from unsuccessful fresh cycles. RESULTS: Groups A and B were comparable with respect to mean (+/- SD) age at cryopreservation (33 +/- 3.9 versus 33.2 +/- 4 years, P = not significant), mean number of oocytes retrieved and fertilized normally in the fresh cycle (11 +/- 5.2 versus 11.2 +/- 4.8, P = not significant) and mean age at the cryo-thawed transfer (34.5 +/- 4.2 versus 33.9 +/- 4 years, P = not significant). No significant difference was found between the two groups with regard to mean number of embryos cryopreserved (6.5 +/- 3.9 versus 6.2 +/- 3.6) and subsequently thawed (4.5 +/- 2.5 versus 4.5 +/- 1.8) per cycle and number of cryo-thawed embryos transferred per cycle (2.0 +/- 0.7 versus 2.1 +/- 0.8). However, the implantation rate per transferred embryo in group A was double that in group B (23 versus 11.2%, P < 0.0001). Moreover, the clinical pregnancy and ongoing pregnancy rates per cycle were significantly higher in group A compared with group B (34.8 and 27.3% versus 15.6 and 13.1%, P < 0.0001 and P = 0.0003 respectively). The difference in FER cycle outcome could not be explained by confounding variables. CONCLUSIONS: After thawing, cryopreserved embryos originating from conception IVF/ICSI cycles achieve double the implantation and pregnancy rates of those obtained from unsuccessful cycles.     

The embryo versus endometrium controversy revisited as it relates to predicting pregnancy outcome in in-vitro fertilization-embryo transfer cycles

To evaluate embryonic and endometrial factors for their value in predicting pregnancy outcome in in-vitro fertilization (IVF) and embryo transfer, a retrospective data collection and prospective uterine artery colour Doppler imaging study was performed in a university-based IVF-embryo transfer programme setting. A total of 210 patients were included and grouped as follows: (I) IVF with controlled ovarian stimulation (214 cycles); (II) frozen-thaw cycle of autologous embryos (30 cycles); (III) oocyte donation, no cryopreservation (12 cycles); (IV) frozen-thaw cycle with embryos from donated oocytes (10 cycles). Embryo quality was significantly better in pregnant than non-pregnant cycles (group I, P = 0.0104; groups II-IV, P = 0.0418). The endometrial echo was significantly thicker in pregnant versus non-pregnant patients in group I (P = 0.0059), but not in groups II-IV (P = 0.741). Past uterine surgery or abnormalities had no effect on pregnancy outcome. There were no significant differences in mean uterine artery resistance index or peak systolic velocity in pregnant versus non-pregnant patients in groups II-IV. Thus, embryo quality is the most reliable predictor of pregnancy outcome. Endometrial measurements were significantly thicker in subsequently pregnant patients only in group I, where the endometrium reflects the hormonal environment. Doppler parameters were not useful in predicting pregnancy outcome.      

Reduced pregnancy rates following the transfer of human embryos frozen or thawed in culture media supplemented with normal serum albumin

Over a 26 month period 17% of couples having treatment in our clinical programmes selected a commercially available protein (normal serum albumin, NSA) prepared from pooled human sera instead of using their own serum as a supplement for their embryo culture media. In a retrospective analysis of >2000 gonadotrophin-stimulated cycles and 1000 cycles where frozen/thawed embryos were transferred, fertilization, embryo quality and pregnancy rates following in-vitro fertilization (IVF), gamete intra-Fallopian transfer (GIFT) or intracytoplasmic sperm injection (ICSI) were unaffected by the type of protein used to supplement the culture medium. When embryos were thawed in medium containing NSA, both pregnancy (PR) and implantation rates (IR) were significantly lower (P <0.05) than if the medium was supplemented with serum (PR 8.3% and 17.5%; IR 4.6% and 10.5%). Inclusion of NSA before freezing reduced the IR of thawed embryos. To further test this observation all cycles where embryos were cultured and frozen in medium containing NSA (173 cycles) were matched to cycles where serum was used and the outcome was compared. At the end of 1995 just over half of the embryos in both groups had been thawed. No statistical difference was noted in the pregnancy rates (NSA, 5.6% versus serum, 11.3%) but the IR per embryo was significantly lower when embryos were cultured and frozen in medium supplemented with NSA (2.2%) than when serum was used as the supplement (6.6%).      

Elective single embryo transfer (eSET) policy in the first three IVF/ICSI treatment cycles

BACKGROUND: Elective single embryo transfer (eSET), applied in the first or second IVF cycle in young patients with good quality embryos, has been demonstrated to lower the twin pregnancy rate, while the overall pregnancy rate is not compromised. It is as yet unclear whether eSET could be the preferred transfer policy in all treatment cycles, or that it should be restricted to the first or first two cycles. METHODS: eSET policy (when two or more embryos were available, at least one of them being of good quality) was offered to patients younger than 38 years in the first three treatment cycles. Retrospectively, treatment cycle outcome was studied. RESULTS: In 326 patients, 586 treatment cycles were performed (326 first, 168 second and 92 third treatment cycles). In 65 cycles (11%), eSET could not be applied because there was either no fertilization, or only one embryo available. In the remaining 521 cycles, eSET was performed in 111 cycles (19%), while in 410 cycles, no good quality embryo was available resulting in the transfer of two embryos (double embryo transfer, DET). No significant differences in ongoing pregnancy rates after transfer of fresh embryos were observed between eSET and DET in the first (both 33%), second (36 and 23%, respectively) and third treatment cycles (20 and 24%, respectively). In significantly more eSET cycles compared to DET cycles, could embryos be frozen. This resulted in a significantly higher cumulative pregnancy rate after eSET compared to DET. CONCLUSIONS: In patients younger than 38 years with at least one top quality embryo, eSET can be the transfer policy of choice in at least the first three treatment cycles, since the pregnancy rates obtained in each treatment cycle are comparable to those after DET.     

超快速玻璃化冷冻人早期胚胎的临床应用价值初探

目的观察cryotop超快速玻璃化冷冻人早期胚胎的临床效果并探讨其应用价值。方法玻璃化冷冻试剂盒购自日本KITAZATO Biopharma公司,对新鲜周期胚胎移植后未妊娠患者的胚胎施行胚胎复苏,观察胚胎复苏后的存活率,胚胎分级和临床妊娠情况。结果共行胚胎复苏63个周期,201个胚胎,胚胎复苏后存活189个,胚胎复苏存活率为94.03%（189/201）;移植胚胎63个周期,移植胚胎总数187个,平均每周期移植胚胎2.97个（187/63）;临床妊娠30个周期,临床妊娠率为47.62%（30/63）,种植率为20.32%（38/187）,多胎率为23.33%（7/30）,流产率为6.67%（2/30）。结论 cryotop超快速玻璃化冷冻法简便且胚胎存活率高,是一种较好的冷冻人早期胚胎的方法。     

Cryopreservation of biopsied embryos at the blastocyst stage

BACKGROUND: The availability of an efficient cryopreservation program is especially important in the case of embryos that have undergone blastomere biopsy for PGD. Unfortunately, the freezing/thawing of biopsied embryos has given disappointing results when performed at the cleavage stage. In this study, embryos diagnosed as normal after PGD were grown to the blastocyst stage, frozen and thawed for successive frozen embryo transfer. METHODS: A total of 34 patients performed a thawing cycle in which 47 blastocysts were thawed. The cryopreservation solutions were based on HEPES-buffered medium supplemented with human serum albumin (HSA), sucrose and 1,2-propanediol. The same protocol was applied to embryos from 88 IVF/ICSI patients, which underwent 92 thawing cycles with 150 thawed blastocysts. RESULTS: The survival rate was similar in the two groups (53% after PGD and 58% in IVF/ICSI cycles), as well as the cumulative pregnancy rate per patient (59% after PGD versus 47% in IVF/ICSI cycles), despite a higher maternal age and a lower proportion of embryos available for transfer or cryopreservation in the PGD group. CONCLUSIONS: Neither the survival rate nor the subsequent development and chances of implantation, differed between embryos frozen at the blastocyst stage following biopsy and those frozen intact.     

Effect of developmental stage of embryo at freezing on pregnancy outcome of frozen-thawed embryo transfer

BACKGROUND: The study aim was to investigate the impact of the developmental stage of embryos on pregnancy outcome of frozen embryo transfer (FET). METHODS: The survival rates of embryos after thawing and pregnancy outcome following FET were compared retrospectively between three cryopreservation strategies utilizing either zygote, day 2 or day 3 embryo freezing. RESULTS: A total of 4006 embryos was analysed in 1657 thaw cycles. The highest (P < 0.0001) survival rate (all cells survived) was observed for zygotes (86.5%), followed by day 2 (61.7%) and day 3 (43.1%) embryos. FET was performed in 1586 (95.7%) of all thaw cycles, resulting in overall clinical pregnancy and implantation rates of 20.7 and 14.2% respectively. The delivery rate per transfer was 16.5%, and live birth rate per transferred embryo 11%. There were no significant differences in clinical pregnancy, implantation, delivery and birth rates between frozen zygote, day 2 and 3 embryo transfers. However, an elevated miscarriage rate was observed in the day 3 group (45%) compared with zygotes (21.3%; P = 0.049) and day 2 embryos (18.3%; P = 0.004). The overall efficacy of FET (birth rate per thawed embryo) was 7.3%. The efficacy was lower in day 3 group (4.2%) than in the zygote (7.1%; P = 0.082) and day 2 (7.6%; P = 0.027) groups. CONCLUSIONS: The developmental stage of embryos at freezing has a profound effect on their post-thaw survival, but seems to have little effect on rates of clinical pregnancy, implantation, delivery and birth after FET. The elevated miscarriage rate for day 3 frozen embryo transfers may be caused by damage during freeze-thaw procedures. The low survival rate and elevated miscarriage rate were both responsible for a reduced overall efficacy for day 3 FET when compared with zygotes and day 2 embryos.     

Clinical experience with ultrarapid cryopreservation of human embryos resulting from intracytoplasmic sperm injection: brief communication.

A total of 41 patients requested thawing of supernumerary embryos in an intracytoplasmic sperm injection (ICSI) programme. Mean patient age was 30.8 +/- 3.8 years. Embryo freezing by the ultrarapid method was performed at room temperature in 3 mol/l DMSO and 0.25 mol/l sucrose. Total freezing time was 2.5 min including filling of the straw. In the thawing process, the embryos were removed from liquid N(2), left at room temperature for 30 s, immersed for 40 s at 30 degrees C, and then successively transferred at room temperature for 10 min to each of three sucrose solutions of decreasing concentration. The embryos were kept in culture and only those that presented cleavage after 24 h were transferred. Embryos from 42 cycles were thawed and a total of 24 transfers was performed. The mean number of thawed embryos was 5.0 +/- 3.2 per cycle and the mean number of transferred embryos was 2.83 +/- 1.3. The clinical pregnancy rate per cycle obtained after the thawing process was 16. 6%. The clinical pregnancy rate per transfer was 29.2% and the implantation rate was 13.2%. The abortion rate was 14.3%. Six deliveries have been performed, with the birth of seven infants.