胚胎分裂速度遲滯  分裂非對稱 原因可能是染色體錯置異常

. 2019 Feb;36(2):315-324.

 doi: 10.1007/s10815-018-1361-8. Epub 2018 Nov 12.

Time-lapse imaging reveals delayed development of embryos carrying unbalanced chromosomal translocations

Purpose: The purpose of the study was to compare the morphokinetic parameters of embryos carrying balanced chromosomal translocations with those carrying unbalanced chromosomal translocations using time-lapse microscopy.

Methods: The study group included 270 embryos that underwent biopsies on day 3 for preimplantation genetic diagnosis (PGD) for chromosomal translocations in our unit between 2013 and 2015. All embryos were incubated under time-lapse microscopy and evaluated for timing of developmental events up to day 5. The timing of these events was compared between balanced and unbalanced embryos, potentially viable and nonviable variants, and maternal versus paternal inheritance of the translocation.

Results: The PGD analysis found that 209 (77%) of the 270 biopsied embryos carried an unbalanced translocation. Embryos carrying unbalanced translocations, which are expected to lead to implantation failure or miscarriage, cleaved less synchronously and were delayed in time of cleavage to the 4-cell stage (t4) and in time of start of blastulation (tSB) compared with balanced embryos (P < 0.05). Furthermore, embryos carrying nonviable translocations demonstrated a significant delay at the time of pronuclei fading (tPNf) compared with those carrying potentially viable translocations (P < 0.05). Embryos whose unbalanced translocations were of maternal origin were significantly delayed in most of the morphokinetic parameters (including tPNf, t2, t3, t4, t6, t7, t8, cc2, s2, and tSB) compared with embryos carrying balanced translocations (P < 0.05).

Conclusions: Embryos carrying unbalanced chromosomal translocations mainly of maternal origin undergo delayed development and asynchronous cleavage that may lead to implantation failure or miscarriage.

polar body 切片幾乎不會影響胚胎之發育&著床率

. 2018 Aug;35(8):1521-1528.

 doi: 10.1007/s10815-018-1207-4. Epub 2018 May 22.

Impact of polar body biopsy on embryo morphokinetics-back to the roots in preimplantation genetic testing?

Purpose: Polar body biopsy (PBB) is a common technique in preimplantation genetic testing (PGT) to assess the chromosomal status of the oocyte. Numerous studies have been implemented to investigate the impact of biopsies on embryo development; however, information on embryo morphokinetics is still lacking. Hence, we investigated the impact of PBB on morphokinetic parameters in early embryo development.

Methods: Four hundred four embryos (202 PBB, 202 control) were retrospectively analyzed. Patients were stimulated with a gonadotropin-releasing hormone antagonist ovarian hyperstimulation protocol. After fertilization check, embryos were incubated in a time-lapse incubator. The groups were matched for maternal age at time of oocyte retrieval.

Results: Mean group times for reaching specific developmental time points showed no significant difference comparing embryos with PBB conducted and without. Likewise, further subdivision of the PBB group in euploid and aneuploid embryos revealed no differences in the early embryo morphokinetic development compared to the control group. Aneuploidy testing revealed a high prevalence of chromosomal aberrations for chromosomes 21, 4, 16, and 19.

Conclusions: In conclusion, PBB does not impact the morphokinetic parameters of the embryo development. PBB can be safely applied without the risk of impairing the reproductive potential of the embryo and can be highly recommended as safe and practicable PGT approach, especially in countries with prevailing restrictions regarding PGT analysis.

胚胎切片在8 cell 晚期(post 8-cell 15-20h) 著床率優於8 cell 後早期(post 8 cell 0-15h)

. 2018 Jan 1;33(1):32-38.

 doi: 10.1093/humrep/dex343.

Optimal timing for blastomere biopsy of 8-cell embryos for preimplantation genetic diagnosis

Study question: What is the optimal timing for blastomere biopsy during the 8-cell stage, at which embryos will have the best implantation potential?

Summary answer: Fast-cleaving embryos that are biopsied during the last quarter (Q4) of the 8-cell stage and are less affected by the biopsy procedure, and their implantation potential is better than that of embryos biopsied earlier during the 8-cell stage (Q1-Q3).

What is known already: Blastomer biopsy from cleavage-stage embryos is usually performed on the morning of Day 3 when the embryos are at the 6- to 8-cell stage and is still the preferred biopsy method for preimplantation genetic diagnosis (PGD) for monogentic disorders or chromosomal translocations. Human embryos usually remain at the 8-cell stage for a relatively long 'arrest phase' in which cells grow, duplicate their DNA and synthesize various proteins in preparation for the subsequent division.

Study design, size, duration: This is a retrospective cohort study. The study group (195 embryos) included all 8-cell stage embryos that underwent blastomere biopsy for PGD for monogenetic disorders and chromosomal translocations in our unit between 2012-2014 and cultured in the EmbryoScope until transfer. The control group (115 embryos) included all embryos that underwent intracytoplasmic sperm injection without a biopsy during the same period.

Participants/materials, setting, methods: The 8-cell stage was divided into four quarters: the first 5 h post-t8 (Q1), 5-10 h post-t8 (Q2), 10-15 h post-t8 (Q3) and at 15-20 h post-t8 (Q4). Non-biopsied control embryos were divided into four equivalent quarters. Embryos were evaluated for timing of developmental events following biopsy including timing of first cleavge after biopsy, timing of comapction (tM) and start of blastulation (tSB). Timing of these events were compared between PGD and control embryos, as well as with 56 PGD implanted embryos with Known Implantation Data (PGD-KID-positive embryos).

Main results and the role of chance: Embryos that were biopsied during Q3 (10-15 h from entry into 8-cell stage) were delayed in all three subsequent developmental events, including first cleavage after biopsy, compaction and start of blastulation. In contrast, these events occurred exactly at the same time as in the control group, in embryos that were biopsied during Q1, Q2 or Q4 of the 8-cell stage. The results show also that embryos that were biopsied during Q1, Q2 or Q3 of the 8-cell stage demonstrated a significant delay from the biopsied implanted embryos already in t8 as well as in tM and tSB. However, embryos that were biopsied during Q4 demonstrated dynamics similar to those of the biopsied implanted embryos in t8 and tM, and a delay was noticed only in the last stage of tSB.

PRP(platelet‐rich plasma)  可能可以提高反復流產病患之子宮內膜之厚度

• Intrauterine administration of platelet‐rich plasma improves embryo implantation by increasing the endometrial thickness in women with repeated implantation failure: A single‐arm self‐controlled trial

  • Maki Kusumi
   
  • Tatsuji Ihana
   
  • Takako Kurosawa
   
  • Yasuo Ohashi
   
  • Osamu Tsutsumi

  Reproductive Medicine and BiologyEarly View

  First published: 25 June 2020

  Abstract

  Abstract

  Purpose

  The purpose of this study was to investigate the effectiveness of intrauterine administration of platelet‐rich plasma (PRP) in frozen embryo transfer (FET) cycle in Japanese patients with a thin endometrium.

  Method

  A prospective single‐arm self‐controlled trial was conducted in Japan. PRP administration was performed in 36 of the 39 eligible patients with a thin endometrium (≤7 mm). Hormone replacement therapy (HRT) with estrogen was performed for 2 menstrual cycles, and PRP was administrated on the 10th and 12th days of the second HRT cycle. The endometrial thickness was evaluated on transvaginal ultrasonography by two physicians at every visit, one an attending physician and the other a specialist physician blinded to the date and timing of the sonography. FET was performed during the second HRT cycle after PRP administration.

  Results

  After PRP administration, the mean (SD) endometrial thickness on the 14th day was significantly increased by 1.27 mm (P  < .001) and 0.72 mm (P  = .001) on the basis of the unblinded and blinded measurements, respectively. Of the 36 patients, 32 (88.9%) underwent FET. The clinical pregnancy rate was 15.6%. No adverse events occurred.

  Conclusions

  PRP therapy was safe and effective in increasing endometrial thickness improving possibly pregnancy rate.

睪丸萎縮病患施行TESE失敗率較高  較容易取不到精蟲

Testicular volume in non‐obstructive azoospermia with a history of bilateral cryptorchidism may predict successful sperm retrieval by testicular sperm extraction

Akiyoshi Osaka 

 

Toshiyuki Iwahata 

 

Yoshitomo Kobori 

 

Yukihito Shimomura 

 

Naoki Yoshikawa 

 

Shin Onota 

 

Atsushi Yamamoto 

 

Hisamitsu Ide 

 

Kouhei Sugimoto 

 

Hiroshi Okada

First published: 12 July 2020

 

https://doi.org/10.1002/rmb2.12338

Purpose

Cryptorchidism is one of the most common causes of non‐obstructive azoospermia (NOA) in adulthood. Even if early orchidopexy is performed to preserve fertility potential, some patients still suffer from azoospermia. Fertility potential is significantly lower in bilateral than unilateral cryptorchidism. The aims of this study were to identify clinical parameters that predict the likely success of sperm recovery by microscopic testicular sperm extraction (micro‐TESE) and also the likely outcome of intracytoplasmic sperm injection using sperm from NOA patients who submitted to bilateral orchidopexy.

Methods

Fifty‐two NOA patients with a history of bilateral cryptorchidism underwent micro‐TESE. The following clinical parameters were evaluated as predictive factors for successful sperm recovery: age at micro‐TESE; age at orchidopexy; period from orchidopexy to micro‐TESE; luteinizing hormone (LH); follicle‐stimulating hormone (FSH); testosterone; average testicular volume; and body mass index.

Results

In the successful sperm retrieval group, average testicular volume was significantly greater, while serum LH and FSH, and body mass index were significantly lower. In a multivariate analysis, average testicular volume was positively correlated with successful sperm recovery.

Conclusion

Our results indicate that testicular volume in NOA patients with bilateral cryptorchidism is a predictor for successful sperm recovery.

cryotop玻璃化冷凍    明顯優於    毛細管玻璃化冷凍hollow fiber vitrification

. 2019 Dec 21;19(2):142-150.

 doi: 10.1002/rmb2.12312. eCollection 2020 Apr.

Hollow fiber vitrification allows cryopreservation of embryos with compromised cryotolerance

Purpose: This study aims to demonstrate vitrification methods that provide reliable cryopreservation for embryos with compromised cryotolerance.

Methods: Two-cell stage mouse embryos and in vitro produced porcine embryos were vitrified using the hollow fiber vitrification (HFV) and Cryotop (CT) methods. The performance of these two methods was compared by the viability of the vitrified-rewarmed embryos.

Results: Regardless of the method used, 100% of the mouse 2-cell embryos developed successfully after vitrification-rewarming into the blastocyst stage, whereas vitrification tests using porcine morulae with the HFV method produced significantly better results. The developmental rates of vitrified porcine morula into the blastocyst stage, as well as blastocyst cell number, were 90.3% and 112.3 ± 6.9 in the HFV group compared with 63.4% and 89.5 ± 8.1 in the CT group (P < .05). Vitrification tests using 4- to 8-cell porcine embryos resulted in development into the blastocyst stage (45.5%) in the HFV group alone, demonstrating its better efficacy. The HFV method did not impair embryo viability, even after spontaneous rewarming at room temperature for vitrified embryos, which is generally considered a contraindication.

Conclusion: Vitrification test using embryos with compromised cryotolerance allows for more precise determining of effective cryopreservation methods and devices.

D6囊胚植入懷孕率明顯低於D5囊胚植入

. 2019 Oct 2;34(10):1948-1964.

 doi: 10.1093/humrep/dez163.

Day 5 versus Day 6 blastocyst transfers: a systematic review and meta-analysis of clinical outcomes

Study question: Is there a difference in clinical pregnancy and live birth rates (LBRs) between blastocysts developing on Day 5 (D5) and blastocysts developing on Day 6 (D6) following fresh and frozen transfers?

Summary answer: D5 blastocyst transfers (BTs) present higher clinical pregnancy and LBRs than D6 in both fresh and frozen transfers.

What is known already: BT is increasingly popular in assisted reproductive technology (ART) centers today. To our knowledge, no meta-analysis has focused on clinical outcomes in both fresh and frozen BT. Concerning frozen blastocysts, one meta-analysis in 2010 found no significant difference in pregnancy outcomes between D5 and D6 BT. Since then, ART practices have evolved particularly with the wide use of vitrification, and more articles comparing D5 and D6 BT cycles have been published and described conflicting results.

Study design, size, duration: Systematic review and meta-analysis of published controlled studies. Searches were conducted from 2005 to February 2018 on MEDLINE and Cochrane Library and from 2005 to May 2017 on EMBASE, Eudract and clinicaltrials.gov, using the following search terms: blastocyst, Day 5, Day 6, pregnancy, implantation, live birth and embryo transfer (ET).

Participants/materials, setting, methods: A total of 47 full-text articles were preselected from 808 references, based on title and abstract and assessed utilizing the Newcastle-Ottowa Quality Assessment Scales. Study selection and data extraction were carried out by two independent reviewers according to Cochrane methods. Random-effect meta-analysis was performed on all data (overall analysis) followed by subgroup analysis (fresh, vitrified/warmed, slow frozen/thawed).

Main results and the role of chance: Data from 29 relevant articles were extracted and integrated in the meta-analysis. Meta-analysis of the 23 studies that reported clinical pregnancy rate (CPR) as an outcome, including overall fresh and/or frozen ET cycles, showed a significantly higher CPR following D5 ET compared with D6 ET (risk ratio (RR) = 1.27, 95% CI: 1.15-1.39, P < 0.001). For CPR, calculated subgroup RRs were 2.38 (95% CI: 1.74-3.24, P < 0.001) for fresh BT; 1.27 (95% CI: 1.16-1.39, P < 0.001) for vitrified/warmed BT; and 1.15 (95% CI: 0.93-1.41, P = 0.20) for slow frozen/thawed BT. LBR was also significantly higher after D5 BT (overall RR = 1.50 (95% CI: 1.32-1.69), P < 0.001). The LBR calculated RRs for subgroups were 1.74 (95% CI: 1.37-2.20, P < 0.001) for fresh BT; 1.38 (95% CI: 1.23-1.56, P < 0.001) for vitrified/warmed BT; and 1.44 (95% CI: 0.70-2.96, P = 0.32) for slow frozen/thawed BT. Sensitivity analysis led to similar results and conclusions: CPR and LBR were significantly higher following D5 compared to D6 BT.

大規模統計
全冷凍(freeze-all)解凍胚胎植入並無明顯較高懷孕率

Review

 

. 2020 Feb 1;92:9-14.

 doi: 10.1016/j.cryobiol.2019.11.041. Epub 2019 Nov 23.

Clinical utility of freeze-all approach in ART treatment: A mini-review

Romualdo Sciorio 1, Sandro C Esteves 2

A significant proportion of couples at reproductive age rely on assisted reproductive technology to overcome infertility. In vitro fertilisation (IVF) involves typically the use of exogenous gonadotropins to stimulate the ovary to produce oocytes, which are collected surgically. After fertilization by conventional IVF or intracytoplasmic sperm injection (ICSI), embryos are cultured in the embryology laboratory for a few days before being replaced into the uterus (fresh embryo transfer). Spare embryos can be vitrified and stored in liquid nitrogen to be transferred in a subsequent cycle. Over the years, concerns have arisen about possible adverse outcomes of transferring embryos back to the uterus immediately after controlled ovarian stimulation (COS) as regards to obstetrical and perinatal outcomes. It has been suggested that high hormonal levels during COS could create a relatively hostile environment for embryo implantation whilst increasing the risk of ovarian hyperstimulation syndrome (OHSS). With the remarkable improvement of vitrification as an alternative to the slow-freezing technique for human embryos, a new strategy the so-called "freeze-all" (FA) or "elective frozen embryo transfer" (eFET) was introduced. This approach involves COS, followed by the elective cryopreservation of the entire cohort of viable embryos to be transferred to the uterus in subsequent cycles in a possibly more physiological environment, thus avoiding the supra-physiologic hormonal levels observed during COS. The initial reports suggested that this policy could lead to improved pregnancy rates and reduced perinatal complications, which resulted in a steady increase and widespread use of FA globally. However, as data accumulated, it became clear that the use of FA to unselected couples undergoing ART offered no additional benefits over the conventional approach. Nonetheless, current evidence based on randomized controlled trials and observational studies indicates that FA might be justified in selected clinical scenarios, such as those involving the risk of OHSS. By contrast, there is a lack of evidence to support the FA policy for other indications, such as implantation failure or high progesterone levels on the trigger day. This review summarizes the clinical effectiveness of FA with the main focus on the health of offspring.

PGS 懷孕病患之HCG 平均較未作embryo biopsy之HCG為低

Trophectoderm biopsy—perhaps not such a benign intervention

Catherine E. Gordon, M.D.

, 

Catherine Racowsky, Ph.D.

Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women’s Hospital, Harvard Medical School, Boston, Massachusetts

DOI: https://doi.org/10.1016/j.fertnstert.2020.06.027

Article Info

• Abstract

• Full Text

• References

In this issue of Fertility and Sterility, Lu et al. (1) conducted a retrospective cohort study to investigate whether the removal of a few trophectoderm cells during biopsy for preimplantation genetic testing (PGT) was associated with a decrease in serum human chorionic gonadotropin (β-hCG) level 12 days after transfer and/or any adverse perinatal outcomes. Comparison of clinical pregnancies after cryoembryo transfer of biopsied versus nonbiopsied single embryos showed that the biopsy group had statistically significantly lower mean β-hCG levels (703.1 ± 569.6 vs. 809.2 ± 582.0 mIU/mL; P=.004). The authors also noted a lower threshold serum β-hCG level in the biopsy group as compared with the control group for prediction of live birth (368.6 mIU/mL [area under the curve 0.79] vs. 411.5 mIU/mL for the controls). There was no difference in miscarriage rates, live-birth rates, or perinatal outcomes between the two groups.

We applaud the authors for this contribution to the literature. With the ever-increasing use of trophectoderm biopsy for preimplantation genetic testing for aneuploidy (PGT-A), it is important to understand how this invasive manipulation might affect pregnancy outcomes. The questions the authors pose are logical in that we know that β-hCG is secreted by differential syncytiotrophoblasts at the site of implantation, and serum β-hCG levels after embryo transfer are correlated with pregnancy outcome. Although previous studies have shown that implantation and pregnancy rates are similar between biopsied and nonbiopsied embryos (2), the data remain sparse regarding any association between this technology and perinatal outcomes. Any adverse long-term associations to the children born from trophectoderm biopsied embryos are currently unknown.

Many studies have attempted to determine the optimal initial β-hCG levels and threshold values for prediction of successful outcomes after embryo transfer. However, the day of β-hCG measurement and type of assay used have varied greatly between studies. A strength of the study by Lu et al. (1) is that although the cycles were performed over a 2-year period, the same assay platform was used throughout, and all β-hCG measurements were taken on day 12 after embryo transfer. Together, these methodologies allowed an unbiased comparison of the β-hCG values to be made between the unbiopsied and biopsied groups.

Previous investigations have shown that a single β-hCG value for predicting pregnancy outcome may have less value than serial measurements. Therefore, it would have been informative if Lu et al. had compared the trends in β-hCG rise for each of the two groups, derived from at least two measurements. Additionally, despite the observed statistically significant differences in the mean β-hCG values for each group, the standard deviations were large and significantly overlapped, which raises questions regarding the relevance of the authors’ findings to the clinical management of the patient. A much reduced β-hCG value in the biopsy group compared with the control group, combined with similar pregnancy outcomes, would have provided greater assurance to providers that pregnancies with low β-hCG levels after trophectoderm biopsy are still capable of progressing to a live birth. Although the study included more than 800 pregnancies, a larger data set is required to determine whether a lower β-hCG cutoff value predicts a successful pregnancy after trophectoderm biopsy.

We note that there were more programmed embryo transfer cycles in the biopsy group as compared with the control group. Although the authors controlled for this (as well as for other variables) in their generalized linear regression analyses, it remains to be seen whether increased use of a programmed endometrial preparation in the biopsy group had any impact on β-hCG levels.

PGT無法提高IVF整體懷孕率

Randomized Controlled Trial

 

. 2019 Dec;112(6):1071-1079.e7.

 doi: 10.1016/j.fertnstert.2019.07.1346. Epub 2019 Sep 21.

Preimplantation genetic testing for aneuploidy versus morphology as selection criteria for single frozen-thawed embryo transfer in good-prognosis patients: a multicenter randomized clinical trial

Objective: To evaluate the benefit of next-generation sequencing (NGS)-based preimplantation genetic testing for aneuploidy (PGT-A) for embryo selection in frozen-thawed embryo transfer.

Patient(s): Women aged 25-40 years undergoing IVF with at least two blastocysts that could be biopsied.

Intervention(s): Randomization for single frozen-thawed embryo transfer with embryo selection based on PGT-A euploid status versus morphology.

Main outcome measure(s): Ongoing pregnancy rate (OPR) at 20 weeks' gestation per embryo transfer.

Result(s): A total of 661 women (average age 33.7 ± 3.6 years) were randomized to PGT-A (n = 330) or morphology alone (n = 331). The OPR was equivalent between the two arms, with no significant difference per embryo transfer (50% [137/274] vs. 46% [143/313]) or per intention to treat (ITT) at randomization (41.8% [138/330] vs. 43.5% [144/331]). Post hoc analysis of women aged 35-40 years showed a significant increase in OPR per embryo transfer (51% [62/122] vs. 37% [54/145]) but not per ITT.

Conclusion(s): PGT-A did not improve overall pregnancy outcomes in all women, as analyzed per embryo transfer or per ITT. There was a significant increase in OPR per embryo transfer with the use of PGT-A in the subgroup of women aged 35-40 years who had two or more embryos that could be biopsied, but this was not significant when analyzed by ITT.

PGS檢測異常之囊胚植入後仍有近一半49%胚胎著床活產  流產率只有9%

. 2019 Aug;36(8):1599-1607.

 doi: 10.1007/s10815-019-01510-0. Epub 2019 Jun 24.

Worldwide live births following the transfer of chromosomally "Abnormal" embryos after PGT/A: results of a worldwide web-based survey

Pasquale Patrizio 1, Gon Shoham 2, Zeev Shoham 3 4, Milton Leong 5, David H Barad 6 7, Norbert Gleicher 6 7 8 9

Purpose: Preimplantation genetic testing for aneuploidy (PGT-A) has become increasingly controversial since normal euploid births have been reported following transfer of embryos diagnosed as "abnormal." There is an increasing trend in transferring "abnormal" embryos; but it is still unknown how many IVF centers transfer "abnormal" embryos and with what efficiency.

Methods: We performed a worldwide web-survey of IVF centers to elucidate PGT-A related practice patterns including transfer of human embryos found "abnormal" by PGT-A. Participating centers reflected in vitro fertilization (IVF) cycles in the USA, Canada, Europe, Asia, South America, and Africa.

Results: One hundred fifty-one IVF centers completed the survey; 125 (83%) reported utilization of PGT-A. Europe had the highest utilization (32.3%), followed by the USA and Canada combined at 29.1%. The leading indications for PGT-A were advanced maternal age (77%), followed by recurrent implantation failure (70%), unexplained pregnancy loss (65%), and sex determination (25%); 14% of respondents used PGT-A for all of their IVF cycles; 20% of IVF units reported transfers of chromosomally "abnormal" embryos, and 56% of these took place in the USA, followed by Asia in 20%. Remarkably, 106 (49.3%) cycles resulted in ongoing pregnancies (n = 50) or live births (n = 56). Miscarriages were rare (n = 20; 9.3%).

Conclusions: The transfers of "abnormal" embryos by PGT-A offered robust pregnancy and live birth chances with low miscarriage rates. These data further strengthen the argument that PGT-A cannot reliably determine which embryos should or should not be transferred and leads to disposal of many normal embryos with excellent pregnancy potential.