未來可能niPGT 與傳統TE biopy 可能同時並用以防止TE biopsy偵測失敗

Review

 

. 2025 Oct 14;14(20):1591.

 doi: 10.3390/cells14201591.

Noninvasive Preimplantation Genetic Testing in Recurrent Pregnancy Loss and Implantation Failure: Breakthrough or Overpromise?

Background: Recurrent pregnancy loss (RPL) and recurrent implantation failure (RIF) are significant challenges in reproductive medicine. For both, embryonic aneuploidy is the leading etiological factor. Preimplantation genetic testing for aneuploidy (PGT-A) via trophectoderm biopsy is the current standard for embryo selection. However, it is limited by its invasiveness, potential for embryo damage, and diagnostic errors due to mosaicism. Rationale/Objectives: This review critically evaluates the emerging role of noninvasive PGT (niPGT). NiPGT analyzes cell-free DNA from spent blastocyst culture media, thus, it is a potential alternative for managing RPL and RIF. Hence, the primary objective is to determine whether current evidence supports niPGT as a reliable replacement for conventional biopsy-based PGT-A in these high-risk populations. Outcomes: The analysis reveals that niPGT offers significant theoretical advantages. These include complete non-invasiveness, enhanced embryo preservation, and high patient acceptability. However, its clinical application is hampered by substantial limitations. Key amongst them is the inconsistent and often suboptimal diagnostic accuracy (sensitivity 70-85%, specificity 88-92%) compared to biopsy. Other significant factors include the high rates of amplification failure (10-50%), vulnerability to maternal DNA contamination, as well as low DNA yield. Crucially, there is a definitive lack of robust, prospective randomized controlled trial (RCT) data demonstrating improved live birth rates or reduced miscarriage rates specifically in RPL and RIF cohorts. As such, niPGT is not yet ready to be a standalone clinical adoption in RPL and RIF cases. However, it may serve as a valuable adjunct for rescue scenarios following biopsy failure or for ethical reasons

niPGT vs. 傳統切片PGT----對應全染色體，染色體一致率仍有差距 (77 vs 91%)

Ploidy concordance rates in SBM-TE, SBM-WB, and TE-WB were 77.4% (130/168), 77.4% (130/168), and 90.5% (152/168),

Multicenter Study

 

. 2025 Oct;42(10):3479-3491.

 doi: 10.1007/s10815-025-03670-8. Epub 2025 Oct 7.

Validation of non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) in a Japanese population: experience from seven clinics

Purpose: The aim of this study was to determine the feasibility of implementing non-invasive PGT-A (niPGT-A) in clinical practice. Informativity and concordance rates between three sample types (embryonic cell-free DNA [cfDNA] present in spent blastocyst medium [SBM], trophectoderm [TE] biopsy, and whole blastocyst [WB]) from the same embryo were evaluated.

Methods: This was a prospective, multicenter study conducted between February 2022 and November 2022 at seven Japanese IVF centres. 212 blastocysts were donated for research. The cfDNA released into the SBM was analysed, and the results were compared against the corresponding TE biopsy and WB sample.

Results: Overall informativity rates for SBM, TE, and WB were 81.6% (173/212), 98.6% (209/212), and 98.6% (209/212), respectively. There was no difference between TE and WB; however, SBM was significantly different to both (p < 0.001). The informativity rate in SBM samples significantly varied among the seven centres, ranging between 72.7 and 97.1% (p = 0.041). Ploidy concordance (SBM-TE) also varied across the centres, ranging between 68.2 and 90.9%; however, this did not reach statistical significance (p = 0.63). Ploidy concordance rates in SBM-TE, SBM-WB, and TE-WB were 77.4% (130/168), 77.4% (130/168), and 90.5% (152/168), respectively. There was no statistical difference for SBM-TE and SBM-WB, but both were statistically different to TE-WB (p = 0.0054).

Conclusion: We have shown results that are consistent with the existing literature, indicating the feasibility of applying the niPGT-A protocol described here, and thus using the SBM result to establish a priority for embryo transfer.

 ni-PGT培養模式(微滴10-15uL, D4-6不更換培養液) 

----對囊胚無明顯不良影響

Observational Study

 

. 2024 Sep 1;39(9):1952-1959.

 doi: 10.1093/humrep/deae156.

The impact of implementing a non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) embryo culture protocol on embryo viability and clinical outcomes

Study question: Are modifications in the embryo culture protocol needed to perform non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) affecting clinical reproductive outcomes, including blastocyst development and pregnancy outcomes?

Summary answer: The implementation of an embryo culture protocol to accommodate niPGT-A has no impact on blastocyst viability or pregnancy outcomes.

What is known already: The recent identification of embryo cell-free (cf) DNA in spent blastocyst media has created the possibility of simplifying PGT-A. Concerns, however, have arisen at two levels. First, the representativeness of that cfDNA to the real ploidy status of the embryo. Second, the logistical changes that need to be implemented by the IVF laboratory when performing niPGT-A and their effect on reproductive outcomes. Concordance rates of niPGT-A to invasive PGT-A have gradually improved; however, the impact of culture protocol changes is not as well understood.

Study design, size, duration: As part of a trial examining concordance rates of niPGT-A versus invasive PGT-A, the IVF clinics implemented a specific niPGT-A embryo culture protocol. Briefly, this involved initial culture of fertilized oocytes following each laboratory standard routine up to Day 4. On Day 4, embryos were washed and cultured individually in 10 μl of fresh media. On Day 6 or 7, blastocysts were then biopsied, vitrified, and media collected for the niPGT-A analysis. Six IVF clinics from the previously mentioned trial were enrolled in this analysis. In the concordance trial, Clinic A cultured all embryos (97 cycles and 355 embryos) up to Day 6 or 7, whereas in the remaining clinics (B-F) (379 cycles), nearly a quarter of all the blastocysts (231/985: 23.5%) were biopsied on Day 5, with the remaining blastocysts following the niPGT-A protocol (754/985: 76.5%). During the same period (April 2018-December 2020), the IVF clinics also performed standard invasive PGT-A, which involved culture of embryos up to Days 5, 6, or 7 when blastocysts were biopsied and vitrified.

Essure 使用於輸卵管水腫可能造成IVF胚胎植入流產

輸卵管切除仍應優先考慮於此類病患

 Essure 是一種曾被廣泛使用的「永久性避孕裝置」，由拜耳（Bayer）公司開發。它是一對細小的金屬彈簧線圈（由鎳鈦合金、不鏽鋼及聚對苯二甲酸乙二酯纖維製成），透過陰道植入輸卵管中，誘發組織發炎並產生疤痕（纖維化），進而永久阻塞輸卵管以達到避孕效

High miscarriage rate in women treated with Essure® for hydrosalpinx before embryo transfer: a systematic review and meta-analysis

Essure® has been tested as an alternative treatment for hydrosalpinx before embryo transfer (ET) in women undergoing assisted reproduction techniques. However, the persistence of a foreign body inside the uterine cavity might have a negative impact on the outcome of pregnancy. The present systematic review aimed at identifying, appraising and summarizing the available evidence regarding the effectiveness and safety of using Essure prior to ET for women with hydrosalpinx.

Methods

We searched for studies in PubMed, Scopus, CENTRAL, Web of Science and ClinicalTrials.gov and the reference lists of eligible studies. All studies including at least 10 women with hydrosalpinx who received Essure, any other intervention or no treatment prior to ET were considered eligible. Study selection, data extraction and evaluation of the risk of bias were performed independently by two authors. Study outcomes were miscarriage per clinical pregnancy, singleton preterm birth per singleton live birth and live birth/ongoing pregnancy and clinical pregnancy per ET. The pooled results for each outcome and intervention were summarized as proportions with their respective 95% CIs, using a random-effects model.

Results

Our electronic search of databases was performed on 7 November 2015, and 26 studies with 43 study arms were considered eligible: eight study arms evaluating Essure; seven assessing tubal aspiration; seven appraising effects of no treatment; 12 evaluating salpingectomy; two assessing tubal division; and seven evaluating tubal occlusion. When compared with women who had no intervention, women with Essure had a higher clinical pregnancy rate per ET (36% (95% CI, 0–43%) vs 13% (95% CI, 9–17%)). When compared with women who had other interventions, women with Essure had a higher miscarriage rate per clinical pregnancy (38% (95% CI, 27–49%) vs 15% (95% CI, 10–19%)).

Conclusions

The available evidence suggests that, although Essure prior to ET in women with hydrosalpinx improves the chance of achieving a clinical pregnancy compared with no intervention, it is associated with a higher rate of miscarriage when compared with the other interventions. Although this evidence is based on observational studies, we believe that salpingectomy should be the first option for women who are eligible for videolaparoscopy. However, it is still premature to make recommendations for women who are not eligible for surgery, and randomized controlled trials are needed to clarify which is the best treatment alternative in such a scenario. 

 試管嬰兒罹患先天心臟病機率高於自然懷孕嬰兒 (1.30% vs 0.68%)

ltrasound in Obstetrics & Gynecology

Congenital heart defects in IVF/ICSI pregnancy: systematic review and meta-analysis

There is no consensus in current practice guidelines on whether conception by in-vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) techniques is an indication for performing a fetal echocardiogram. The aim of the study was to assess whether congenital heart defects (CHD) occur more often in pregnancies conceived after IVF/ICSI as compared with those conceived spontaneously.

Methods

A systematic search for studies was conducted of PubMed/MEDLINE, EMBASE and Scopus from inception to September 2017. The search included the following medical subject heading (MeSH) terms alone or in different combinations: ‘IVF’, ‘IVF/ICSI’, ‘ART pregnancy’, ‘assisted conception’, ‘birth defect’, ‘congenital heart defects’ and ‘congenital malformation or abnormalities’. Studies comparing neonatal incidence of CHD in pregnancies conceived after IVF/ICSI and those conceived spontaneously were included. Studies reporting on other types of assisted reproductive technology (ART) or lacking information concerning termination of pregnancy were excluded. Chromosomal abnormalities were excluded in all analyzed studies. A meta-analysis of selected cohort studies was conducted to estimate the pooled odds ratio (OR) with 95% CI using a random-effects model. Statistical heterogeneity among the studies was evaluated with the I2 statistic and Q-test.

Results

Forty-one studies were identified for review including six case–control and 35 cohort studies. Data of eight selected cohort studies were used for meta-analysis. A total of 25 856 children conceived from IVF/ICSI techniques and 287 995 children conceived spontaneously, involving both singleton and multiple gestations, were included in the analysis. Total CHD events were 337/25 856 (1.30%) and 1952/287 995 (0.68%) in the IVF/ICSI and spontaneous conception groups, respectively. The risk of CHD was significantly increased in the IVF/ICSI group as compared with the spontaneous conception group (pooled OR, 1.45; 95% CI, 1.20–1.76; P = 0.0001; I2 = 44%; P = 0.08). In the subgroup of singleton IVF pregnancies, a significant difference was also obtained (OR, 1.55; 95% CI, 1.21–1.99; P = 0.0005; I2 = 36%; P = 0.18) and also multiple confounding factors adjusted ORs showed statistical significance (pooled OR, 1.29; 95% CI, 1.03–1.60; P = 0.02; I2 = 0%; P = 0.43).

Conclusion

Fetuses conceived with IVF/ICSI methods are at an increased risk of developing CHD compared with those conceived spontaneously. However, this finding deserves further investigation due to heterogeneity of both ART procedures and cardiac defects.  

在輔助生殖技術（ART）中，分開授精（Split Insemination） 主要指的是將取出的卵子分成兩組，分別使用不同的方法或精子來源進行受精。 

這種技術通常用於以下兩種主要情境：

1. 混合受精方式 (IVF + ICSI) 

這是最常見的臨床應用，又稱為「Half-ICSI」。實驗室會將同一批卵子（同胞卵子）隨機分成兩半： 

• 一組進行常規試管嬰兒 (IVF)： 讓精子與卵子在培養皿中自然結合。
• 另一組進行顯微注射 (ICSI)： 由胚胎師挑選單一精子直接注入卵子。 

主要目的：

• 防止受精失敗： 對於不明原因不孕或精子品質處於邊緣狀態（Borderline Semen）的夫婦，這能作為一種「保險」，確保至少有一組卵子能成功受精，避免整個週期無胚胎可用的窘境。
• 評估受精潛力： 觀察哪種方式效果更好，可作為未來治療週期的決策參考。
• 胚胎品質對比： 研究顯示，常規 IVF 產生的胚胎在某些情況下可能具有更好的囊胚發育潛能。 

•  與10% PVP組相比，5% PVP組精子DNA碎片化程度較低。
• 5% PVP組的受精率和高品質胚胎形成率也較高。
• 10% PVP組的碎片率、不均等卵裂球率、逆向卵裂（RC）率和胚胎停滯率均較高。
• ICSI週期中低濃度（5%）PVP可取代10% PVP。

The Beneficial Role of Low PVP Concentration on Sperm Apoptotic Gene Expression, Embryo Morphokinetics Status, and Clinical ICSI Outcomes

During the intracytoplasmic sperm injection (ICSI) procedure, 10% polyvinylpyrrolidone (PVP) is commonly used as a selection medium to decrease sperm motility. The study aimed to determine the effect of different concentrations of PVP (5% and 10%) on sperm apoptotic transcripts, sperm DNA fragmentation (SDF), embryo cytokinetic, and clinical characteristics in the ICSI program. In the study, 60 couples with male factor infertility underwent ICSI insemination using eitheconcentration. Metaphase II (MII) oocytes were divided into two groups, with one group injected with sperm in 5% PVP and the other in 10% PVP. After fertilization, the zygotes were cultured using a time-lapse microscope to assess morphokinetic parameters also, simultaneously with the ICSI procedure, SDF and apoptotic transcript levels were quantified using reverse transcription quantitative polymerase chain reaction (qPCR). The study found that DNA fragmentation was lower in sperm from the 5% PVP group compared to the 10% PVP group. Additionally, the expression levels of B-cell lymphoma (BCL2) and the 70 kilodalton heat shock proteins (HSP70) genes were significantly higher in 5% PVP compared to 10% PVP, while the transcript levels of the Bcl-2-associated X protein (BAX) gene were lower in 5% PVP. The 5% PVP group also demonstrated higher fertilization and high-quality embryo formation rates. Cytokinetic variables were significantly different between the two groups, and the rates of fragmentation, uneven blastomere, reverse cleavage (RC), and embryo arrest were higher in the 10% PVP group. The study concluded that a low concentration of PVP (5%) is a suitable replacement for 10% PVP in ICSI cycles for male factor infertility.

• 精子在PVP中暴露15、30和60分鐘後，其活力和形態均顯著受損，DNA碎片化和染色質結構異常增加，頂體反應精子的比例也隨之升高。此外，與未暴露於PVP的精子相比，粒線體膜電位高的精子數量顯著減少。
• 精子在PVP中暴露15分鐘後，PVP的有害作用顯著增強
• ICSI過程中，精子暴露於PVP的時間應限制在15分鐘內。

Prolonged exposure of human spermatozoa in polyvinylpyrrolidone has detrimental effects on sperm biological characteristics

Polyvinylpyrrolidone (PVP) has been utilized in intracytoplasmic sperm injection (ICSI) for immobilization and manipulation of spermatozoa. This study aims to determine the suitable time that sperm cells could be safely exposed to PVP during ICSI procedure. Twenty-five normal semen samples were prepared using the swim-up method and then were exposed to 10% PVP at different time intervals (15, 30 and 60 min). The effect of PVP on sperm parameters (viability and morphology), DNA fragmentation index (sperm chromatin dispersion test), chromatin quality (aniline blue, toluidine blue and chromomycin A3 staining), acrosome reaction, mitochondrial membrane potential and sperm ultrastructure was assessed at different time intervals. Our results showed that prolonged sperm exposure in PVP for 15, 30 and 60 min significantly affects viability and morphology with a concomitant increase in DNA fragmentation and abnormal chromatin structure, while the percentage of acrosome-reacted spermatozoa was additionally increased. In addition, the spermatozoa with high mitochondrial membrane potential were significantly decreased compared to unexposed spermatozoa to PVP. In conclusion, the detrimental effects of PVP were increased significantly following sperm exposure in PVP after 15 min. Therefore, the sperm exposure to PVP should be limited to less than 15 min during ICSI procedure.

 ICSI過程合併注射phospholipase Cζ (PLCζ)-cRNA 　可激活卵子提高受孕

可應用無精症精原細胞注射或體細胞注射複製胚胎

Birth of offspring from spermatid or somatic cell by co-injection of PLCζ-cRNA

Artificial oocyte activation is important for assisted reproductive technologies, such as fertilization with round spermatids (ROSI) or the production of cloned offspring by somatic cell nuclear transfer (SCNT). Recently, phospholipase Cζ (PLCζ)-cRNA was used to mimic the natural process of fertilization, but this method required the serial injection of PLCζ-cRNA and was found to cause damage to the manipulated oocytes. Here we tried to generate offspring derived from oocytes that were fertilized using round spermatid or somatic cell nuclear transfer with the co-injection of PLCζ-cRNA. After co-injecting round spermatids and 20 ng/µL of PLCζ-cRNA into the oocytes, most of them became activated, but the activation process was delayed by more than 1 h. With the co-injection method, the rate of blastocyst formation in ROSI embryos was higher (64%) compared with that of the serial injection method (55%). On another note, when SCNT was performed using the co-injection method, the cloned offspring were obtained with a higher success rate compared with the serial-injection method. However, in either ROSI or SCNT embryos, the birth rate of offspring via the co-injection method was similar to the Sr activation method. The epigenetic status of ROSI and SCNT zygotes that was examined showed no significant difference among all activation methods. The results indicated that although the PLCζ-cRNA co-injection method did not improve the production rate of offspring, this method simplified oocyte activation with less damage, and with accurate activation time in individual oocytes, it can be useful for the basic study of oocyte activation and development.

•  鋅 (Zn) 影響氧化壓力調節。
• 補充鋅可增加黃體體積，但降低血漿黃體素 (P4) 水平。
• 0.8 μg/ml Ｚn可降低孕酮合成和活性氧 (ROS) 水平，同時增強細胞活力；
• 而 1.2 μg/ml Ｚn無顯著影響。
• Zn以劑量依賴的方式調節黃體功能，降低氧化應激，同時抑制黃體素生成。
• Zn表現雙相效應，低濃度 (0.8 µg/ml) 可提供有益的抗氧化和保護作用（減少 ROS、促進孕酮/P4 合成、提高活力），
• 但稍高的劑量 (1.2 µg/ml) 會失去其積極作用，

Zinc (Zn) is an essential trace element for cellular processes such as oxidative stress regulation. 

In vivo, Zn supplementation increased CL size but reduced plasma P4 levels. 

In vitro, 0.8 μg/ml Zn decreased P4 synthesis and ROS levels while enhancing cell viability, whereas 1.2 μg/ml Zn had no significant effect compared to the control. 

Zn modulates luteal function in a dose-dependent manner, reducing oxidative stress while impairing P4 production.

 endometrial compaction: 植入胚胎錢４天　使用黃體素後，子宮內膜厚度略為下降＆緻密。

endometrial compaction 無法有效預測活產率

Review

 

. 2023 Nov;40(11):2513-2522.

 

Can endometrial compaction predict live birth rates in assisted reproductive technology cycles? A systematic review and meta-analysis

Purpose: Endometrial compaction (EC) is defined as the difference in endometrial thickness from the end of the follicular phase to the day of embryo transfer (ET). We aimed to determine the role of EC in predicting assisted reproductive technology (ART) success by conducting a meta-analysis of studies reporting the association between EC and clinical outcomes of ART.

Methods: MEDLINE via PubMed, Web of Science, Scopus, and the Cochrane Central Register of Controlled Trials (CENTRAL) were searched from the date of inception to May 19, 2023. The primary outcome was live birth rate (LBR) per ET. Secondary outcomes were live birth or ongoing pregnancy per ET, ongoing pregnancy per ET, clinical pregnancy per ET, and miscarriage per clinical pregnancy.

Results: Fifteen studies were included. When data from all studies reporting live birth were pooled, overall LBR rates were comparable in cycles showing EC or not [RR = 0.97, 95%CI = 0.92 to 1.02; 10 studies, 11,710 transfer cycles]. In a subgroup of studies that included euploid ET cycles, a similar LBR for patients with and without EC was noted [RR = 0.99, 95%CI = 0.86 to 1.13, 4 studies, 1172 cycles]. The miscarriage rate did not seem to be affected by the presence or absence of EC [RR = 1.06, 95%CI = 0.90 to 1.24; 12 studies].

Conclusion: The predictive value of EC in determining LBR is limited, and assessment of EC may no longer be necessary, given these findings.

 D７才長達囊胚著床率約５％

Review

 

. 2018 Jun 1;33(6):991-997.

 doi: 10.1093/humrep/dey091.

Should extended blastocyst culture include Day 7?

Extended culture to the blastocyst stage is widely practised, improving embryo selection and promoting single embryo transfer. Selection of useable blastocysts typically occurs on Days 5 and 6 of embryo culture. Embryos not suitable for transfer, biopsy or cryopreservation after Day 6 are routinely discarded. Some embryos develop at a slower rate, however, forming blastocysts on Day 7 of culture. Day 7 blastocysts can be viable, they can be of top morphological grade, euploid and result in a healthy live birth. Since ending culture on Day 6 is current practice in most clinics, viable Day 7 blastocysts may be prematurely discarded. Although Day 7 blastocysts make up only 5% of useable blastocysts, those which are suitable for cryopreservation or biopsy are clinically significant. Overall, culturing embryos an additional day increases the number of useable embryos per IVF cycle and provides further opportunity for pregnancy for patients, especially those who have only a few or low-quality blastocysts.

D7囊胚著床率只有４％

Ｄ5-6囊胚著床率有30+%

. 2024 Jul;41(7):1835-1842.

 doi: 10.1007/s10815-024-03129-2. Epub 2024 May 10.

Is a day 7 blastocyst predictive of the reproductive potential of sibling day 5 and day 6 blastocysts?

Methods: Retrospective cohort of autologous frozen embryo transfers (FET), February 2019 to April 2022. Cycles divided into groups 1 to 5, according to the day of embryo cryopreservation and the presence of a day 7 blastocyst sibling within the cohort: group 1/group 2-day 5 blastocyst without/with a day 7 sibling, group 3/group 4-day 6 blastocyst without/with a day 7 sibling, group 5-day 7 blastocyst. Clinical, ongoing pregnancy and miscarriage rates, cycle, and patient characteristics are reported. Multivariable generalized estimating equations (GEE) logistic regression analysis accounts for confounders and assesses the effect of a sibling day 7 blastocyst on ongoing pregnancy rates of day 5 or day 6 blastocyst FETs.

Results: Ongoing pregnancy rates are 38.4%, 59.5%, 30.8%, 32.7%, and 4.4% in groups 1-5, respectively. When correcting for maternal age, number of oocytes retrieved and discarded per cohort, and ploidy, embryos cryopreserved on either day 6 or day 7 have reduced odds of ongoing pregnancy after FET compared to day 5 blastocysts (OR = 0.76, IQR [0.61-0.95], p-value = 0.01). However, the presence of a day 7 sibling does not significantly affect odds of ongoing pregnancy of day 5 or day 6 blastocysts compared to the same-day blastocyst without a day 7 sibling (p-value = 0.20 and 0.46, respectively). This finding is consistent within both the Preimplantation Genetic Testing for Aneuploidy (PGT-A) unscreened and screened (euploid) embryo subgroups.

Conclusions: Day of embryo cryopreservation significantly affects ongoing pregnancy rates. However, day 7 embryos within a cohort do not affect the reproductive potential of sibling day 5 and day 6 blastocysts, suggesting that slow embryo development is an embryo-specific trait.