傳統培養箱  vs  time lapse培養

懷孕率無差異

. 2018 Feb;109(2):302-309.e1.

 doi: 10.1016/j.fertnstert.2017.10.008. Epub 2017 Nov 23.

Randomized controlled trial comparing embryo culture in two incubator systems: G185 K-System versus EmbryoScope

Objective: To study whether the closed culture system, as compared with a benchtop incubator with similar culture conditions, has a positive impact on intracytoplasmic sperm injection (ICSI) outcomes.

Patient(s): A total of 386 patients undergoing ICSI cycles with at least six mature oocytes were randomized.

Intervention(s): Of these patients, 195 were assigned to the group with culture in a time-lapse imaging (TLI) system (EmbryoScope) and 191 to the group with culture in the G185 K-System (G185).

Main outcome measure(s): Rate of implantation (primary endpoint) and embryo morphology grade.

Result(s): No significant differences were found in the implantation rates. The proportion of high-grade embryos on day 2 was significantly higher in the TLI group compared with the G185 group (40.4% vs. 35.2%). The impact of the incubator on embryo morphology remained significant in multivariate analysis, which took into account the woman's age, the rank of attempt, and the smoking status (TLI vs. G185: odds ratio = 1.27; 95% confidence interval, [1.04-1.55]). No difference was found in the mean number of frozen embryos, even though the total proportion of frozen embryos was significantly higher in the TLI group than in the G185 group (29.5% vs. 24.8%).

Conclusion(s): No difference in implantation rate was found between the two incubators for fresh cycles. It remains to be determined whether the observed differences in embryo morphology and the total number of embryos cryopreserved would translate into higher cumulative outcomes with subsequent frozen embryo transfers.

 time-lapse(不取出觀察) vs傳統培養箱(定期取出觀察)

胚胎DNA甲基化程度並無差異(DNA甲基化程度越高代表外界干擾程度越大)

結論: 

胚胎有一定抵抗外界環境變動之能力  短暫取出觀察溫度環境變動對胚胎影響並不太大

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. 2023 Jan;40(1):113-123.

 doi: 10.1007/s10815-022-02669-9. Epub 2022 Dec 2.

Comparison of DNA methylation profiles of human embryos cultured in either uninterrupted or interrupted incubators

Purpose: We aimed to compare the DNA methylation profiles of human embryos cultured in uninterrupted or interrupted incubators.

Methods: This study included 9 women, ≤ 30 years old (range: 20-30 years), without a history of genetic diseases or smoking, undergoing ICSI treatment, and each woman donated one oocyte. Embryos were randomly assigned to culture in either time-lapse imaging or standard incubators after ICSI. We compared the DNA methylation profiles of human eight-cell embryos cultured in uninterrupted condition using time-lapse imaging (TLI) incubator (EmbryoScope) to those cultured in interrupted culture model using standard incubators (SI, G185 K-System). Nine single-cell whole-genome bisulfite sequencing (WGBS) datasets were analyzed, including four SI-cultured embryos and five TLI-cultured embryos at the eight-cell stage.

Results: A total of 581,140,020 and 732,348,182 clean reads were generated from the TLI and SI groups, respectively. TLI-cultured embryos had similar genome-wide methylation patterns to SI-cultured embryos. There were no significant differences in the methylation and transcription levels of transposable elements and imprinted control regions. Although a total of 198 differentially methylated genes (DMGs) were identified, only five DMGs had significantly different transcription levels between the two groups.

Conclusions: This is the first study to compare the DNA methylation profiles of embryos cultured in TLI and SI and will provide a foundation for evaluating the safety of TLI application in assisted reproductive technologies. However, further study with a larger cohort of samples was needed for the data validation.

PGS是否提高懷孕率仍具爭議  

本篇指出年輕ㄉIVF施行PGS並無明顯提升累積懷孕率(甚至有下降疑慮)

. 2023 Jan;40(1):137-149.

 doi: 10.1007/s10815-022-02667-x. Epub 2022 Dec 1.

PGT-A is associated with reduced cumulative live birth rate in first reported IVF stimulation cycles age ≤ 40: an analysis of 133,494 autologous cycles reported to SART CORS

Purpose: To evaluate the impact of preimplantation genetic testing for aneuploidy (PGT-A) on cumulative live birth rate (CLBR) in IVF cycles.

Methods: Retrospective cohort study of the SART CORS database, comparing CLBR for patients using autologous oocytes, with or without PGT-A. The first reported autologous ovarian stimulation cycle per patient between January 1, 2014, and December 31, 2015, and all linked embryo transfer cycles between January 1, 2014, and December 31, 2016, were included in the study. Exclusion criteria were donor oocyte cycles, donor embryo cycles, gestational carrier cycles, cycles which included both a fresh embryo transfer (ET) combined with a thawed embryo previously frozen (ET plus FET), or cycles with a fresh ET after PGT-A.

Results: A total of 133,494 autologous IVF cycles were analyzed. Amongst patients who had blastocysts available for either ET or PGT-A, including those without transferrable embryos, decreased CLBR was noted in the PGT-A group at all ages, except ages > 40 (p < 0.01). A subgroup analysis of only those patients who had PGT-A and a subsequent FET, excluding those without transferrable embryos, demonstrated a very high CLBR, ranging from 71.2% at age < 35 to 50.2% at age > 42. Rates of multiple gestations, preterm birth, early pregnancy loss, and low birth weight were all greater in the non-PGT-A group.

Conclusions: PGT-A was associated with decreased CLBR amongst all patients who had blastocysts available for ET or PGT-A, except those aged > 40. The negative association of PGT-A use and CLBR per cycle start was especially pronounced at age < 35.

 取卵過程汙染造成卵泡液微生物會導致懷孕率下降  (19.7% vs. 32.2%)

Review

 

. 2023 Nov;40(11):2501-2511.

 doi: 10.1007/s10815-023-02912-x. Epub 2023 Sep 9.

Associations between microbial presence in follicular fluid with IVF outcomes: a systematic review and meta-analysis

Purpose: The aim of the study was to synthesize disparate studies to investigate potential impact of microbial presence in FF of infertile women on IVF outcomes.

Methods: Following preliminary searches to find medical subject heading (MeSH) terms plus free terms, a systematic search was performed in the PubMed, Cochrane Library, Embase, Web of Science, and Clinicaltrials.gov databases from January 10, 2022, to July 5, 2023. Data collected for each study were analyzed using RevMan 5.4 software available on the Cochrane website.

Results: After correcting for contamination from the vagina, the FFs of 289 women were detected positively by microbial culture and identification, ELISA, and IPA. The pregnancy rate of the FF-positive group was significantly lower than the FF-negative group (19.7% vs. 32.2%) and (OR: 0.57, 95% CI: 0.28-1.14, P=0.11; I2=56%) while the fertilization rate was almost equal (60.0% vs. 62.0%) and (OR: 1.03, 95% CI: 0.88-1.20, P=0.72; I2=0%). Evidence quality was very low.

Conclusions: The different species of microorganisms in FF of infertile women may have different effects on IVF outcomes. The Lactobacillus spp. may have a positive effect, while other microorganisms may have the opposite effect.

 Letrozole 使用於冷凍胚胎植入療程可能可下降胎兒生長過重& 懷孕高血壓

. 2023 Dec;40(12):2885-2894.

 doi: 10.1007/s10815-023-02956-z. Epub 2023 Oct 10.

Letrozole use in vitrified single-blastocyst transfer cycles is associated with lower risk of large for gestational age infants in patients with polycystic ovary syndrome

Purpose: To evaluate the obstetric and perinatal outcomes of three routine endometrial preparation protocols in women with PCOS who underwent frozen embryo transfer (FET).

Methods: This was a retrospective study in women with PCOS who underwent FET in an academic reproductive medical center. A total of 2710 cycles were enrolled and classified into three groups according to different endometrial preparation protocols; human menopausal gonadotropin (HMG), letrozole + HMG, or hormone replacement therapy (HRT).

Results: The stimulation groups had reduced risks of hypertensive disorders of pregnancy (HDP), large for gestational age (LGA) infants, and cesarean delivery than the HRT group. After adjustment for different confounder combinations in the two models, the frequencies of LGA and HDP in the letrozole + HMG group and the HMG group were still significantly lower than those in the HRT group. The letrozole + HMG group exhibited a reduced risk of LGA than HMG group after adjustment of confounders. A trend toward risk reductions in HDP and LGA was observe in turns of HRT, HMG, and letrozole + HMG groups, and the trends were statistically significant (Ptrend = 0.031 and 0.001).

Conclusion: In patients with PCOS, ovarian stimulation protocols for endometrial preparation are associated with reduced risks of HDP and LGA compared to HRT cycles. The use of letrozole could further reduce risk of LGA compared to HMG only protocol. We propose that ovarian stimulation protocols can be used widely for endometrial preparation in FET cycles in women with PCOS, especially with the use of letrozole.

 2段式胚胎植入( afterload catheters 先放外管後 再吸取胚胎植入內管)  

懷孕率優於

1段式胚胎植入(direct catheters 外管內管同時一起植入)

. 2023 Dec;40(12):2895-2902.

 doi: 10.1007/s10815-023-02957-y. Epub 2023 Oct 11.

Steps forward in embryo transfer technique: a retrospective study comparing direct versus afterload catheters at different time frames

Purpose: To assess whether embryo transfer (ET) technique can influence the clinical pregnancy rate (CPR) and its correlation with the embryo transfer difficulty.

Design: This single center retrospective cohort analysis of fresh and frozen single blastocyst transfers performed between January 2016 and December 2021 included fresh and frozen single blastocyst transfers performed during the study timeframe. Direct technique was the only one used from January 2016 to September 2017. From September 2017 to March 2019, the choice between the two techniques was given by randomization, due to a clinical trial recruitment. From April 2019, only the afterload technique was used. Preimplantation genetic testing cycles and gamete donation procedures and cycles performed with external gametes or embryos were excluded. CPR was the primary outcome, while difficult transfer rate the secondary one. Univariate and multivariate logistic regressions were performed.

Results: During the period, 8,189 transfers were performed. CPR of the afterload group resulted significantly higher compared to the direct group (44.69% versus 41.65%, OR 1.13, 95% CI 1.02-1.25, p = 0.017) and the rate of difficult transfers two-thirds lower (9.06% versus 26.85%, OR 0.27, 95% CI 0.24-0.31, p < 0.001).

Conclusion: Our study demonstrated that CPR is significantly affected by the ET technique. In particular, with the afterload protocol, both CPR and easy transfer rates increased.

雷射Laser輔助孵化可輕微提高卵裂期胚胎凍融胚胎移植週期的懷孕率

Laser vs Non-Laser 

臨床懷孕率40.7% vs. 38.3%

活產率34.9% vs. 31.4% 

Laser-assisted hatching improves pregnancy outcomes in frozen-thawed embryo transfer cycles of cleavage-stage embryos

J Assist Reprod Genet. 2023 Feb; 40(2): 417–427.

Published online 2023 Jan 7. doi: 10.1007/s10815-022-02711-w

PMCID: PMC9935798

PMID: 36609944

Laser-assisted hatching improves pregnancy outcomes in frozen-thawed embryo transfer cycles of cleavage-stage embryos: a large retrospective cohort study with propensity score matching

Introduction

Laser-assisted hatching (LAH) is a commonly used adjunct technique; however, its effectiveness has not been fully established.

Objective

We evaluated the effects of LAH on pregnancy outcomes in frozen-thawed embryo transfer (FET) cycles of cleavage-stage embryos.

Materials and methods

This retrospective study involved 5779 FET cycles performed at the Reproductive and Genetic Center in the Affiliated Hospital of Shandong University of Traditional Chinese Medicine between January 2016 and December 2020. After propensity score matching, 3535 FET cycles were included, out of which 1238 were subjected to LAH while the remaining 2297 cycles were non-LAH (NLAH). The primary outcomes were clinical pregnancy rate (CPR) and live birth rate (LBR) while secondary outcomes included implantation rate (IR), biochemical pregnancy rate (BPR), ectopic pregnancy rate (EPR), pregnancy loss rate (PLR), multiple pregnancy rate (MPL), and monozygotic twinning rate (MTR). Logistic regression analysis was conducted to adjust for possible confounders. Subgroup analysis was also performed based on the endometrial preparation regimen.

Results

The LAH group exhibited a higher LBR, compared to the NLAH group (34.9% vs. 31.4%, OR = 1.185, 95% CI = 1.023, 1.374, P = 0.024). Additionally, the LAH group showed a decreasing trend in PLR and EPR; however, differences were insignificant (P = 0.078, P = 0.063 respectively). Differences in IR (24.6% vs. 24.3%), BPR (41.8% vs. 40.4%), CPR (40.7% vs. 38.3%), MPR (14.1% vs. 17.3%), and MTR (1.4% vs. 1.1%) were insignificant. Subgroup analysis revealed that LAH may be more conducive for pregnancy outcomes in hormone replacement cycles.

Conclusions

In summary, LAH has an increased chance of achieving live births. However, further prospective studies should be performed to confirm our findings.

冷凍解凍2次   vs.  冷凍解凍1次

冷凍解凍2次----仍會下降懷孕活產率 

. 2023 Oct;40(10):2357-2365.

 doi: 10.1007/s10815-023-02909-6. Epub 2023 Aug 16.

Evaluation of live birth rates and perinatal outcomes following two sequential vitrification/warming events at the zygote and blastocyst stages

Purpose: To study the outcome of sequential cryopreservation-thawing of zygotes followed by the cryopreservation-thawing of blastocysts in the course of an IVF treatment on live birth rate and neonatal parameters.

Methods: Single center, retrospective chart review for the time period of 2015-2020. Clinical and perinatal outcomes were compared between frozen embryo transfer cycles utilizing twice-cryopreserved (n = 182) vs. once-cryopreserved (n = 282) embryos. Univariate and multivariable analyses were used to adjust for relevant confounders.

Results: After adjustment for maternal age, gravidity, parity, body mass index (BMI), paternal age, fertilization method used, the number of oocytes retrieved in the fresh cycle, fertilization rate, and transfer medium, the transfer of twice-cryopreserved embryos resulted in a reduced probability of live birth (OR, 0.52; 95% CI 0.27-0.97; p=0.041) compared to once-cryopreserved embryos. No differences in the sex ratio, the mean gestational age, the mean length at birth, or the mean birth weight were found between the two groups.

Conclusion: The circumstantial use of sequential double vitrification-warming in course of treatment is associated with a reduced (but still reasonable) live birth rate compared to once-cryopreserved embryos. As the neonatal outcomes of twice-cryopreserved embryos are similar to once-cryopreserved embryos, this treatment option appears still valid as a rescue scenario in selected cases.

 IVF形成胚胎仍可施行胚胎切片& PGT

• 精子 DNA 在 Picoplex 和 ChromInst 條件下無法擴增，但可以使用 MDA 進行擴增。
• 在冷凍 PGT-A 週期中，cIVF-PGT-A 組和 ICSI-PGT-A 組之間每個週期的整倍體、嵌合體和非整倍體胚胎的平均速率沒有顯著差異。
• 夫妻接受 cIVF-PGT-A 治療的前瞻性研究結果顯示，在 150 個活檢滋養外胚層樣本中，沒有出現父源污染，只有 1 個母源污染。

• Sperm DNA failed to amplify under Picoplex and ChromInst conditions but could be amplified using MDA. 
• In frozen PGT-A cycles, no significant differences in the average rates of euploid, mosaic, and aneuploid embryos per cycle between the cIVF-PGT-A and ICSI-PGT-A groups were observed.
• The results of the prospective study that recruited couples for cIVF-PGT-A treatment showed no paternal contamination and one case of maternal contamination in 150 biopsied trophectoderm samples. 

. 2023 Oct;40(10):2333-2342.

 doi: 10.1007/s10815-023-02916-7. Epub 2023 Sep 1.

Conventional IVF is feasible in preimplantation genetic testing for aneuploidy

Purpose: To investigate the feasibility of the application of conventional in vitro fertilization (cIVF) for couples undergoing preimplantation genetic testing for aneuploidies (PGT-A) with non-male factor infertility.

Methods: To evaluate the efficiency of sperm whole-genome amplification (WGA), spermatozoa were subjected to three WGA protocols: Picoplex, ChromInst, and multiple displacement amplification (MDA). In the clinical studies, 641 couples who underwent PGT-A treatment for frozen embryos between January 2016 and December 2021 were included to retrospectively compare the chromosomal and clinical outcomes of cIVF and intracytoplasmic sperm injection (ICSI). Twenty-six couples were prospectively recruited for cIVF and PGT-A treatment between April 2021 and April 2022; parental contamination was analyzed in biopsied samples; and 12 aneuploid embryos were donated to validate the PGT-A results.

Results: Sperm DNA failed to amplify under Picoplex and ChromInst conditions but could be amplified using MDA. In frozen PGT-A cycles, no significant differences in the average rates of euploid, mosaic, and aneuploid embryos per cycle between the cIVF-PGT-A and ICSI-PGT-A groups were observed. The results of the prospective study that recruited couples for cIVF-PGT-A treatment showed no paternal contamination and one case of maternal contamination in 150 biopsied trophectoderm samples. Among the 12 donated embryos with whole-chromosome aneuploidy, 11 (91.7%) presented uniform chromosomal aberrations, which were in agreement with the original biopsy results.

Conclusions: Under the Picoplex and ChromInst WGA protocols, the risk of parental contamination in the cIVF-PGT-A cycles was low. Therefore, applying cIVF to couples with non-male factor infertility who are undergoing PGT-A is feasible.

PGT可以減少單次FET之流產率應無爭議

尤其針對年輕囊胚數量足夠狀況應優先考慮PGT 

但是否能提高整體累積懷孕率仍具爭議

甚至有研究顯示PGT減少整體累積懷孕率

尤其囊胚顆數不多之對象  是否需要增加暴露embryo biopsy之risk  仍具爭議

J Assist Reprod Genet. 2022 Nov; 39(11): 2547–2554.

Published online 2022 Sep 21. doi: 10.1007/s10815-022-02612-y

Does PGT-A improve assisted reproduction treatment success rates: what can the UK Register data tell us?

Purpose

To show how naïve analyses of aggregated UK ART Register data held by the Human Fertilisation and Embryology Authority to estimate the effects of PGT-A can be severely misleading and to indicate how it may be possible to do a more credible analysis. Given the limitations of the Register, we consider the extent to which such an analysis has the potential to answer questions about the real-world effectiveness of PGT-A.

Methods

We utilise the publicly available Register datasets and construct logistic regression models for live birth events (LBE) which adjust for confounding. We compare all PGT-A cycles to control groups of cycles that could have had PGT-A, excluding cycles that did not progress to having embryos for biopsy.

Results

The primary model gives an odds ratio for LBE of 0.82 (95% CI 0.68–1.00) suggesting PGT-A may be detrimental rather than beneficial. However, due to limitations in the availability of important variables in the public dataset, this cannot be considered a definitive estimate. We outline the steps required to enable a credible analysis of the Register data.

Conclusion

If we compare like with like groups, we obtain estimates of the effect of PGT-A that suggest an overall modest reduction in treatment success rates. These are in direct contrast to an invalid comparison of crude success rates. A detailed analysis of a fuller dataset is warranted, but it remains to be demonstrated whether the UK Register data can provide useful estimates of the impact of PGT-A when used as a treatment add-on.

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https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10504192/

Systematic review and meta-analysis: does pre-implantation genetic testing for aneuploidy at the blastocyst stage improve live birth rate?

Purpose

To establish if preimplantation genetic testing for aneuploidy (PGT-A) at the blastocyst stage improves the composite outcome of live birth rate and ongoing pregnancy rate per embryo transfer compared to conventional morphological assessment.

Methods

A systematic literature search was conducted using PubMed, EMBASE and Cochrane database from 1st March 2000 until 1st March 2022. Studies comparing reproductive outcomes following in vitro fertilisation using comprehensive chromosome screening (CCS) at the blastocyst stage with traditional morphological methods were evaluated.

Results

Of the 1307 citations identified, six randomised control trials (RCTs) and ten cohort studies fulfilled the inclusion criteria. The pooled data identified a benefit between PGT-A and control groups in the composite outcome of live birth rate and ongoing pregnancy per embryo transfer in both the RCT (RR 1.09, 95% CI 1.02–1.16) and cohort studies (RR 1.50, 95% CI 1.28–1.76). Euploid embryos identified by CCS were more likely to be successfully implanted amongst the RCT (RR 1.20, 95% CI 1.10–1.31) and cohort (RR 1.69, 95% CI 1.29–2.21) studies. The rate of miscarriage per clinical pregnancy is also significantly lower when CCS is implemented (RCT: RR 0.73, 95% CI 0.56–0.96 and cohort: RR 0.48, 95% CI 0.32–0.72).

Conclusions

CCS-based PGT-A at the blastocyst biopsy stage increases the composite outcome of live births and ongoing pregnancies per embryo transfer and reduces the rate of miscarriage compared to morphological assessment alone. In view of the limited number of studies included and the variation in methodology between studies, future reviews and analyses are required to confirm these findings.