雙重切片+雙重玻璃化冷凍 or 單次切片+雙重玻璃化冷凍解凍

vs.  單次切片+ 單次玻璃化冷凍解凍

----均導致活產率/持續懷孕率和臨床懷孕率下降。

Meta-Analysis

 

. 2024 Dec 1;39(12):2674-2684.

 doi: 10.1093/humrep/deae235.

Impacts of double biopsy and double vitrification on the clinical outcomes following euploid blastocyst transfer: a systematic review and meta-analysis

Study question: Compared to the 'single biopsy + single vitrification' approach, do 'double biopsy + double vitrification' or 'single biopsy + double vitrification' arrangements compromise subsequent clinical outcomes following euploidy blastocyst transfer?

Summary answer: Both 'double biopsy + double vitrification' and 'single biopsy + double vitrification' led to reduced live birth/ongoing pregnancy rates and clinical pregnancy rates.

What is known already?: It is not uncommon to receive inconclusive results following blastocyst biopsy and preimplantation genetic testing for aneuploidy (PGT-A). Often these blastocysts are warmed for re-test after a second biopsy, experiencing 'double biopsy + double vitrification'. Furthermore, to achieve better workflow, IVF laboratories may choose to routinely vitrify all blastocysts and schedule biopsy at a preferred timing, involving 'single biopsy + double vitrification'. However, in the current literature, there is a lack of systematic evaluation of both arrangements regarding their potential clinical risks in reference to the most common 'single biopsy + single vitrification' approach.

Study design, size, duration: A systematic review and meta-analysis were performed, with the protocol registered in PROSPERO (CRD42023469143). A search in PUBMED, EMBASE, and the Cochrane Library for relevant studies was carried out on 30 August 2023, using the keywords 'biopsy' and 'vitrification' and associated variations respectively. Only studies involving frozen transfers of PGT-A tested euploid blastocysts were included, with those involving PGT-M or PGT-SR excluded.

Participants/materials, setting, methods: Study groups included blastocysts having undergone 'double biopsy + double vitrification' or 'single biopsy + double vitrification', with a 'single biopsy + single vitrification' group used as control. The primary outcome was clinical pregnancy, while secondary outcomes included live birth/ongoing pregnancy, miscarriage, and post-warming survival rates. Random effects meta-analysis was performed with risk ratios (RR) and 95% CIs were used to present outcome comparisons.

Main results and the role of chance: A total of 607 records were identified through the initial search and nine studies (six full articles and three abstracts) were eventually included. Compared to 'single biopsy + single vitrification', 'double biopsy + double vitrification' was associated with reduced clinical pregnancy rates (six studies, n = 18 754; RR = 0.80, 95% CI = 0.71-0.89; I2 = 0%) and live birth/ongoing pregnancy rates (seven studies, n = 20 964; RR = 0.72, 95% CI = 0.63-0.82; I2 = 0%). However, no significant changes were seen in miscarriage rates (seven studies, n = 22 332; RR = 1.40, 95% CI = 0.92-2.11; I2 = 53%) and post-warming survival rates (three studies, n = 13 562; RR = 1.00, 95% CI = 0.99-1.01; I2 = 0%) following 'double biopsy + double vitrification'. Furthermore, 'single biopsy + double vitrification' was also linked with decreased clinical pregnancy rates (six studies, n = 13 284; RR = 0.84, 95% CI = 0.76-0.92; I2 = 39%) and live birth/ongoing pregnancy rates (seven studies, n = 16 800; RR = 0.79, 95% CI = 0.69-0.91; I2 = 70%), and increased miscarriage rates (five studies, n = 15 781; RR = 1.48, 95% CI = 1.31-1.67; I2 = 0%), but post-warming survival rates were not affected (three studies, n = 12 452; RR = 0.99, 95% CI = 0.97-1.01; I2 = 71%) by 'single biopsy + double vitrification'.

男性年齡& BMI與胚胎形態動力學 (PN~blastocyst時間點)

男性年齡與胚胎形態動力學早期階段（tPNa、tPNf、t2、t3、t4、t6）有顯著相關。

 male age was significantly associated with earlier embryo morphokinetics timings (tPNa, tPNf, t2, t3, t4, t6)

 tPNa time of pronuclei appearance, tPNf time of pronuclei fading

https://pmc.ncbi.nlm.nih.gov/articles/PMC12602791/

Impact of male factors on morphokinetic parameters: a prospective analysis using time-lapse monitored embryos

Introduction  Time-lapse technology enables recording embryo morphokinetic parameters, which are associated with embryonic competence and assisted reproductive technology (ART) outcomes. While female factors such as age and BMI are known to influence these parameters, the role of male factors remains understudied.

Aim  This study aimed to evaluate the influence of male factors on preimplantation embryo morphokinetics.

Methods  In this prospective observational study, 1,210 embryos from infertile couples undergoing Intracytoplasmic sperm injection (ICSI) or intracytoplasmic morphologically-selected sperm injection (IMSI) were monitored using time-lapse imaging. Male data, including age, BMI, sperm concentration, and sperm DNA fragmentation (SDF) were collected. Multiple regression analysis assessed the association between paternal factors and morphokinetic parameters, adjusting for female confounders.

Results   After adjustment, male age and BMI were found to significantly influence embryo developmental stages (from time to pronuclei appearance to t4 and t6 for age, from time to pronuclei appearance to t2 and t8 for BMI). The impact of sperm concentration was less consistent, and no significant relationship was observed with SDF.

Conclusions   These findings highlight the role of male factors, particularly age and BMI, in influencing embryo morphokinetics, even after accounting for female confounders. This underscores the potential for clinical interventions targeting paternal health to optimize ART outcomes. Additionally, the study reinforces the importance of considering both parental contributions in ART success, particularly the increasingly recognized influence of male age.

 GRHantagonist vs. progestin 誘導排卵COH

---GnRH antagonist可以取得較多卵子

Gonadotropin-releasing hormone antagonist protocol is associated with higher oocyte yield in young women at high risk for low oocyte retrieval: a retrospective study using three statistical methods

To study whether the gonadotropin-releasing hormone (GnRH) antagonist protocol is associated with improved oocyte retrieval outcomes compared with the progestin-primed ovarian stimulation (PPOS) protocol in young women at high predicted risk for low oocyte yield.

A total of 2,068 women aged ≤35 years undergoing their first in vitro fertilization and intracytoplasmic sperm injection treatment cycles between January 2023 and April 2025, identified as high risk for low oocyte retrieval (predicted score ≥0.41) using a validated nomogram incorporating factors such as age, antimüllerian hormone levels, antral follicle count, basal follicle-stimulating hormone levels, and follicle-stimulating hormone:luteinizing hormone ratio.

Exposure

Ovarian stimulation with either a GnRH antagonist protocol or a PPOS protocol, with protocol selection on the basis of clinical judgment and patient characteristics.

Main Outcome Measures

Incidence of low oocyte retrieval (<10 oocytes) and the number of oocytes retrieved.

Results

Among 2,068 young women at high predicted risk for low oocyte yield, the GnRH antagonist protocol resulted in significantly better ovarian stimulation outcomes compared with the PPOS protocol. After propensity score matching, the antagonist group had a higher mean number of oocytes retrieved (8.3 vs. 5.3; Bayes factor in favor of the alternative hypothesis [BF10] >1028) and a higher oocyte retrieval rate (88.2% vs. 81.2%; BF10 >103). The incidence of low oocyte retrieval (<10 oocytes) was significantly lower in the antagonist group (65.2% vs. 86.0%; BF10 >1019). No significant differences were observed in embryo quality or fertilization rates. Risk-based stratified analysis showed that the GnRH antagonist protocol significantly reduced the incidence of low oocyte retrieval in low-risk to midrisk groups (median odds ratio, 0.30–0.57, BF10 up to 647.8), whereas in the highest-risk subgroup, PPOS showed a potential advantage.

Conclusion

The GnRH antagonist protocol is superior to PPOS in maximizing oocyte retrieval among young women at high predicted risk for low oocyte retrieval, highlighting the value of individualized risk-based stimulation strategies to improve clinical outcomes.

• 常規 IVF 的 SCM 進行 niPGT 的方法與 ICSI 具有可比的性能
• 可擴大 niPGT 的應用範圍。

• 161個常規IVF胚胎和122個ICSI胚胎的niPGT表現無顯著差異（P > 0.05），
• IVF和ICSI方法的倍性一致率分別為75%和74.6%。
• niPGT胚胎的整倍體預測機率為 82.8%
• 性染色體嵌合體的預測機率為 77.8%，
• 低水平嵌合體的預測機率為 62.5%，
• 多條異常染色體的預測機率為 50.0%，
• 多條異常染色體的預測機率為 50.0

Observational Study

 

. 2022 Sep 4;20(1):396.

 doi: 10.1186/s12967-022-03596-0.

Non-invasive preimplantation genetic testing for conventional IVF blastocysts

Background: Previous studies suggested that non-invasive preimplantation genetic testing (niPGT) for intracytoplasmic sperm injection (ICSI) blastocysts can be used to identify chromosomal ploidy and chromosomal abnormalities. Here, we report the feasibility and performance of niPGT for conventional in vitro fertilization (IVF) blastocysts.

Methods: This was a prospective observational study. In the preclinical stage, whole genome amplification and NGS were performed using the sperm spent culture medium (SCM). Then, trophectoderm (TE) biopsies and corresponding SCM derived from 27 conventional IVF monopronuclear embryos were collected. In the clinical stage, samples from 25 conventional IVF cycles and 37 ICSI cycles from April 2020-August 2021 were collected for performance evaluation.

Results: Preclinically, we confirmed failed sperm DNA amplification under the current amplification system. Subsequent niPGT from the 27 monopronuclear blastocysts showed 69.2% concordance with PGT results of corresponding TE biopsies. In the clinical stage, no paternal contamination was observed in any of the 161 SCM samples from conventional IVF. While maternal contamination was observed in 29.8% (48/161) SCM samples, only 2.5% (4/161) samples had a contamination ratio ≥ 50%. Compared with that of TE biopsy, the performances of NiPGT from 161 conventional IVF embryos and 122 ICSI embryos were not significantly different (P > 0.05), with ploidy concordance rates of 75% and 74.6% for IVF and ICSI methods, respectively. Finally, evaluation of the euploid probability of embryos with different types of niPGT results showed prediction probabilities of 82.8%, 77.8%, 62.5%, 50.0%, 40.9% and 18.4% for euploidy, sex-chromosome mosaics only, low-level mosaics, multiple abnormal chromosomes, high-level mosaics and aneuploidy, respectively.

Conclusions: Our research results preliminarily confirm that the niPGT approach using SCM from conventional IVF has comparable performance with ICSI and might broadening the application scope of niPGT.

輔酶Q10是提高精子濃度最有效的治療方法

左旋肉鹼( Carnitine)是增強精子活力的最佳治療方法

Effects of Carnitine and Coenzyme Q10 on Sperm Quality and Pregnancy Rates in Men with Unexplained Infertility: A Systematic Review and Network Meta-Analysis

Aims

To study and explore the effects of oral Carnitine and Coenzyme Q10 on the quality of sperm and the rate of pregnancy in males with unexplained infertility.

Background

Preventing and reducing oxidative stress is essential for male infertility treatment. Antioxidants can effectively improve sperm quality and maintain sperm motility, thereby improving male fertility. At present, the most commonly used antioxidants in the clinic are carnitine and coenzyme Q10.

Methods

In this study, the effects of carnitine and coenzyme Q10 antioxidants on sperm quality and the rate of pregnancy in men with unexplained infertility were explored using a network meta-analysis method. The Cochrane Library, EMBASE, Web of Science and PubMed databases were searched for relevant studies. The observed indicators included semen concentration, progressive sperm motility, the rate of morphologically normal sperm and pregnancy rate.

Results

16 pieces of literature were included after rigorous screening. L-carnitine, coenzyme Q10 and L-carnitine + acetyl-L-carnitine significantly improved sperm quality parameters compared with placebo. CoQ10 induced the highest increase in sperm concentration (SMD 2.98 [95% CI: 1.13 to 7.87]). Furthermore, L-Carnitine has the greatest improvement effect on progressive sperm motility (SMD 4.19 [95% CI: 1.60 to 10.95]).

Conclusions

CoQ10 was the most effective intervention measure for improving sperm concentration, and L-carnitine was regarded as the best treatment method for enhancing sperm motility. In the future, we need more high-quality and large-sample studies to verify the effectiveness of antioxidants in the treatment of male infertility.

AMH水平與胚胎blastomere多核胚胎(MLN)的發生率有關

AMH水平每增加一個單位，MLN胚胎形成的風險就會增加1.12倍。

懷孕MLN胚胎/總胚胎的平均比率為0.34±0.18，

未懷孕MLN胚胎/總胚胎的平均比率為0.47±0.3（p=0.010）

Do serum AMH levels affect the incidence rate of multinucleated embryos in ICSI cycles?

A blastomere containing more than one nucleus is defined as a multinucleated blastomere. In our study, we aimed to investigate the relationship between serum anti-Mullerian hormone (AMH) levels and multinucleated (MLN) embryos, one of the parameters indicating embryo quality, in intracytoplasmic sperm injection (ICSI) cycles. The results of 888 ICSI cycles of patients aged 19–45 years attending an ART (assisted reproductive technology) clinic were retrospectively analysed. Cycles with at least one MLN embryo were defined as the study group (n = 237) and cycles without MLN embryos as the control group (n = 651). Univariate and multivariate logistic regression analyses were used to determine the risk factors affecting the dependent qualitative variables. The effect of AMH levels on multinucleation was found to be a significant risk factor (p < 0.001). One unit increase in AMH levels increases the risk of the presence of MLN embryos by 1.12 times. The mean MLN embryos/total embryo ratio in the group with clinical pregnancy was 0.34 ± 0.18, while the mean MLN embryos/total embryo ratio in the group that did not achieve clinical pregnancy was 0.47 ± 0.3 (p = 0.010). The presence of an MLN embryo has been associated with poor embryo development and ART outcomes. Parameters that can predict the formation of MLN embryos before treatment are crucial for the determination of the pregnancy rate. According to our results, serum AMH levels can be used as a predictive marker for the formation of MLN embryos.

 抗苗勒氏管激素 (AMH) 臨界值以確定多囊性卵巢形態

l      評估 AMH 臨界值 3.2 ng/mL 在經陰道超音波 (TVUS) 確定 PCOM 狀態的有效性。

l     根據血清AMH或陰道超音波(TVUS)檢測結果，計算PCOM陽性病例（PCOS表型為A、C或D的參與者）和陰性對照的數量。

l     3.2 ng/mL為AMH截斷值，   AMH 3.2 ng/m用於檢測PCOM的高準確性。在所有PCOM陽性的PCOS表型和體質指數分類中，AMH與PCOM狀態也具有一致性       

Original ArticleVolume 124, Issue 3p543-552  2025/9/162025

Anti-Müllerian hormone cutoff to determine polycystic ovarian morphology: HARMONIA study

• To verify the anti-Müllerian hormone (AMH) cutoff (3.2 ng/mL) for the determination of polycystic ovarian morphology (PCOM) as part of the latest polycystic ovary syndrome (PCOS) diagnosis in an independent prospective cohort of women of reproductive age.
• Assessment of the performance of the AMH cutoff of 3.2 ng/mL to identify PCOM status determined by transvaginal ultrasound (TVUS).
• The number of PCOM-positive cases (participants with PCOS phenotypes A, C, or D) and negative controls according to serum AMH or TVUS.
• Other measures included PCOM status within selected PCOS phenotypes and demographic/clinical factors including body mass index.
• Applying the AMH cutoff of 3.2 ng/mL, 118 of 128 cases and 639 of 820 controls were identified in accordance with TVUS (overall percent agreement, 79.9%).
• Agreement of AMH with PCOM status was also observed across all PCOM-positive PCOS phenotypes and body mass index categories.
• Conclusion
• The Elecsys AMH Plus immunoassay cutoff of 3.2 ng/mL was successfully verified, showing high accuracy to detect PCOM.
•  On the basis of current guideline recommendations for the detection of PCOM as part of PCOS diagnosis, these data support use of the AMH test as an easy-access diagnostic workup tool compared with TVUS.

 

 

 

Original ArticleVolume 124, Issue 3p468-477September 2025

Patients with a body mass index of ≥45 kg/m2 can safely undergo oocyte retrievals and anticipate similar assisted reproductive technology outcomes

比較體重指數 (BMI) 為 40-44.9 kg/m² 的患者與 BMI ≥45 kg/m² 的患者 的麻醉和輔助生殖技術 (ART) 結局。

l 本研究共納入 98 名 BMI ≥40 kg/m² 且接受取卵的患者：其中 BMI 在 40 至 44.9 kg/m² 之間的患者 56 名，BMI ≥45 kg/m² 的患者 42 名。

所有病人均成功接受靜脈鎮靜治療，無需更高程度的鎮靜或照護。

BMI ≥45 kg/m² 的患者平均手術時間比 BMI 40-44.9 kg/m² 的患者更長（26.8 分鐘 [標準差，13 分鐘] vs. 22.3 分鐘 [標準差，8.4 分鐘]）。

兩組在成熟卵母細胞獲獲數量、第 5/6 天囊胚數、整倍體胚胎數、臨床懷孕率、流產率或活產率均無差異。

使用靜脈鎮靜時，BMI ≥45 kg/m² 的患者與 BMI 40-44.9 kg/m² 的患者相比，其輔助生殖技術 (ART) 結局相似，且麻醉或輔助生殖技術併發症較少。

在適當的諮詢和術前準備下，BMI ≥45 kg/m² 的患者可以安全地進行取卵。

To compare anesthesia and assisted reproductive technology (ART) outcomes in patients with a body mass index (BMI) of 40–44.9 kg/m2 with those with a BMI of ≥45 kg/m2 because these patients are often excluded from care.

l           A total of 98 patients with a BMI of ≥40 kg/m2 undergoing oocyte retrieval were identified for the study: 56 patients with a BMI from 40 to 44.9 kg/m2 and 42 patients with a BMI of ≥45 kg/m2.

l           Demographics were not statistically significantly different between both groups, except that more patients with a BMI of 40–44.9 kg/m2 identified as White (73% vs. 60%) or Black (9% vs. 0).

l           All patients were successfully managed with intravenous sedation and did not require higher level of sedation or care.

l           The mean surgical duration was longer in patients with a BMI of ≥45 kg/m2 than in those with a BMI of 40–44.9 kg/m2 (26.8 minutes [standard deviation, 13 minutes] vs. 22.3 minutes [standard deviation, 8.4 minutes]).

l          There was no difference in the number of mature oocytes retrieved, day 5/6 blastocysts, the number of euploid embryos, or clinical pregnancy, miscarriage, or live birth rates.

l          When using intravenous sedation, patients with a BMI of ≥45 kg/m2 have similar ART outcomes with few anesthesia or ART complications compared with those with a BMI of 40–44.9 kg/m2.

l          With appropriate counseling and preoperative preparation, patients with a BMI of ≥45 kg/m2 can safely undergo oocyte retrieval.

使用progestin(PPOS, duphaston 20mg/d) vs  GnRH antagonist 應用於試管誘導排卵之LH抑制 

PPOS 組和 GnRH antagonist的整倍體率相當（分別為 12.5% 和 16.0%，P > 0.05）。

兩組FET在每次移植的妊娠試驗陽性率、臨床懷孕率、流產率、子宮外孕率或活產率方面均無顯著差異。

Comparison of the euploidy rate  following progestin-primed vs GnRHantagonist protocol

• Euploidy rate was comparable between the PPOS and antagonist group (12.5% vs. 16.0% respectively, P > 0.05). 
• No significant differences were observed between the two groups in positive pregnancy test, clinical pregnancy, miscarriage, ectopic pregnancy, or live birth rates per transfer in the first frozen embryo transfer cycles.
• Both PPOS and antagonist protocols had similar euploidy rates in PGT-A cycles.

Variables	PPOS group (n = 120)	Antagonist group (n = 120)	P-value
Starting dose of FSH (IU)	225.0 (225.0-225.0)	225.0 (225.0-225.0)	0.134
Total dosage of FSH (IU)	1800.0 (1575.0-2025.0)	1800.0 (1575.0-2212.5)	0.010
Duration of stimulation (days)	8.0 (7.0-8.8)	8.0 (7.0–9.0)	0.053
Serum estradiol level on trigger day (pg/ml)	1870.5 (912.7-2940.1)	1061.3 (592.0-2614.1)	0.007
Serum LH level on trigger day (IU/l)	4.3 (2.7–5.9)	2.4 (1.6–3.6)	0.000
Serum progesterone level on trigger day (ng/ml)	0.9 (0.6–1.2)	0.8 (0.6–1.2)	0.588
Premature ovulation (%)	0.8 (1/120)	0.8 (1/120)	1
No. of retrieved oocytes (n)	6.0 (3.0–10.0)	5.0 (3.0–10.0)	0.914
No. of mature occytes (n)	4.5 (2.0–8.0)	4.0 (3.0–9.0)	0.980
No. of oocytes fertilized (n)	3.0 (2.0–7.0)	3.0 (2.0–6.0)	0.728
Fertilization rate (%)	94.4 (75.0-100)	92.9 (75.0-100)	0.746
Cleavage rate (%)	100 (100–100)	100 (100–100)	0.648
No. of blastocysts formation (n)	1.5 (0–3.0)	1.0 (0–3.0)	0.422
Blastocysts formation rate (%)	59.4 (33.3–100)	50.0 (33.3–80)	0.299
Total No. of euploid blastocysts	93	97
No. of euploid blastocysts (n)	0 (0–1.0)	0 (0–1.0)	0.995
Euploid blastocysts rate per injected oocyte (%)	12.5 (0–25.0)	16.0 (0-27.7)	0.477
Euploid blastocysts rate per woman (%)	33.3 (0-66.7)	50.0 (0-66.7)	0.459
No. of cycles with no blastocyst for biopsy (%)	27.5 (33/120)	30.0 (36/120)	0.669
No. of cycles with no euploid blastocysts for transfer (%)	52.5 (63/120)	55.0 (66/120)	0.698

PPOS group (n = 53)	Antagonist group (n = 47)	P-value
Endometrial preparation, n (%)			0.995
Natural cycles	1.9 (1/53)	2.0% (1/47)
Clomid-induced	11.5 (6/53)	10.0 (5/47)
Hormonal cycles	83.0 (44/53)	91.5 (43/47)
Endometrial thickness (day of trigger) (mm)	10.0 (8.6–11.0)	9.1 (8.5–10.4)	0.600
hCG test positive rate (%)	66.0 (35/53)	70.2 (33/47)	0.655
Clinical pregnancy rate (%)	58.5 (31/53)	59.6 (28/47)	0.912
Clinical miscarriage rate (%)	9.7 (3/31)	17.9 (5/28)	0.359
Ectopic pregnancy rate (%)	0 (0/35)	0 (0/33)	0
Live birth rate (%)	52.8 (28/53)	48.9 (23/47)	0.359

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