中空透明帶(ZP)之偏光折射率(birefringence)較高----卵子品質較好

Analysis of correlations between zona pellucida birefringence and molecular markers of oocyte developmental competence

BMC Genomics volume 15, Article number: P48 (2014) 

• 
  
• Zona Pellucida (ZP) , a filamentous matrix of glycosylated glycoproteins surrounding the oocyte
• In fact, ZP thickness and birefringence was reported to be positively correlated with higher ability of the oocyte to achieve successful pregnancy,
• We expect to find a positive correlation between the ZP birefringence and molecular markers of oocyte competence.  
• ZP thickness and birefringence was reported to be positively correlated with higher ability of the oocyte to achieve successful pregnancy
• 
  
  
  

 卵子大小(直徑)會隨年紀逐漸下降

中空透明帶厚度也會隨年紀逐漸下降

Correlation of oocyte morphometry parameters with woman’s age

• April 2011
• Journal of Assisted Reproduction and Genetics 28(6):545-52

最理想ㄉ卵子直徑為100-120um

太大太小都會影響懷孕率

Reproductive Sciences volume 28, pages904–908 (2021)

Can Oocyte Diameter Predict Embryo Quality?

• whether oocyte diameter might predict the quality of the developing embryo. A retrospective, single-center cohort study. 
• Oocytes were grouped by mean oocyte diameter (MOD) and incubated for 5 days before the final morphological evaluation done by an expert embryologist. 
• A total of 471 cycles which yielded 3355 metaphase II oocytes were included in the analysis. 
• Embryos developed from oocytes with MOD close to the average (Average 1SD < MOD < Average + 1SD) had increased good-quality blastulation rates compared with embryos that developed from very small or very large oocytes. 
• Oocytes with MOD between 105.96 and 118.69 μm have better probability of becoming top-quality D5 blastocysts (17.1-17.4% grade 1 embryos). 

卵子與卵丘顆粒 & theca細胞之間有相當複雜之訊號&賀爾蒙交互作用

講解https://www.youtube.com/watch?v=2sn2-7EfLMw

J Assist Reprod Genet. 2022 May; 39(5): 1011–1025.

Published online 2022 May 2. doi: 10.1007/s10815-022-02438-8

Mammalian cumulus-oocyte complex communication: a dialog through long and short distance messaging

• Communications are crucial to ovarian follicle development and to ovulation, and while both folliculogenesis and oogenesis are distinct processes, they share highly interdependent signaling pathways. 
• Signals from distant organs such as the brain must be processed and compartments within the follicle have to be synchronized. 
• The hypothalamic–pituitary–gonadal (HPG) axis relies on long-distance signalling analogous to wireless communication by which data is disseminated in the environment and cells equipped with the appropriate receptors receive and interpret the messages. 
• In contrast, direct cell-to-cell transfer of molecules is a very targeted, short distance messaging system. 

胚胎植入後子宮內膜是否增厚與懷孕與否無明顯相關

Endometrial thickness changes after progesterone administration do not affect the pregnancy outcomes of frozen-thawed euploid blastocyst transfer: a retrospective cohort study

https://www.fertstert.org/action/showPdf?pii=S0015-0282%2821%2901931-2

• In the euploid blastocyst transfer cycle of preparing the endometrium with HRT, the endometrial thickness change ratio after progesterone administration was not related to pregnancy outcomes. 
• Some scholars believe that when the endometrial thickness is <7 mm or 8 mm before ovulation, the cycle should be canceled because of the low clinical pregnancy rate and the high miscarriage rate (7–9). 
• Some studies have shown that the endometrial thickness at the end of proliferation ranges from 9 to 14 mm to achieve the best implantation rate and clinical pregnancy rate (10)
• The study by Bu et al. (19) showed that regardless of which endometrial preparation protocol was used, endometrial increases after progesterone administration had a higher clinical pregnancy rate. 

 PRP 血小板濃縮血漿  可用於刺激子宮內膜厚度  下降流產率  提高卵巢功能

詳細解說

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8190208/

J Assist Reprod Genet. 2021 May; 38(5): 1003–1012.

Published online 2021 Mar 15. doi: 10.1007/s10815-021-02146-9

A narrative review of platelet-rich plasma (PRP) in reproductive medicine

• PRP is autologous blood plasma that has been enriched with platelets at about 4-5 times more than the circulating blood. PRP can stimulate proliferation and regeneration with a large amount of growth factors and cytokines, including PDGF, TGF, VGEF, EGF, fibroblast growth factor (FGF), insulin-like growth factor I, II (IGF I, II), interleukin 8 (IL-8) and connective tissue growth factor (CTGF). 
• Currently, PRP infusion is being increasingly used in several fields in medicine such as nerve injury, osteoarthritis, chronic tendinitis, bone repair and regeneration, cardiac muscles, alopecia, plastic surgery and oral surgery, but there is limited experience in gynecology and obstetrics
• In women with thin endometrium, the literature shows an increase in endometrial thickness and increase in chemical and clinical pregnancy rates following autologous PRP therapy. 
• In women with poor ovarian reserve, autologous intraovarian PRP therapy increased anti-Mullerian hormone (AMH) levels and decreased follicle-stimulating hormone (FSH), with a trend toward increasing clinical and live birth rates. 
• This trend was also noted in women with recurrent implantation failure.
• Limited literature shows promise in increasing endometrial thickness, increasing AMH, and decreasing FSH levels, as well as increasing chemical and clinical pregnancy rates. 
• The lack of standardization of PRP preparation along with the lack of large randomized controlled trials needs to be addressed in future studies. 

• the methods of preparation for PRP also lacked consistency. 
• Kamath et al. discuss using 0.5 to 1.0 ml of PRP for intrauterine infusion; however, they do not discuss the method of preparation or if a platelet agonist was used to activate the platelets and if so, which agonist was used [19]. 
• There are different types of platelet agonists used for activation, including calcium chloride, thrombin, and collagen [20]. 
• Chang et al. collected 15ml of whole blood from the participants and then used the two-step centrifugation process, the initial at 300 × g for 10 min at 18°C which results in 3 layers consisting of red blood cells at the bottom, buffy coat in the middle, and cellular plasma in the supernatant [21]. 
• The top two layers were placed in a separated tube and centrifuged at 700 × g for 15 min at 18°C, and 0.5 to 1.0 ml of PRP was pipetted out for infusion purposes [21]. 
• Coksuer et al. collected the sample at room temperature and centrifuged at 1500 × g for 5 min, the supernatant was then removed from the tube which was then recapped and underwent inversion/resuspension, and the remaining sample (1 ml of PRP) was utilized for infusion [5].
• Nazari et al. centrifuged the sample immediately after collection initially at 1200 rpm for 10 min and then centrifuged again at 3300 rpm for 5 min to collect 0.5ml of PRP for infusion [9]. 
• PRP was prepared from autologous blood using a two-step centrifuge process. On the 9th or 10th day of the mensural cycle, 17.5 ml of peripheral venous blood was drawn in the syringe that contained 2.5 ml of Acid Citrate A Anticoagulant solution (ACD-A) (Arya Mabna Tashkhis, Iran) and centrifuged immediately at 1200 rpm for 12 min to separate the red blood cells. 
• The plasma was centrifuged again at 3300 rpm for 7 min to obtain the PRP. Then, 0.5 ml of PRP was infused into the uterine cavity with the IUI catheter (Takwin, Iran).
• Three months after undergoing intraovarian autologous PRP injection, there was an overall decrease in FSH by 67.33% and an increase in AMH by 75.18% 

 冷凍卵子 vs  新鮮卵子  懷孕率類似

冷凍卵子受損率約10-20%

卵子捐贈者之年齡是影響冷凍結果之關鍵

https://academic.oup.com/biolreprod/article/106/2/316/6485190?login=false

• Though oocytes can be cryopreserved without ice crystal formation through vitrification, it is still not clear whether the process of vitrification causes any negative impact (temperature change/chilling effect, osmotic stress, cryoprotectant toxicity, and/or phase transitions) on oocyte quality that translate to diminished embryo developmental potential or subsequent clinical outcomes. 
•  Currently, the most used human oocyte vitrification method is using the combination of EG and DMSO in order to achieve the lowest concentrations required from each individual CPA [8, 10, 12, 14, 25].
• The authors concluded that outcomes obtained with vitrified oocytes are as good as those with fresh oocytes [15].
• most of the oocyte vitrification studies lead us to conclude that the developmental capability of embryos obtained from vitrified oocytes is not affected by the vitrification procedure, as fertilization, embryo cleavage, embryo quality, and clinical results were similar to those achieved with fresh oocytes. 
• a portion of the vitrified oocytes did not survive the vitrification and warming process (~20%), 
• The results show that fertilization and early embryo development (defined by day 3 good-quality embryo rates and blastocyst formation rates) were not statistically significantly different in the vitrified oocytes compared with the fresh sibling oocytes. 
• These results strongly imply that the potential negative impact of vitrification on oocytes can be minimized. 
• The oocyte spindle has shown rapid depolymerization when exposed to sub-physiological temperatures [86, 88]
• the time interval between each cell division was not changed, indicating that DNA replication was not disrupted in the embryos coming from vitrified oocytes. 
• Despite this statistically significant delay the time to blastulation, the duration of the cell cycle was still consistent with fresh counterparts [127, 128]
• It remains unknown whether the process of vitrification causes subtle damage 

胚胎整個生長過程PH值並非一成不變

• 受孕期PH值偏鹼7.4-7.4.5
• 分裂期間PH值偏酸7.2-7.3
• 桑葚期囊胚期PH值偏鹼7.25-7.35
• 子宮腔PH值偏酸PH<7.1

 https://academic.oup.com/humupd/article/18/3/333/608432?login=false

Is there an optimal pH for culture media used in clinical IVF? 

J.E. Swain

Human Reproduction Update, Volume 18, Issue 3, May/June 2012, Pages 333–339, https://doi.org/10.1093/humupd/dmr053

BACKGROUND

Reducing environmental stress imposed upon gametes and embryos in the IVF laboratory is crucial in optimizing culture conditions and development. One environmental parameter of particular importance is external pH (pHe) of culture media. An optimal pHe has not been identified.

METHODS

Electronic searches were performed using keywords focused on pH and the embryo using PUBMED through August 2011, with no limits placed on a beginning time. Examples of keywords include CO₂, bicarbonate and hydrogen ion. Relevant papers were then examined to obtain additional publications.

RESULTS

Determining optimal pHe is problematic due to difficulty in isolating pHe from other variables, such as CO₂ and bicarbonate. Various commercial media companies recommend differing pHe ranges, most within the range of 7.2–7.4, with some companies recommending altering pHe based on the gamete or stage of the embryo. However, changing pHe during culture has not been experimentally shown to improve outcomes. Further complicating attempts to define an optimal pHe is that media components can impact intracellular pH (pHi). As a result, media with different concentrations of substances, such as lactate or amino acids, may have different pHi, despite being in the same pHe.

CONCLUSIONS

Due to the plasticity of embryos, a range of pHe's can support development, and defining an optimal pHe is difficult. It is unclear whether there is any benefit in changing pHe at various steps during IVF. The ideal pHe will likely vary from media to media and, until comparative studies have been performed isolating pHe, adherence to manufacturer recommendations and maintenance of a small acceptable pHe range are advisable. 

• 
  
• What is known is that the pHi of embryos is ∼7.1–7.2. 
•  pHe should be slightly higher than pHi to help offset the acidification that occurs as a result of intracellular metabolic processes. Thus, commercial media manufacturers often recommend, and many laboratories culture their embryos, in the range of pHi 7.2–7.4, 
• the use of a ‘high-low-high’ system, where pHe is elevated for fertilization, lowered for cleavage-stage embryo culture and again raised for blastocyst culture has been promoted by commercial media manufactures 
•  better sperm penetration at pHe 7.6–7.8, 
• pHe of the oviduct in species like the monkey, rabbit and bovine appears to have a more alkaline pHe (∼7.5–7.9) (Iritani et al., 1971; Maas et al., 1977; Hugentobler et al., 2004), 
• pHe of human uterine fluid has been measured with a pH electrode and found to be ∼6.6, while another study found a pHe of ∼7.1 (Sedlis et al., 1967; Yedwab et al., 1976).
•  a ‘high-high-low’ or ‘high-low-low’ scheme may be more suitable/physiologic for fertilization, cleavage and blastocyst development.

 囊胚期胚胎  女性囊胚比例略高 54%

女性囊胚染色體異常率較高

著床率  男女無差異

Day5囊胚  著床率高於  Day6囊胚

Embryo quality, ploidy, and transfer outcomes in male versus female blastocysts

Journal of Assisted Reproduction and Genetics volume 38, pages2363–2370 (2021)

Purpose

The goal is to determine if variations exist between male and female blastocysts in preimplantation measurements of quality and ploidy and in vitro fertilization elective single-embryo transfer (eSET) outcomes.

Methods

A retrospective chart review was conducted from a private fertility center’s database of blastocysts undergoing preimplantation genetic testing for aneuploidy, along with details of eSET from this screened cohort. Main outcomes included preimplantation embryo quality and sex-specific eSET outcomes.

Results

A total of 3708 embryos from 578 women were evaluated, with 45.9% male and 54.1% female. The majority were High grade. No difference existed between embryo sex and overall morphological grade, inner cell mass or trophectoderm grade, or blastocyst transformation day. Female blastocysts had a higher aneuploidy rate than male blastocysts (P < 0.001). Five hundred thirty-nine eSETs from 392 women were evaluated, with High grade embryos more likely to have implantation (P < 0.001), clinical pregnancy (P < 0.001), and ongoing pregnancy (P = 0.018) than Mid or Low grade embryos. Day 5 blastocysts were more likely to have implantation (P = 0.018), clinical pregnancy (P = 0.005), and ongoing pregnancy (P = 0.018) than day 6 blastocysts. Male and female embryos had similar transfer outcomes, although female day 5 blastocysts were more likely to result in clinical pregnancy (P = 0.012), but not ongoing pregnancy, than female day 6 blastocysts. Male eSET outcomes did not differ by blastocyst transformation day.

Conclusion

Male and female embryos have comparable grade and quality; however, female embryos were more likely to be aneuploid. Ongoing pregnancy rates did not differ by embryo sex. Day 5 embryos had more favorable transfer outcomes than day 6 embryos.

 囊胚期胚胎  男性囊胚比例略高 53%

男性囊胚染色體異常率較高

著床率  男女無差異

Male and female blastocysts: any difference other than the sex?https://www.rbmojournal.com/article/S1472-6483%2822%2900347-9/fulltext# 

Research question

Is there any imbalance in the sex ratio at the blastocyst stage of human embryos? And what is the sex ratio in euploid, transferred, implanted blastocysts and at birth?

Design

Embryos from 646 women undergoing 921 preimplantation genetic testing for aneuploidy (PGT-A) cycles from September 2017 to February 2020 were included. Data from the chromosomal constitution of 2637 biopsied blastocysts were retrospectively analysed. Trophectoderm samples were analysed by next-generation sequencing. Embryos were categorized as euploid, mosaic or aneuploid. A total of 548 blastocysts diagnosed as euploid were warmed and transferred in a subsequent single-embryo transfer cycle.

Results

The blastocyst sex ratio was skewed in favour of male sex with 53.1% (1401/2637) of blastocysts diagnosed as male and 46.9% (1236/2637) as female (sex ratio 1.13, 95% confidence interval [CI] 1.05–1.22). Following biopsy and PGT-A, 41.2% (1086/2637) of blastocysts were classified as euploid, 7.7% (202/2637) as mosaic and 51.2% (1349/2637) as aneuploid. More chromosome euploidy was observed among female than male blastocysts (adjusted odds ratio 1.29, 95% CI 1.08–1.55) after adjusting for female age, male age and gonadotrophin dose. Euploid blastocysts were comparable between the sexes (sex ratio 0.99, 95% CI 0.88–1.11). No significant differences were observed between the sexes in implantation (sex ratio 0.86, 95% CI 0.68–1.08), miscarriage (sex ratio 1, 95% CI 0.51–1.97) or live birth rate (sex ratio 0.85, 95% CI 0.66–1.08).

Conclusions

More male than female embryos develop to the blastocyst stage. Male blastocysts exhibit a higher aneuploidy rate. The capacity to implant and lead to a live birth is similar between the sexes.

 primary ciliary dyskinesia (PCD, 原发性纤毛运动障碍) 

• 基因異常導致
• 跟不孕症有相當關連  尤其是男性不孕症
• 跟女性不孕症有相當關連  主要影響輸卵管內膜纖毛功能
• 影響呼吸道纖毛功能

• more than 40 genes being identified in over 70% of patients with PCD (Lucas et al., 2020)

• The exact prevalence of subfertility in PCD is unclear but appears to be higher in men (up to 83% affected) compared to women (up to 61% affected). 

https://academic.oup.com/humupd/advance-article/doi/10.1093/humupd/dmad003/7017573?searchresult=1

The impact of primary ciliary dyskinesia on female and male fertility: a narrative review 

Human Reproduction Update, dmad003, https://doi.org/10.1093/humupd/dmad003

Published:

 

31 January 2023

 Article history

BACKGROUND

Primary ciliary dyskinesia (PCD) is a genetic condition affecting the structure and function of sperm flagellum and motile cilia including those in the male and female reproductive tracts. Infertility is a commonly reported feature of PCD, but there is uncertainty as to how best to counsel patients on their fertility prognosis.

OBJECTIVE AND RATIONALE

This review aimed to summarize the prevalence of subfertility, possible underlying mechanisms, and the success of ART in men and women with PCD. The efficacy of ART in this patient group is relatively unknown and, hence, the management of infertility in PCD patients remains a challenge. There are no previous published or registered systematic reviews of fertility outcomes in PCD.

SEARCH METHODS

Systematic literature searches were performed in Medline, Embase, Cochrane Library, and PubMed electronic databases to identify publications between 1964 and 2022 reporting fertility outcomes in men and women with PCD. Publications were excluded if they reported only animal studies, where gender was not specified or where subjects had a medical co-morbidity also known to impact fertility. Quality of evidence was assessed by critical appraisal and application of an appraisal tool for cross-sectional studies. The primary outcomes were natural conception in men and women with PCD, and conception following ART in men and women with PCD.

OUTCOMES

A total of 1565 publications were identified, and 108 publications were included after screening by two independent researchers. The quality of available evidence was low. The exact prevalence of subfertility in PCD is unclear but appears to be higher in men (up to 83% affected) compared to women (up to 61% affected). Variation in the prevalence of subfertility was observed between geographic populations which may be explained by differences in underlying genotype and cilia function. Limited evidence suggests subfertility in affected individuals is likely caused by abnormal cilia motion in the fallopian tubes, endometrium and efferent ductules, and dysmotile sperm. Some men and women with PCD benefited from ART, which suggests its use should be considered in the management of subfertility in this patient group. Further epidemiological and controlled studies are needed to determine the predictors of fertility and optimal management in this patient group.