Search

Research question To assess the relationship between the number of oocytes retrieved during elective oocyte cryopreservation (EOC) cycles with various clinical, biochemical, and radiological markers, including age, body mass index (BMI), baseline anti-Müllerian hormone (AMH), antral follicle count (AFC), Oestradiol level (E2) and total number of follicles ≥ 12 mm on the day of trigger. To also report the reproductive outcomes from women who underwent EOC. Methods A retrospective cohort of 373 women embarking on EOC and autologous oocyte thaw cycles between 2008 and 2018 from a single London clinic in the United Kingdom. Results 483 stimulation cycles were undertaken amongst 373 women. The median (range) age at cryopreservation was 38 (26–47) years old. The median numbers of oocytes retrieved per cycle was 8 (0–37) and the median total oocytes cryopreserved per woman was 8 (0–45). BMI, E2 level and number of follicles ≥ 12 mm at trigger were all significant predictors of oocyte yield. Multivariate analysis confirmed there was no significant relationship between AFC or AMH, whilst on univariate analysis statistical significance was proven. Thirty six women returned to use their cryopreserved oocytes, of which there were 41 autologous oocyte thaw cycles undertaken. There were 12 successful livebirths achieved by 11 women. The overall livebirth rate was 26.8% per cycle. No livebirths were achieved in women who underwent EOC ≥ 40 years old, and 82% of all livebirths were achieved in women who had done so between 36 and 39 years old. Conclusion Clinical, biochemical and radiological markers can predict oocyte yield in EOC cycles. Reproductive outcomes are more favourable when cryopreservation is performed before the age of 36, with lower success rates of livebirth observed in women aged 40 years and above.

PurposePre-implantation genetic testing for aneuploidies (PGT-A) is a technique used as part of in vitro fertilisation to improve outcomes. Despite the upward trend in women utilising PGT-A, data on women’s motivations and concerns toward using the technology, and perceptions having undergone the process, remain scarce.MethodsThis cross-sectional survey, based at a fertility clinic in the UK, utilised an electronic questionnaire to assess the motivations of women who undergo PGT-A and their perceptions and attitudes toward PGT-A after using it.ResultsOne hundred sixty-one women responded. The most significant motivating factors to undergo PGT-A were to improve the probability of having a baby per cycle (9.0 ± 2.1) and enhance the chance of implantation (8.8 ± 2.5). The least important motivations were reducing the number of embryos transferred per cycle (2.7 ± 3.3) and saving money by reducing the number of procedures required (4.6 ± 3.4). The most significant concerning factors identified included not having embryos to transfer (5.7 ± 3.4) and the potential for embryo damage (5.2 ± 3.3). The least concerning factors included religious (0.6 ± 1.7) or moral (1 ± 2.2) concerns. The majority of women were satisfied/very satisfied following treatment (n= 109; 68%). The proportion of those who were satisfied/very satisfied increased to 94.2% (n= 81) following a successful outcome, and reduced to 43.5% (n= 27) in those who had an unsuccessful outcome or had not undergone embryo transfer (p< 0.001).ConclusionThis study highlights that perceptions amongst women who use PGT-A are mostly positive. We also demonstrate a significant association between satisfaction and reproductive outcomes, with those who achieve a live birth reporting more positive perceptions toward PGT-A.

There is accumulating evidence demonstrating that positive lifestyle modification and the optimization of the preconceptual period can influence the reproductive potential for both men and women. However, a large percentage of couples attending fertility clinics with potential to improve preconception habits may not always receive appropriate preconceptual advice. In addition, supplements and adjuncts that promise to increase fertility treatment success rates are marketed to infertile patients despite lack of convincing evidence supporting their benefit. This review aims to identify possible associations between lifestyle factors for couples seeking fertility treatment and fertility treatment outcomes and to offer possible explanations of the biological basis of these associations. An electronic search was conducted from 1978 through July 2019 linking preconceptual behaviors for women and men with the outcome of fertility treatment. The literature search explored the importance of numerous factors, including smoking, caffeine, alcohol, obesity, physical exercise, recreational drugs, stress, diet, supplements, alternative medicine, environmental factors, and pollutants. Some associations were found to be more significant than others. The preconceptual period is undeniably a delicate and important window which should not be overlooked during fertility counseling. Simple lifestyle modifications could positively influence fertility treatment outcomes. Fertility teams, consisting of clinicians, fertility nurses, dieticians, psychologists, exercise advisors and others, should dedicate time to offer evidence-based preconceptual advice and targeted interventions to couples seeking fertility treatment.

A delay in childbearing to later in life has increased the number of women of advanced maternal age (AMA) opting for assisted reproduction. Women should be made aware that there are age-related changes to fertility, including a decline in oocyte reserve and quality, in addition to an increase in the number of oocyte chromosomal aberrations. Success rates of assisted reproductive technology (ART) cycles decrease with advanced maternal age. There are different fertility options for women of AMA, including fertility preservation (oocyte or embryo freezing), in vitro fertilisation (IVF treatment) with or without preimplantation genetic screening and oocyte or embryo donation. Detailed counselling needs to be offered to these women with regard to the risks, success rates, ethical and legal implications of these fertility treatment options. Women of AMA should be screened for underlying medical conditions that could have an impact on maternal and neonatal morbidity and mortality.

To determine if oocyte yield in women undergoing cryopreservation for social (SOC), medical (MOC) and oocyte donation (OD) cycles is comparable when matched for age. 315 oocyte retrievals were performed for SOC, 116 for MOC and 392 for OD. Non-parametric Kruskal-Wallis tests and Poisson regression were used to assess the impact of age stratification. The median ages of women undergoing SOC, MOC, and OD were 38, 31 and 26 years respectively. The median (IQR) number of oocytes in the three categories was 7, 10, and 12. The oocyte yield was significantly higher in women aged 30-34 years undergoing SOC, compared to the MOC group. For the SOC group, age in years, oestradiol levels per 1000 pmol/and follicle count12mm on the day of trigger were significant predictors of oocyte yield. Women embarking on SOC are significantly older than those undergoing MOC and OD, and thus oocyte yield is reduced when stratified for age. This study highlights the significant predictors of oocyte yield amongst women undergoing oocyte cryopreservation for specific indications. The findings can be used to optimise the yield and overall chance of successful livebirth.

Objective To investigate whether paternal age exerts an independent effect on the clinical outcomes of assisted reproductive technology (ART) cycles. Evidence Review Observational studies were identified through a systematic search of MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, and National Health Service evidence. Data for women aged ≤39 years were extracted and analyzed. We included all studies, both autologous and donor oocyte, into separate analyses of the effect of paternal age on ART outcome. We excluded studies in azoospermic men, women aged ≥40 years, ART including preimplantation genetic testing, and involving donor sperm. The included studies scored well on the Newcastle-Ottawa Quality Assessment Scale for observational studies. The primary outcome was live birth rate and secondary outcome measures were clinical pregnancy rate and miscarriage rate. When pooling data, the random-effects model was used to counter the effect of heterogeneity in the studies. Result(s) Live birth rate was reported in three autologous oocyte studies (2,926 cycles) and five donor oocyte studies (7,648 cycles). Live birth rate was found to be increased significantly when male age was 40 years in autologous studies. In donor oocyte studies, the miscarriage rate was not increased when male age was >50 years. All of the autologous oocyte studies reported a statistically significant association between male age and female age. Conclusion(s) The findings of this review and meta-analysis, based on the donor oocyte model, suggest that advanced paternal age does not exert an independent effect on the outcome of ART cycles. Miscarriage rates do not appear to be increased even for men >50 years of age after treatment with donor oocytes. The meta-analysis of autologous oocyte studies suggests that increasing male age may have a deleterious effect on the outcome of ART, however, this may be confounded by the strong association with increasing maternal age.

PURPOSE: To compare reproductive outcomes following a euploid embryo transfer, between those embryos vitrified-warmed twice to those vitrified-warmed once. METHODS: We retrospectively analysed 694 single euploid frozen embryo transfer cycles following preimplantation genetic testing for aneuploidy (PGT-A). For cycles in group 1 (N = 451), embryos were biopsied for PGT-A at blastocyst stage and vitrified. For cycles in group 2 (N = 146), embryos were vitrified at blastocyst stage, before being warmed and biopsied for PGT-A and vitrified again. For cycles in group 3 (N = 97), embryos were vitrified on day-3, before being warmed, cultured to day-5 and biopsied for PGT-A and re-vitrified. RESULTS: The pregnancy, clinical pregnancy and livebirth rate in group 2 were not statistically different to group 1 (pregnancy rate, adjusted OR 1.09, 95% CI 0.62–1.91; clinical pregnancy, aOR 0.89, 95% CI 0.58–1.37; live birth rate, aOR 0.85, 95% CI 0.56–1.28). There was also no significant difference between group 3 and group 1, with similar pregnancy rate (aOR 1.22, 95% CI 0.74–1.99), clinical pregnancy rate (aOR 1.21, 95% CI 0.75–1.96) and live birth rate (aOR 1.15, 95% CI, 0.73–1.80). There was no significant difference in miscarriage rates between all three groups. The age at the oocyte collection, embryo quality and day of biopsy were associated with pregnancy, clinical pregnancy and live birth rate. CONCLUSION: This study suggests that vitrifying and warming embryos twice at blastocyst or at cleavage and then blastocyst stage, can lead to similar reproductive outcomes to embryos vitrified-warmed once, after a single euploid embryo transfer.

IntroductionAdenomyosis can adversely reduce chances of pregnancy in couples undergoing assisted conception. We aim to evaluate the effect of two different downregulation protocols on the reproductive outcomes in women with moderate and severe adenomyosis undergoing frozen-thawed embryo transfer (FTET).Methods and analysisWe will conduct a two-armed pragmatic randomised clinical trial comparing modified downregulation with gonadotrophin-releasing hormone (GnRH) analogue for 6 weeks to standard downregulation with GnRH analogue for 1 week prior to FTET. Our primary outcome is clinical pregnancy, defined as a viable intrauterine pregnancy confirmed by ultrasound at greater than 6 weeks gestation, with other secondary reproductive, neonatal and safety outcomes. We aim to randomise 162 patients over 3 years to achieve 80% power for detecting a 20% difference in the primary outcome at 5% significance.Ethics and disseminationTo date there is no consensus on the optimal protocol for management of subfertile women with adenomyosis. Modified downregulation could improve the clinical pregnancy rate by reducing the endometrial inflammatory reaction and/or myometrial contractility and their impact on uterine receptivity in women with moderate and severe adenomyosis of the uterus undergoing FTET. The MODA trial is designed to offer pragmatic, real-life evaluation of the optimal protocol for downregulation for this population during assisted conception treatments. Our findings will be published in peer-reviewed journals and presented at national and international scientific meetings and congresses. Ethical approval was granted by the NHS Research Ethics Committees (19/LO/1567).Trial registration numberNCT03946722.

INTRODUCTION AND OBJECTIVE:There is increased interest in using testicular sperm in assisted reproduction technology (ART) to improve outcomes after previous failed cycles. Mehta et al. (2015) repo...

Background: The first successful livebirth using warmed oocytes (vitrified by the GAVITM system) is reported in this paper. Embryologists throughout the world have vitrified oocytes using a manual technique which is susceptible to error and variation. In this era of automated laboratory procedures, vitrification was made semi-automatic by using the GAVITM system. Case Presentation: Donor oocytes were initially vitrified using the GAVITM system. They remained in the clinic’s oocyte bank until they were allocated to the patient. Donor oocytes were warmed as per Genea BIOMEDX protocol and inseminated to create embryos. Resulting embryos for the 42-year-old patient were cultured to the blastocyst stage, biopsied to perform PGT-A, using next generation sequencing and subsequently vitrified. The patient underwent a single euploid transfer in a frozen embryo transfer cycle which resulted in a healthy livebirth. Conclusion: The introduction of a semi-automated system should minimize the risk to the oocytes, standardize the procedure worldwide and potentially reduce the laboratory time taken by the embryologists. This case report demonstrates the safety of the technology used for vitrification, but larger randomized studies need to be performed to demonstrate the safety and efficacy of newer technologies like the GAVITM system before adopting it as a standard laboratory procedure.

During the first week of development, human embryos form a blastocyst comprised of an inner cell mass and trophectoderm (TE) cells, the latter of which are progenitors of placental trophoblast. Here we investigated the expression of transcripts in the human TE from early to late blastocyst stages. We identified enrichment of transcription factors GATA2, GATA3, TFAP2C and KLF5 and characterised their protein expression dynamics across TE development. By inducible overexpression and mRNA transfection we determined that these factors, together with MYC, are sufficient to establish induced trophoblast stem cells (iTSCs) from human embryonic stem cells. These iTSCs self-renew and recapitulate morphological characteristics, gene expression profiles and directed differentiation potential similar to existing human TSCs. Systematic omission of each or combinations of factors, revealed the critical importance of GATA2 and GATA3 for successful iTSC reprogramming. Altogether, these findings provide insights into the transcription factor network that may be operational in the human TE and broaden the methods for establishing cellular models of early human placental progenitor cells, which may be useful in the future to model placental-associated diseases.Summary statementTranscriptional analysis of human blastocysts reveals transcription factors sufficient to derive induced trophoblast stem cells from primed human embryonic stem cells.

Despite improvements in assisted reproduction techniques (ARTs), live birth rates remain suboptimal, particularly in women with advanced maternal age (AMA). The leading cause of poor reproductive outcomes demonstrated in women with AMA, as well as women with recurrent miscarriage and repetitive implantation failure, is thought to be due to high rates of embryonic aneuploidy. Preimplantation genetic testing for aneuploidies (PGT-A) aims to select an euploid embryo for transfer and therefore improve ART outcomes. Early PGT-A studies using fluorescent in situ hybridization on mainly cleavage-stage biopsies failed to show improved delivery rates and, in certain cases, were even found to be harmful. However, the development of comprehensive chromosome screening, as well as improvements in culture media and vitrification techniques, has resulted in an emerging body of evidence in favor of PGT-A, demonstrating higher implantation, pregnancy, and live birth rates. While there are concerns regarding the potential harm of invasive biopsy and the cost implications of PGT-A, the introduction of noninvasive techniques and the development of new high-throughput methods which lower costs are tackling these issues. This review aims to assess the evidence for PGT-A, address possible concerns regarding PGT-A, and also explore the future direction of this technology.

Study question What live birth rate do we see when we use testicular sperm in ART for non-azoospermic couples after at least one previous failed cycle? Summary answer In our cohort of couples 24% had a live birth using testicular sperm and therefore was not higher than national average ART rates. What is known already There is increased interest in using testicular sperm in assisted reproduction technology (ART) to improve outcomes after previous failed cycles. Mehta et al. reported results of a 50% live birth rate using testicular sperm in the first cycle for couples with oligospermia and a history of failed cycles with ejaculated sperm. We aim to audit our results in a similar population of couples. Study design, size, duration St Peters Andrology Centre in London, United Kingdom completed 128 surgical testicular sperm retrievals reviewed between the two-year period of 2018–2019. We conducted a retrospective audit of their paper-based records to identify those couples with injectable sperm on their semen analysis and who had previous cycles attempts using ejaculated sperm. Participants/materials, setting, methods We identified 27 couples who underwent testicular sperm extraction despite having an ejaculated semen analysis with injectable sperm and at least one previous failed cycle. A systematic review of their paper and electronic medical record was conducted to assess live birth rates and fertilization rates from ART. Main results and the role of chance Couples had an average male age of 41 (range 31–60) and an average female age of 38 (range 30–45). The men had an average serum testosterone of 15 nmol/L (range 8–35 nmol/L) and an average serum FSH of 8.9 IU/L (range 1.7–30 IU/L). 59% (n = 17) of men had a DNA fragmentation index completed with an average score of 41% (range 31%–51[Y1]%). In the women the mean serum anti-Müllerian hormone (AMH) was 15.8 pmol/l (range 1–64 pmol/l). With ejaculated sperm the fertilization rate was 59% (95% CI [27%, 59%]) and blastocyst conversion rate was 43% (95% CI [50%, 69%]). There was no statistical significance with testicular sperm where the fertilization rate was 58% (95% CI [51%, 65%]) and blastocyst conversion rate was 54% (95% CI [40%, 67%]). Overall, there were 7 clinical pregnancies in this population of couples. Of these clinical pregnancies, 2 miscarried and 5 progressed to a live birth. This audit yielded a live birth rate per cycle of 15% and a live birth rate per couple of 24%. Limitations, reasons for caution Limitations of the study are low number of patients and absence of a control group. Wider implications of the findings: We recommend caution and further analysis going forward using testicular sperm in ART where ejaculated sperm in available. Trial registration number Not applicable

Study question Does PGD treatment in couples with a history of RPL due to male translocations improve the outcome, increasing LBR and reducing miscarriage rate and time taken to live birth? Summary answer Live birth rate is significantly increased, miscarriage rate is significantly reduced using PGD. Time taken to achieve live birth rate is shorter in PGD treatment. What is known already Reciprocal translocation are the most common structural rearrangement in infertile men. The specific chromosomes and breakpoints involved might play an important role, often expressed as abnormal semen parameters or repeated pregnancy loss (RPL). The genetic counselling of these men remains challenging. Previous studies and meta-analysis performed showed no difference in live birth rate when comparing natural conception versus PGD treatment. However, the difference in miscarriage rate and time to live birth between PGD and natural conception has not been reported before in the medical literature. Study design, size, duration A systematic review of the literature was ­conducted through MEDLINE, EMBASE, and the Cochrane database up until December 2020. A comprehensive search yield 287 articles, 25 of which were included for abstract reading, finally, six were included in the meta-analysis. Participants/materials, setting, methods The six selected articles, reported on Live birth rate (LBR), miscarriage rate and time to live birth (TTLB) for natural conception compared to PGD for the same cohort of patients. All of the included articles were of retrospective design. The primary outcome was the comparison in LBR and the second outcome was the analysis in miscarriage rate and TTLB in the PGD group versus natural conception. Main results and the role of chance A total of 1438 couples that conceived naturally, had a LBR of 22.46%, compared with 43,17% among 681 couples that underwent PGD (0.53 95% CI (0.43-0.65) p o Regarding TTLB, the difference was not statistically significant, however it did reflect that PGD patients will have a shorter TTLB (3.56 95% CI (-0.88-8.00)p = 0.12). One of the studies included, took into account the waiting list to access PGD funding, prolonging therefore the TTLB in the PGD group. Limitations, reasons for caution The main limitation of this study is the low number of studies. TTLB should be interpreted with caution given that one of the articles included the time of the waiting lists. More studies could demonstrate a shorter time period for these couples to conceive and have a successful ongoing pregnancy. Wider implications of the findings First study to demonstrate the value of PGD in decreasing miscarriage rates in couples with RPL. Specially when counselling couples with history of RPL with male translocations. PGD should be offered in these couples to improve the outcome, and to diminish the physical, emotional and sequelae of RPL and TOP. Trial registration number not applicable

Study question Does a euploid embryo from one ovarian stimulation lead to the same live birth rate as a euploid embryo arising from multiple ovarian stimulations? Summary answer The live birth rate of a euploid embryo transferred is comparable irrespective of the number of ovarian stimulations required. What is known already Embryo transfer of a euploid embryo leads to a high live birth rate. Women with low ovarian reserve or poor responders may not have a euploid embryo from one cycle of ovarian stimulation and can be discouraged from undergoing preimplantation genetic testing for aneuploidy (PGT-A). Embryo batching from multiple cycles offers such patients a potential solution to increase their chance of achieving a euploid embryo. Study design, size, duration A retrospective analysis of 506 cycles of single euploid frozen embryo transfers (FET) from January 2015 to March 2020 was carried out. The indication for PGT-A was advanced maternal age, recurrent miscarriages or repetitive IVF failures. Only the first single euploid FETs per patient were included. Participants/materials, setting, methods Group A (N = 323) included women who had a normal ovarian reserve and only one cycle of ovarian stimulation before the FET, whilst Group B (N = 183) had low ovarian reserve or previous poor ovarian response requiring 2 or more cycles of ovarian stimulation. All embryos were biopsied at the blastocyst stage and subjected to a-CGH or NGS. Univariate statistical analysis with Chi-square or Wilcox Man U as required and multivariate logistic regression was performed with SPSS. Main results and the role of chance Group A and Group B were comparable in terms of BMI (average 22.3 vs 22.6), sperm origin, number of blastocysts biopsied (median N = 6, range 4–8), day 5 vs day 6 biopsy (day 5, 81.1% vs 75.4%, p = 0.130) and whether only one euploid embryo was available (42% vs 34%; p = 0.103). There was a significant difference in the number of eggs retrieved per cycle between the two groups (median 15 vs 9, p Multivariate logistic regression was performed to ascertain the effect of the treatment variables on the live birth rate. The number of oocyte collections was not a significant predictive factor (OR 1.19, 95% CI 0.72 - 1.96, p = 0.491), whilst an embryo biopsy performed on day 5 vs day 6, increased significantly the live birth rate (OR 2.58, 95% CI 1.61 - 4.13, p Limitations, reasons for caution The main limitation of this study is that it is retrospective, single centre and therefore vulnerable to confounding factors and bias. Wider implications of the findings: These results can be used to counsel and reassure women with poor response embarking on embryo batching and PGT-A that should a euploid embryo become available, their chance of success is unaffected by the number of cycles they undertake albeit the physical, emotional and financial burden of multiple ovarian stimulations Trial registration number Not applicable

Introduction To study whether paternal age exerts an effect, independent of maternal age, on the outcomes of fresh in vitro fertilization/ intracytoplasmic sperm injection (IVF/ICSI) cycles. Semen quality deteriorates with increasing paternal age; however, there is conflicting evidence for any impact paternal age may have on the outcome of IVF/ICSI. Several retrospective and prospective cohort studies have shown that paternal age increases the miscarriage rate and reduces the live birth rate. Some studies have shown no effect of paternal age on live birth rate or miscarriage rate. Studies involving donor oocytes have tended to show no independent effect of paternal age on assisted reproductive technology (ART) outcomes. The age at which paternal age may exert a significant deleterious effect on outcome is not known and there is no limit to paternal age in IVF/ICSI treatment. Material and methods A single-center retrospective cohort study was carried out at the Centre for Reproductive and Genetic Health, London, UK. Included in the analysis were all couples with primary or secondary infertility undergoing IVF/ICSI cycles in which the male partner produced a fresh semen sample and the cycle proceeded to fresh embryo transfer. All cycles of IVF/ICSI that used donor oocytes-donor sperm, frozen sperm, cycles leading to embryo storage and cycles including preimplantation genetic testing (PGT-A/PGT-M)-were excluded from analysis. The primary outcome was live birth rate and secondary outcomes were clinical pregnancy rate and miscarriage rate. Multivariate logistic regression analysis with live birth as a dependent variable and maternal and paternal age class as independent variables was performed. Results During the study period there were 4833 cycles, involving 4271 men, eligible for analysis; 1974/4833 (40.8%, 95% confiene intervals [CI] 39.5-42.2%) cycles resulted in a live birth. A significantly lower proportion of men over 51 years met World Health Organization semen analysis criteria (56/133, [42.1%, 95% CI 34.1-50.6]) compared with men under 51 years of age (2530/4138 [61.1%, 95% CI 60.0-62.6]) (p = 0.001). Both maternal and paternal age were retained in the multivariate model and for all maternal age subgroups the probability of live birth decreased with paternal age over 50 years (odds ratio [OR] 0.674, 95% CI 0.482-0.943) (p = 0.021). Paternal age over 50 years was not an independent predictor of miscarriage (OR 0.678, 95% CI 0.369-1.250) (p = 0.214). Conclusions Paternal age over 50 significantly affects the chance of achieving a live birth following ART. Paternal age does not independently affect the risk of miscarriage following ART. There should be a public health message for men not to delay fatherhood.

INTRODUCTION Social egg freezing enhances reproductive autonomy by empowering women with the capacity to delay their childbearing years, while preserving the opportunity to maintain biological relation with subsequent offspring. However, age-related obstetric complications, economic implications and the risk of unsuccessful future treatment make it a controversial option. Despite the upward trend in women electively cryopreserving their eggs, there is limited data about the women's perceptions, having undergone the process. The aim of this study was to investigate the motivations of women who have undergone social egg freezing, identify their perceptions following treatment, and assess potential feelings of regret. MATERIAL AND METHODS This cross-sectional survey, based at a fertility clinic in the UK, used an electronic questionnaire to assess the motivations and perceptions of women who underwent social egg freezing between 1 January 2008 and 31 December 2018. RESULTS One hundred questionnaires were distributed, and 85 women responded (85% response rate). The most frequent reason for freezing oocytes was not having a partner with 56 (70%) women saying it "definitely" influenced their decision. The majority of women (83%; n = 68) knew there was a chance of treatment failure in the future and that a live birth could not be guaranteed. More than half (n = 39; 51%) disagreed or strongly disagreed that the 10-year UK storage limit is fair. One-third of respondents (n = 17; 33%) felt the storage time should be indefinite and 29% (n = 15) believed it should be up to the age of 50. Twenty percent (n = 15) of the women who underwent social egg freezing have successfully had a baby or are currently pregnant, half (n = 8; 53%) of whom conceived spontaneously and a quarter (n = 4; 26%) used their stored oocytes. In all, 91% (n = 73) had no regrets over their decision to undergo social egg freezing. CONCLUSIONS We demonstrate herein important and novel insights into the motivations and perceptions of women from a UK population who have undergone social egg freezing. Despite potential physical, psychological, and financial burdens, only a small minority of women experience regret after social egg freezing. We also highlight clear discontent with the Human Fertilisation & Embryology Authority storage limit among social egg freezers in the UK.

The aim of this study was to compare clinical and laboratory outcomes between GnRHa, dual and HCG triggers in altruistic oocyte donation cycles. Normal or high responders were given either gonadotropin releasing hormone agonist (GnRHa) or a dual trigger of GnRHa and a low dose of human chorionic gonadotropin (HCG). Low responders were given HCG trigger. In 333 cycles, 232 (69.7%) received GnRHa trigger, 59 (17.7%) received dual trigger and 42 (12.6%) had HCG trigger. The total number of mature oocytes retrieved and cryopreserved were significantly higher in the GnRHa and dual trigger groups, compared to the HCG group (

Background: Within the ovary, the optimal growth of the follicle, oocyte maturation and ovulation are highly conditioned by the two-way cross talk and interactions between the oocyte and the immediate somatic cells, known as cumulus cells (CCs). This biological communication between cell lines triggered the interest in the study of CCs as a biomarker of oocyte competence. Case Presentation: The findings of a 45,X mosaic pattern on CCs from a female patient with unremarkable medical history are reported in this study. The patient came to the Centre for Reproductive and Genetic Health, London on 14th August 2019 for her first visit and the follow up procedures were done for her to determine underlying genetic status. For this purpose, four sources of DNA including CCs, blood lymphocytes, buccal cells and immature oocytes were analyzed in the present report. Conclusion: In the present case study, the hypothesis of the female patient being mosaic 45,X was confirmed although the degree of mosaicism and whether this was affecting the germinal line could not be determined. In the event of the discovery of a cell line with an apparently abnormal genetic makeup, genetic counselling is important in order to understand the implications from somatic to germinal cells for patients exploring fertility journeys

Introduction The effect of embryo quality on clinical outcomes of assisted reproductive technology following a double transfer is not well defined, with some studies suggesting that a low-quality embryo transferred with a high-quality embryo decreases the live birth rate (LBR), compared with transferring a single high-quality embryo. Our study examined whether the quality of a second blastocyst transferred affects the outcome, controlling for the number of the available high-quality blastocysts (HQB). Material and methods A historical cohort study of 2346 fresh blastocyst transfers in a single fertility clinic between 2013 and 2019. The main outcomes were pregnancy, miscarriage, live birth, and multiple gestation rates. Outcomes were compared between single embryo transfers with a high-quality blastocyst (SET-H), double embryo transfers with two HQBs (DET-HH), and transfers with one high-quality and one low-quality blastocyst (DET-HL). Outcomes were also assessed between SET and DET when only low-quality blastocysts were available. Results With one HQB available, DET-HL increased LBR (adjusted odds ratio [OR] 1.65, 95% CI 1.09-2.49) compared with SET-H, but increased multiple gestation rate (aOR 23.1, 95% CI 3.0-177.6). With two HQBs available, DET-HH was associated with a higher LBR (aOR 1.62, 95% CI 1.28-2.04) and lower miscarriage rate (aOR 0.56, 95% CI 0.40-0.80), but very high twin rate (aOR 49.8, 95% CI 24.3-102.1) compared with SET-H. A SET-H with at least one or more HQB available to freeze, compared with a SET-H with no other HQB available, had a higher LBR (aOR 1.69, 95% CI 1.17-2.45). When there were no HQBs available, compared with SET-L, a DET-LL had a higher live birth (aOR 3.20, 95% CI 1.78-7.703) and twin rate (aOR 3.72 × 1010 ) and a lower miscarriage rate (aOR 0.24, 95% CI 0.10-0.58). Conclusions When there is one HQB available, transferring an additional low-quality blastocyst would only slightly increase the LBR, but significantly increase the twin rate, therefore SET should be recommended. When two or more HQBs are available, SET-H would have a reasonably good chance of success without the very high twin rate associated with DET-HH. DET-LL when compared with SET-L, would increase the LBR, but increase the risk of multiple gestation.

Pregnancies at an advanced reproductive age are increasingly common. However, the safety of pregnancy remains a concern as maternal age is a recognized independent factor for various obstetric complications. Also, age is a risk factor for most systematic health problems and older women are more likely to enter into pregnancy with pre-existing conditions. At the moment there is no separate, structured guidance on preconception tests at advanced maternal age. However, the preconceptual period offers an ideal window to recognize and address underlying health issues, social issues and harmful lifestyle behaviours in order to optimize maternal health ultimately reducing infertility, perinatal morbidity and mortality. Preconception tests should be clinically relevant aiming to identify risk factors and address them to predict and prevent infertility and pregnancy complications. The importance of preconception care is magnified for women of advanced age for whom the risks are higher and the potential benefits greater.

Objective: To assess the relationship between number of oocytes retrieved during social egg freezing (SEF) cycles with various clinical, biochemical and radiological markers; e.g. age, body mass index (BMI), baseline anti-Mullerian hormone (AMH), antral follicle count (AFC), Oestradiol level (E2) and total number of follicles ≥12mm at trigger. Main outcome measures: To describe the characteristics and outcomes of women who underwent SEF. Methods: A retrospective cohort of women embarking on SEF between 2008 and 2018 from a single London UK fertility clinic. Results: 483 stimulation cycles were undertaken in 373 women. The median age at freeze was 38 (26-47) years. The median numbers of oocytes retrieved per cycle was 8 (0-37), and total oocytes cryopreserved 8 (0-45) per woman. BMI, E2 level and number of follicles ≥12mm at trigger were all significant predictors of oocyte yield. Multivariate analysis confirmed no significant relationship between AFC or AMH, whilst on univariate analysis statistical significance was proven. 36 women returned to use their oocytes, with 41 autologous egg thaw cycles undertaken. 12 successful livebirths were achieved by 11 women. The overall livebirth rate was 26.8% per cycle. No livebirths occurred in women ≥40 years old and 82% of all livebirths were in women aged 36-39 at freeze. Conclusions: This study demonstrates clinical, biochemical and radiological markers can predict oocyte yield in SEF cycles. However, subsequent reproductive outcomes highlight women embarking upon SEF should be encouraged to do so before the age of 37 years, and no later than 40 to optimise successful livebirth.

Background: The advent of ovarian stimulation within an in vitro fertilization (IVF) cycle has resulted in modifying the physiology of stimulated cycles and has helped optimize pregnancy outcomes. In this regard, the importance of progesterone (P4) elevation at time of human chorionic gonadotrophin (hCG) administration within an IVF cycle has been studied over several decades. Our study aimed to evaluate the association of P4 levels at time of hCG trigger with live birth rate (LBR), clinical pregnancy rate (CPR) and miscarriage rate (MR) in fresh IVF or IVF-ICSI cycles. Methods: This was a retrospective cohort study (n=170) involving patients attending the Centre for Reproductive and Genetic Health (CRGH) in London. The study cohort consisted of women undergoing controlled ovarian stimulation using GnRH antagonist or GnRH agonist protocols. Univariate and multiple logistic regression ana-lyses were used to evaluate the association of clinical outcomes. Differences were considered statistically significant if p£0.05. Results: As serum progesterone increased, a decrease in LBR was observed. Following multivariate logistical analyses, LBR significantly decreased with P4 thresholds of 4.0 ng/ml (OR 0.42, 95% CI:0.17-1.0) and 4.5 ng/ml (OR 0.35, 95% CI:0.12-0.96). Conclusion: P4 levels are important in specific groups and the findings were statistically significant with a P4 threshold value between 4.0-4.5 ng/ml. Therefore, it seems logical to selectively measure serum P4 levels for patients who have ovarian dysfunction or an ovulatory cycles and accordingly prepare the individualized management packages for such patients.

Objective To study the current evidence on the role of immunotherapy in IVF and in the management of recurrent pregnancy loss (RPL). Design Systematic review and meta-analysis. Setting A literature search was performed using MEDLINE, PUBMED, CINAHL, and EMBASE until May 2017. Only randomized controlled trials were included, and a meta-analysis was carried out where appropriate. Patient(s) Women undergoing IVF treatment with or without a history of recurrent implantation failure and women with idiopathic RPL. Intervention(s) Assessment of the efficacy of commonly used immunomodulators such as IV use of [1] immunoglobulin, [2] lymphocyte immunotherapy and [3] intralipid; intrauterine infusion of [4] granulocyte colony-stimulating factor and [5] peripheral blood mononuclear cells; subcutaneous administration of [6] TNF-alpha inhibitors, [7] leukaemia inhibitory factor; and oral administration of [8] glucocorticoids. Main Outcome Measure(s) The primary outcomes were live birth rate and miscarriage rate; secondary outcome was clinical pregnancy rate. Result(s) Of the 7,226 publications identified, 53 were selected during the initial screening; 30 satisfied the selection criteria and were included in this review. Conclusion(s) The available medical literature shows controversial results about the role of immunotherapy when used for improving reproductive outcomes. This study did not show a role for immunotherapy in improving the live birth rate in women undergoing IVF treatment or in the prevention of idiopathic RPL. Currently, immunotherapy should be used in the context of research and should not be used in routine clinical practice to improve reproductive outcomes.

Given the age-related decline in ovarian reserve and oocyte quality, it is unsurprising the global trend of deferring motherhood has resulted in increased levels of involuntary childlessness. The development of oocyte vitrification, with pregnancy and livebirth rates now comparable to using fresh oocytes, has provided an opportunity to cryopreserve oocytes electively for future use, empowering women with the capacity to delay their childbearing years. While it enhances reproductive autonomy, age-related obstetric complications, economic implications and the risk of unsuccessful future treatment make this a controversial therapeutic option. However, some women have no reasonable alternative, such as single women approaching their late thirties, in whom egg freezing, although not a guarantee against involuntary childlessness, offers hope by extending the window of opportunity to find a partner. Given the upward trend in women electively cryopreserving their eggs, it would appear that a new ice age, from a fertility perspective, is upon us.

Objectives: To investigate whether live birth (LB) is associated with oocyte yield and number of biopsied and suitable blastocyst to transfer following preimplantation genetic testing (PGT) for monogenic disorders (PGT-M) and chromosomal structural rearrangements (PGT-SR). Study design: All couples underwent controlled ovarian stimulation, blastocyst biopsy, vitrification and transfer of suitable embryo(s) in a frozen embryo transfer (FET) cycle. Results: Of 175 couples who underwent PGT treatment, 249 oocytes retrievals were carried out and 230 FET were subsequently undertaken. 122/230 (53%, 95% CI 47–59) FET resulted in a LB and 16/230 (7%, 95% CI 4–11) have resulted in ongoing pregnancies. 21/230 (9%, 95% CI 6–14) FET resulted in miscarriage and 69/230 (30%, 95% CI 24–36) concluded with failed implantation. Two (1%, 95% CI 0–3) transfers underwent termination for congenital malformation, with no evidence of misdiagnosis by prenatal testing. The relationship between number of oocytes retrieved and number of blastocysts biopsied and suitable embryos to transfer were significant (p = 0.00; Incidence rate ratio (IRR) 1.05; 95% 1.04–1.06; p = 0.00; IRR 1.04; 95%, 1.03–1.06), respectively. The number of oocytes collected (p = 0.007; OR 1.06; 95% CI 1.01–1.10), the number of blastocysts biopsied (p = 0.001; OR 1.14; 95% 95% CI 1.06–1.23) and the number of suitable embryos to transfer (p = 0.00; OR 1.38; 95% CI 1.17–1.64) were all significantly associated with the odds of achieving a LB. There is a 14% and 38% increased chance of a LB per additional blastocyst biopsied and suitable embryo to transfer, respectively. Conclusions: PGT-M and PGT-SR outcomes are significantly associated with egg yield, number of blastocysts to biopsy and suitable embryos to transfer. Keywords: Live birth, Preimplantation genetic testing, Monogenic disorders, Chromosomal rearrangements, Oocyte, Blastocysts

Introduction Intracytoplasmic morphologically selected sperm injection (IMSI) is one of the sperm selection techniques used for assisted reproduction which has been applied for a variety of indications including previously failed fertilization with intracytoplasmic sperm injection (ICSI). A Cochrane review1 found no difference in outcomes between either modality of sperm selection. Since the Cochrane review was published there have been a further two randomized controlled trials comparing IMSI and ICSI. This systematic review and meta-analysis aims to compare IMSI with ICSI as insemination methods regarding live birth rate and miscarriage rate. Material and methods Systematic review of randomized controlled trials, observational studies and similar reviews in electronic databases published before January 2018. Results We found nine randomized controlled trials, evaluating 1610 cycles of in vitro fertilization and 15 observational studies evaluating 1243 cycles of in vitro fertilization. Meta-analysis of the included randomized controlled trials showed no difference in the live birth rate or miscarriage rate between the ICSI and IMSI groups. Meta-analysis of five observational studies showed a significantly higher number of live births in the IMSI group than ICSI group (live birth rate odds ratio 1.47, 95% confidence interval 1.16-4.07), with a moderate degree of heterogeneity (I2 = 41%). Additionally, from six observational studies, a significantly lower miscarriage rate was observed in the IMSI group than in the ICSI group (odds ratio 0.51, 95% confidence interval 0.37-0.70, I2 = 0%). Conclusions Meta-analysis of randomized studies comparing IMSI to ICSI has not shown any difference in live birth rate and miscarriage rate. Meta-analysis of observational studies, which must be interpreted with caution, revealed an increased live birth rate and decreased miscarriage rate with IMSI vs ICSI.

PURPOSE: Does blastocyst morphology following euploid elective single embryo transfer (eSET) after preimplantation genetic testing for aneuploidies (PGT-A) via next generation sequencing impact clinical outcome? METHODS: Two hundred ninety-six patients underwent PGT-A. Of 1549 blastocysts, 1410 blastocysts had a conclusive result after PGT-A and were included for analysis. An eSET policy was followed in a frozen embryo replacement cycle. A total of 179 euploid blastocysts were thawed and transferred. Clinical outcomes were categorized in four different embryo quality groups: excellent, good, average and poor. RESULTS: Euploidy rate was 19/36 (52.7%, 95% CI 37–68), 199/470 (42.3%, 95% CI 38–47), 156/676 (23.0%, 95% CI 20–26) and 39/228 (17.1%, 95% CI 13–23) in the excellent, good, average and poor quality blastocyst groups, respectively. Fitted logistic regression analysis taking into account the following covariables: female, age, embryo chromosomal status and day of blastocyst development/biopsy showed that morphology was predictive of the comprehensive chromosome screening result (p < 0.05). A logistic regression analysis was also performed on clinical outcomes taking into account the effect of blastocyst morphology and day of blastocyst development/biopsy. None of the parameters were shown to be significant, suggesting morphology and day of blastocyst development/biopsy do not reduce the competence of euploid embryos (p > 0.05). CONCLUSIONS: After eSET, implantation rate was 80–86%; live birth rate per embryo transfer was 60–73% and clinical miscarriage rate was found to be < 10% and were not significantly affected by the embryo morphology. Results are concordant with those reported when using aCGH and highlights the competence of poor-quality euploid embryos.

Key content Preimplantation genetic diagnosis (PGD) was developed for couples at risk of transmitting a genetic disorder to their children. Before PGD, options available to these couples were remaining childless, prenatal diagnosis, gamete donation or adoption. Couples requesting PGD have in vitro fertilisation (IVF) to produce embryos for biopsy, even if they are fertile. Learning objectives To understand indications for PGD. To learn about the genetic work up required prior to undergoing IVF. To learn about IVF, embryogenesis, the biopsy process and when to transfer disease-free/carrier embryo(s). Ethical issues PGD for ‘saviour sibling’: should couples be offered human leucocyte antigen (HLA) matched treatment for a child who is in remission or who may develop the late onset condition? PGD for sex selection: should we offer sex selection for ‘family balancing’?

Purpose To compare in vitro fertilization (IVF) with intracytoplasmic sperm injection (ICSI) in regard to post-fertilization development and outcome with the purpose of ascertaining the most effective fertilization method for assisted reproduction. Methods A retrospective cohort study of 136 split IVF/ICSI cycles (where sibling oocytes are fertilized by two different methods using the same sperm sample). Results IVF-derived embryos developed to the blastocyst stage at a significantly faster rate than ICSI-derived embryos. There was no significant difference in fertilization or livebirth rates between the two fertilization methods. Conclusions For patients with sperm progressive motility ≥ 1.0 × 106/ml (who usually constitute the majority of patients), no significant difference between the two fertilization methods was found in regard to fertilization rate or livebirth rate. Remaining factors influencing choice between the two methods appear to be restricted to convenience, financial considerations and concern with regard to possible perpetuation of genetically linked infertility to future generations.

PURPOSE: The aim of the study is to investigate how blastocyst contraction behaviour affects the reproductive competence in high-quality euploid embryos. METHODS: Eight hundred ninety-six high-quality blastocysts derived from 190 patients (mean age 38.05 (SD = 2.9) years) who underwent preimplantation genetic testing for aneuploidies (PGT-A) from January 2016 to October 2017 were included in this study. PGT-A results were reported as euploid or aneuploid. Aneuploid embryos were sub-classified into three categories: monosomy, trisomy and complex aneuploid. Retrospective studies of time-lapse monitoring (TLM) of those embryos were analysed and reproductive outcome of transferred embryos was collected. RESULTS: A total of 234/896 were euploid (26.1%) whilst 662/896 (73.9%) blastocysts were proven to be aneuploid from which 116 (17.6%) presented monosomies, 136 (20.5%) trisomies and 410 (61.9%) were complex aneuploid. The most frequent chromosomal complements were trisomies affecting chromosome 21 and monosomies involving chromosomes 16 and 22. Data analysis showed a statistical difference in the number of contractions being reported greater in aneuploid when compared to euploid embryos (0.6 vs 1.57; p

This study aimed to determine whether karyomapping can be applied to couples requiring preimplantation genetic diagnosis (PGD) for single gene disorder (SGD) and/or chromosomal rearrangement. 75/82 (91.5%) and 6/82 (7.3%) couples were referred for autosomal SGD and X-linked disease, respectively. One couple (1.2%) was referred for SGD and chromosomal rearrangement. Of 608 embryos, 146 (24%, 95% CI 21-28) day-3 and 462 (76%, 95% CI 72-79) blastocyst biopsies were performed. A total of 81 embryo transfers were performed; 16/81 (20%) were following day-3 embryo biopsy, 65/81 (80%) were following blastocyst biopsy and cryopreserved embryo transfer. Of 81 embryo transfers with known pregnancy outcome, 51 (63%, 95% CI 52-73) were on-going pregnancies, 6/81 (7%, 95% CI 3-15) resulted in first trimester miscarriages and 24/81 (30%, 95% CI 21-40) were failed implantations. Of the 51 on-going pregnancies, 15 (29%, 95% CI 19-43) couples had a singleton live birth at the time of write up. There have been no reports of abnormal prenatal, genetic testing or diagnosis of phenotype at birth. Karyomapping is reliable, efficient and accurate for couples requiring PGD for SGD and/or chromosomal rearrangement. Additionally, it provides aneuploidy screening, minimising risks of miscarriage and implantation failure.

Objective To assess the survival rate of vitrified oocytes used in an egg recipient programme and compare the clinical outcomes of pregnancy and live-birth rates per warmed oocyte with fresh autologous oocytes. The differences in the obstetrical outcomes between the two groups were also studied. Design A prospective case control study from a single in-vitro fertilisaton (IVF) Centre in UK Setting Centre of Reproductive and Genetic Health (CRGH), London Population Vitrified oocytes from egg donors and autologous fresh oocytes from patients attending for an IVF cycle Methods The study group consisted of 1490 vitrified oocytes, which were obtained from 145 egg donors who underwent a stimulation cycle at CRGH Centre. The control group included 145 age-matched women who underwent intra cytoplasmic sperm injection (ICSI) treatment with their own oocytes (n = 1528). The clinical outcomes clinical pregnancy rates (CPR) and live-birth rates (LBR) and obstetrical outcomes (gestational age and weight at delivery) were compared between the two groups. Statistical analysis of the summary data and logistic regression analysis was performed using statistical packages (SPSS Version 23 and Stata 2015). The percentages of all parameters in the cases and control groups were compared by Fisher’s exact test. A statistical significance level of 5% was adopted throughout the study. Main outcome measures Survival rate per thawed oocyte, clinical pregnancy rate and live-birth rate per embryo transfer was compared to the autologous oocyte group Results The survival rate of vitrified oocytes was 73.6% (95% CI: 71.3–75.8%). The clinical pregnancy rate (per embryo transfer) using vitrified oocytes was found to be 51.8% compared to 59.3% in the control group. The live birth rate per embryo transfer in the vitrified oocyte group was 46% (95% CI 37.4–54.7%) compared to 57.1% (95% CI 48.5–68.5%) in the control group. The live-birth rate per thawed oocyte was found to be 4.2%. The gestational ages of the fetus at delivery in both the groups were comparable 39.0 (95% CI 32.7–41.9%) and 39.1 (95% CI 25.6–42.0) (p = 0.38). There was no statistically significant difference in the birth weight between the study and the control group 3100 g (750–4337) and 3232 g (1616–4500) respectively (p = 0.28). Conclusions This is the first study reporting on the efficacy of a vitrified donor oocyte programme from within the UK. There were no significant differences in the obstetrical outcomes between vitrified donor oocytes and autologous oocytes. The above data will be encouraging for women who are undertaking egg freezing for medical and or social reasons.

Introduction In the context of IVF, it has become imperative to emphasize on embryo evaluation in order to maximise assisted conception outcome while minimising the risk of multiple pregnancy. Despite the exponential rise in knowledge and understanding of the dynamic process of embryo development in the laboratory, the classical evaluation methods based on morphology are still considered as the gold standard methods to classify and select embryos. Morphology assessment helps embryologists to detect dysmorphic or arrested embryos but chromosomal abnormalities can still be only identified by preimplantation genetic testing for aneuploidies (PGT-A). It remains controversial the strength of morphology as a tool to ensure replacement of euploid blastocysts. The present research aims to corroborate whether blastocyst morphology, using a modified Gardner and Cornell's group scoring system, correlates with ploidy status of embryos analysed with next generation sequencing (NGS). We also wanted to ascertain whether biopsy of poor quality embryo yield a euploid embryo to transfer and thus result in a favourable outcome. This is the first data set describing the correlation between blastocyst morphology and embryo ploidy status using NGS and clinical outcomes. Materials and Methods 296 patients underwent PGT-A. Of 1,549 blastocysts, 1,410 blastocysts had a conclusive result after PGT-A and were included for analysis. An elective single embryo transfer (eSET) policy was followed in a frozen embryo replacement cycle. A total of 179 euploid blastocysts were thawed and transferred. Clinical outcomes were categorised in four different embryo quality groups: excellent, good, average and poor. Results Euploidy rates were 19/36 (52.7%, 95% CI 37-68), 199/470 (42.3%, 95% 38-47), 156/676 (23.0%, 95% CI 20-26) and 39/228 (17.1%, 95% CI 13-23) in the excellent, good, average and poor quality blastocyst groups, respectively. Fitted logistic regression analysis taking into account the following co-variables; female age, embryo chromosomal status, and day of blastocyst development/biopsy showed that good (OR 0.6; 95% CI 0.4-0.8;p=0.001), average (OR 0.5; 95% CI 0.4-0.7; p=0.001) and poor (OR 0.2;95% CI 0.1-0.5; p=0.005) morphology with excellent morphology being the reference group was predictive of the comprehensive chromosome testing result. A logistic regression analysis was also performed on clinical outcomes taking into account for the effect of blastocyst morphology and day of blastocyst development/biopsy. None of the parameters were shown to be significant suggesting morphology irrespective of the embryo quality category and day of blastocyst development/biopsy do not reduce the competence of euploid embryos (p>0.05). Conclusions After euploid eSET, implantation rate was 80-86%; live birth rate per embryo transfer was 60-73% and clinical miscarriage rate was found to be less than 10% and were not significanlty affected by the embryo morphology category. These results are concordant with those reported when using array compartive genomic hybridization and highlights the competence of poor quality euploid embryos.

The aim of the study is to investigate the regulation of DNA repair genes by microRNAs (miRNAs). miRNAs are short non-coding RNAs that regulate transcriptional and post-transcriptional gene silencing. Several miRNAs that are expressed during preimplantation embryo development have been shown or are predicted to target genes that regulate cell cycle checkpoints and DNA repair in response to DNA damage.This study compares the expression level of 20 miRNAs and 9 target transcripts involved in DNA repair. The statistical significance of differential miRNA expression between oocytes and blastocysts was determined by t test analysis using the GraphPad Prism v6 software. The possible regulatory roles of miRNAs on their target messenger RNAs (mRNAs) were analysed using a Pearson correlation test.This study shows for the first time that several miRNAs are expressed in human oocytes and blastocysts that target key genes involved in DNA repair and cell cycle checkpoints. Blastocysts exhibited statistically significant lower expression levels for the majority of miRNAs compared to oocytes (p 0.05). Correlation analyses showed that there was both inverse and direct association between miRNAs and their target mRNAs.miRNAs target many mRNAs including ones involved in DNA repair mechanisms. This study suggests that miRNAs and their target mRNAs involved in DNA repair are expressed in preimplantation embryos. Similar to the miRNAs expressed in adult tissues, these miRNAs seem to have regulatory roles on their target DNA repair mRNAs during preimplantation embryo development.

The sperm chromatin structure assay (SCSA) has been proposed as a useful addition to the battery of tests routinely used to explore semen quality and hence to give an indication of the likelihood of a successful pregnancy. As usually performed at present, the assay yields two main sperm variables, the DNA fragmentation index (DFI) and the high DNA stainability (HDS). In the present study 275 patients undergoing 215 in vitro fertilization (IVF) and 215 intracytoplasmic sperm injection (ICSI) cycles were studied with the purpose of defining the clinical significance of HDS in IVF and ICSI cycles. Using the Spearman correlation test there were no significant statistical relationships between %HDS and fertilization rate, rate of embryo growth, blastocyst rate, implantation rate, or live birth rate. Rate of pregnancy loss showed a negative relationship significant at the 0.05 level which is unexplained. It is not known whether the normal practice of using processed sperm for fertilization plays any part in this lack of a negative effect of HDS level upon the stages of the cycle. A total of 16 patients with HDS levels >28% had an average live birth rate of 47.8% and an average pregnancy loss of 8.7%, which compared favourably with the group of patients as a whole.

This is a retrospective study aiming to assess telomere length in human embryos 4 days post fertilization and to determine whether it is correlated to chromosomal ploidy, embryo developmental rate and patient age. Embryos were donated from patients undergoing treatment in the assisted conception unit. Seven couples took part, generating 35 embryos consisting of 1130 cells. Quantitative fluorescent in-situ hybridization (FISH) measured the telomere length of every cell using a pan-telomeric probe. Conventional FISH on six chromosomes was used to assess aneuploidy in the same cells. Maternal and paternal age, referral reason, embryo developmental rate and type of chromosomal error were taken into account. Chromosomally abnormal cells were associated with shorter telomeres than normal cells for embryos that were developmentally slow. Cells produced by women of advanced maternal age and those with a history of repeated miscarriage tended to have substantially shorter telomeres. There was no significant difference in telomere length with respect to the rate of embryo development 5 days post fertilization. Telomeres play an important role in cell division and shorter telomeres may affect embryonic ploidy. Reduced telomere length was associated with aneuploid cells and embryos from women of advanced maternal age.

Germinal mosaicism in a parent is considered to be a rare cause of aneuploidy in the offspring. The aim of this study was to assess the incidence of pre-meiotic errors, indicative of germinal mosaicism, leading to aneuploidy compared with those that occur at meiosis I. The material consisted of 126 oocytes, unexposed to sperm, donated by 57 women with an average maternal age of 35. The oocytes were at various stages of maturity and were analysed by array comparative genomic hybridisation. Of these, 102 gave conclusive results, comprising 47 that were immature, at the germinal vesicle (GV) or metaphase I stage (MI); 34 complete metaphase II-first polar body (MII-PB) complexes together with 21 incomplete complexes. Oocytes at the GV or MI stage provide direct evidence of pre-meiotic aneuploidy. Complete MII-PB complexes with the expected reciprocal gains/losses provide information on MI errors; those with non-reciprocal gains have pre-meiotic errors. Overall, 29 oocytes were aneuploid, and the source of the error was known for 21. In 8 (from 7 women) the error was pre-meiotic consisting of 4 MI oocytes and 4 MII-PB complexes with non-reciprocal gains. The remaining 13 were the result of errors at meiosis I. Although pre-meiotic errors occurred in only 10% of informative oocytes, most notable was the fact that for those oocytes where the source of the error was known, 38% were caused by germinal mosaicism compared with 62% that were the outcome of a meiosis I error. None of the women with germinal mosaicism were infertile.

The aim was to investigate the influence of various biological factors upon the outcome of intrauterine insemination (IUI). The total IUI history (856 cycles) of 352 couples was studied. Live-birth showed a strong negative correlation with female age but no correlation with male age. Antimüllerian hormone (AMH) and antral follicle count (AFC) correlated negatively with female age, and follicle stimulating hormone (FSH) correlated positively. Significant thresholds were found for all three variables, and also for total motile count (TMC) in the prepared sperm. Calculating pregnancy losses per positive pregnancy showed a strong correlation with increasing female age. This was highly significant for biochemical losses but not for fetal heart miscarriages. Male age had no effect on rate of pregnancy loss. In conclusion, female age, FSH, AMH and TMC are good predictive factors for live-birth and therefore relate to essential in vivo steps in the reproductive process.

What's known on the subject? and What does the study add? The management of patients with non-obstructive azoospermia (NOA) and some cases of obstructive azoospermia involves testicular sperm extraction (TESE or micro-dissection TESE) combined with intracytoplasmic sperm injection (ICSI). Several studies have investigated the effect of the male age, the cause of azoospermia, testicular histopathology, the type of sperm used, and the use of pentoxyphilline, on the ICSI cycle outcome in men with azoospermia. The present study showed that none of these factors influenced the ICSI outcome in men with azoospermia, thus once sperm is found in an azoospermic male, no other male factor seems to influence the ICSI outcome. To our knowledge this is the first study to comment on the outcome of ICSI in men with NOA based on testicular histopathology. Objectives To access the effect of: male age, the cause of azoospermia (obstructive azoospermia vs non-obstructive azoospermia [NOA]), testicular histopathology, the type of sperm used (fresh vs frozen-thawed), and the use of pentoxyphilline on the intracytoplasmic sperm injection (ICSI) cycle outcome in men with azoospermia. To our knowledge this is the first study to comment on the outcome of ICSI in men with NOA based on testicular histopathology. Patients and Methods A retrospective analysis of 137 testicular sperm extraction-ICSI cycles performed between 2001–2010, involving 103 men with azoospermia, with 26 couples having repeat cycles. Results Analysis of the results did not show any statistically significant differences in the fertilization, embryo cleavage, clinical pregnancy, live birth and miscarriage rates in relation to the male age, cuase of azoospermia, testicular histopathology, type of sperm used and the use of pentoxyphilline. Conclusion Once sperm is found in a man with azoospermia, no other male factor seems to influence the ICSI outcome.

From 2006 to 2011, Roslin Cells Ltd derived 17 human embryonic stem cells (hESC) while developing (RCM1, RC-2 to -8, -10) and implementing (RC-9, -11 to -17) quality assured standards of operation in a facility operating in compliance with European Union (EU) directives and United Kingdom (UK) licensure for procurement, processing and storage of human cells as source material for clinical application, and targeted to comply with an EU Good Manufacturing Practice specification. Here we describe the evolution and specification of the facility, its operation and outputs, complementing hESC resource details communicated in Stem Cell Research Lab Resources.

Mosaicism in certain dominant disorders may result in a 'non-Mendelian' transmission for the causative mutation. Preimplantation genetic diagnosis (PGD) is available for patients with inherited disorders to achieve an unaffected pregnancy. We present our experience for two female patients with different dominantly inherited autosomal disorders; neurofibromatosis type 1 (NF1) and tuberous sclerosis complex type 2 (TSC2).PGD protocol development was carried out using single cells from the patients. PGD was carried out on polar bodies and different embryonic cells.Protocol development for NF1 using lymphocytes from the patient suggested mosaicism for the mutation. This was supported further by quantitative fluorescent-PCR performed on genomic DNA. During PGD, polar bodies and blastomeres lacked the mutation that probably was absent or present at very low levels in the patient's germline. Single lymphocyte analysis during protocol development for TSC2 did not indicate mosaicism; however, analysis of single buccal cells and multiple embryo biopsies across two consecutive IVF/PGD cycles confirmed gonosomal mosaicism.The trend in PGD is for blastocyst biopsy followed by whole genome amplification, eliminating single cell analysis. In the case of certain dominantly inherited disorders, pre-PGD single cell analysis is beneficial to identify potential mosaicism that ensures robust protocols. © 2016 John WileySons, Ltd.