Healthy Live Birth Research Articles

Synchronised approach for intrauterine insemination in subfertile couples.

In many countries intrauterine insemination (IUI) is the treatment of first choice for a subfertile couple when the infertility work up reveals an ovulatory cycle, at least one open Fallopian tube and sufficient spermatozoa. The final goal of this treatment is to achieve a pregnancy and deliver a healthy (singleton) live birth. The probability of conceiving with IUI depends on various factors including age of the couple, type of subfertility, ovarian stimulation and the timing of insemination. IUI should logically be performed around the moment of ovulation. Since spermatozoa and oocytes have only limited survival time correct timing of the insemination is essential. As it is not known which technique of timing for IUI results in the best treatment outcome, we compared different techniques for timing IUI and different time intervals. To evaluate the effectiveness of different synchronisation methods in natural and stimulated cycles for IUI in subfertile couples. We searched for all publications which described randomised controlled trials of the timing of IUI. We searched the Cochrane Menstrual Disorders and Subfertility Group Specialised Register, Cochrane Central Register of Controlled Trials (CENTRAL) (1966 to October 2014), EMBASE (1974 to October 2014), MEDLINE (1966 to October 2014) and PsycINFO (inception to October 2014) electronic databases and prospective trial registers. Furthermore, we checked the reference lists of all obtained studies and performed a handsearch of conference abstracts. Randomised controlled trials (RCTs) comparing different timing methods for IUI were included. The following interventions were evaluated: detection of luteinising hormone (LH) in urine or blood, single test; human chorionic gonadotropin (hCG) administration; combination of LH detection and hCG administration; basal body temperature chart; ultrasound detection of ovulation; gonadotropin-releasing hormone (GnRH) agonist administration; or other timing methods. Two review authors independently selected the trials, extracted the data and assessed study risk of bias. We performed statistical analyses in accordance with the guidelines for statistical analysis developed by The Cochrane Collaboration. The overall quality of the evidence was assessed using GRADE methods. Eighteen RCTs were included in the review, of which 14 were included in the meta-analyses (in total 2279 couples). The evidence was current to October 2013. The quality of the evidence was low or very low for most comparisons . The main limitations in the evidence were failure to describe study methods, serious imprecision and attrition bias.Ten RCTs compared different methods of timing for IUI. We found no evidence of a difference in live birth rates between hCG injection versus LH surge (odds ratio (OR) 1.0, 95% confidence interval (CI) 0.06 to 18, 1 RCT, 24 women, very low quality evidence), urinary hCG versus recombinant hCG (OR 1.17, 95% CI 0.68 to 2.03, 1 RCT, 284 women, low quality evidence) or hCG versus GnRH agonist (OR 1.04, 95% CI 0.42 to 2.6, 3 RCTS, 104 women, I(2) = 0%, low quality evidence).Two RCTs compared the optimum time interval from hCG injection to IUI, comparing different time frames that ranged from 24 hours to 48 hours. Only one of these studies reported live birth rates, and found no difference between the groups (OR 0.52, 95% CI 0.27 to 1.00, 1 RCT, 204 couples). One study compared early versus late hCG administration and one study compared different dosages of hCG, but neither reported the primary outcome of live birth.We found no evidence of a difference between any of the groups in rates of pregnancy or adverse events (multiple pregnancy, miscarriage, ovarian hyperstimulation syndrome (OHSS)). However, most of these data were very low quality. There is insufficient evidence to determine whether there is any difference in safety and effectiveness between different methods of synchronization of ovulation and insemination. More research is needed.

Blastocysts can be rebiopsied for preimplantation genetic diagnosis and screening

To evaluate the clinical value of re-examining the test-failure blastocysts in preimplantation genetic diagnosis/screening cycles. Retrospective study. University-affiliated center. Women with test-failure blastocysts cryopreserved in preimplantation genetic diagnosis/screening cycles. Cryopreserved test-failure blastocysts were warmed and underwent a second round of biopsy, single nucleotide polymorphism microarray analysis, and vitrification, and the normal blastocysts were warmed again for ET. The percentage of test-failure blastocysts for transfer, the implantation rate per transferred blastocyst, and the live birth rate. A total of 106 test-failure blastocysts from 77 cycles were warmed for re-examination. A total of 73 blastocysts that completely expanded were considered to have survived the warming process and were successfully rebiopsied. After single nucleotide polymorphism array analysis, 70 blastocysts yielded whole genome amplification product, and 31 had normal chromosomes (44.3%). A total of 19 normal blastocysts were warmed for ET, of which 18 survived and were transferred. The clinical pregnancy rate (implantation rate) was 50.0% in 10 single blastocyst transfer cycles, and all the implanted blastocysts resulted in healthy live births. Test-failure blastocysts that survived from the first warming procedure can tolerate a second round of biopsy, vitrification, and warming, have a high chance of having normal chromosomes, and are worth being re-examined.

Proteomic analysis of equine amniotic membrane: characterization of proteins.

Human amniotic membrane (AM) has been used as a biomaterial for surgical wound skin and ocular surface reconstruction for several years. Currently, equine AM has been used for corneal reconstruction in several animal species, and appears to have the same properties as human AM. Despite the observed positive healing abilities of this tissue in horses with ulcerative keratitis the proteins of equine AM have not been described. To identify proteins known to be associated with corneal healing from frozen equine AM. Placentas were acquired from healthy live foal births from a local Thoroughbred breeding farm. The amnion was removed from the chorion by blunt dissection, washed with phosphate-buffered saline (PBS), and treated with 0.05% trypsin and 0.02% ethylene diaminetetraacetic acid in PBS. Amnion was attached to nitrocellulose paper (epithelial side up), and cut into 4 × 4 cm pieces. The sheets were frozen at -80 °C. The protein samples were solubilized, and analyzed by 2D gel electrophoresis and shotgun proteomics. A reference identification map of the equine AM proteins was produced and 149 different proteins were identified. From gel-based proteomics, 49 spots were excised and 43 proteins identified by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Shotgun proteomics identified 116 proteins with an overlap of 10 proteins in both analyses. We have described a reference map for equine AM proteins that may provide a background to explain the positive results found in horses with ulcerative keratopathies using this biomaterial.

Introduction: Choosing the main outcome of an infertility trial is harder than you think

Clinical trials in infertility choose from a variety of outcomes including change in some surrogate marker of gamete quality to healthy live birth. Incomplete reporting of outcomes makes it difficult to compare studies and to determine the clinical impact of infertility treatments. In this Views and Reviews, we explore the merits of collecting various outcomes of interest in infertility trials from the vantage point of infertility specialists, an obstetrician, and a pediatrician. These articles support more complete reporting of maternal, paternal, fetal, and infant outcomes from infertility trials to improve patient care and ultimately public health.

An analysis of pregnancy outcome in dichorionic and monochorionic twins given special antenatal and intranatal care: a four-year survey.

The present study aims to analyze perinatal outcomes in twins given special care during pregnancy and labor and to compare fetal and neonatal outcomes in dichorionic twins with monochorionic twins. Eighty eight (88) twin pregnancies booked for care at a tertiary care Fetal Medicine centre were included in this study. The maternal demographic variables, course of pregnancy, fetal problems, and specialized fetal therapeutic and diagnostic interventions were noted. The above parameters were compared in the sub-groups of dichorionic and monochorionic twin pregnancies and related to the perinatal outcome. Statistical analysis was done using the student's t test and the two-tailed chi sqaure tests with Yate's continuity correction. A p value <0.05 was considered as significant. Mean maternal age was 30.34+4.81years (range 19-48). 81% of the twins were DCDA, and 19% were MCDA. The mean gestational age at delivery was 34.4±3.5weeks, and this was not significantly different in MCDA and DCDA groups. Serious fetal problems warranting intervention at the time of initial referral were significantly higher in MCDA twins although overall perinatal outcome in both groups were not different. Specialized care during pregnancy and labor including active fetal surveillance and therapeutic intervention when indicated improves the perinatal outcome in twin pregnancies and ensured at least one healthy live birth in over 90% cases, although with an increase in late prematurity.

Diagnóstico genético preimplantacional: alcances y límites

El diagnóstico genético preimplantacional (DGP) es el estudio de alteraciones cromosómicas y genéticas en el embrión, antes de su transferencia a la madre. Permite conocer aquellos embriones libres de anomalías cromosómicas o mutaciones genéticas. En esta revisión se describe la experiencia de un centro de reproducción asistida en Perú, realizando diagnóstico genético preimplantacional a embriones en día 3 de desarrollo. Se muestra resultados de cinco años, con 711 ciclos. El diagnóstico genético preimplantacional sería una herramienta eficaz para seleccionar los embriones con mayor potencial de implantación, que puedan dar origen a un nacido vivo sano.

Is it acceptable to destroy or include human embryos before day 5 in research programmes?

Day-3 poor-quality embryos (PQE) from IVF–embryo transfer cycles are usually destroyed or are included in research programmes. Knowing that these embryos have the ability to evolve to the blastocyst stage and yield embryonic stem cell lines, this study postulated that they could also give rise to live births. This is a prospective study including 186 IVF–embryo transfer candidates who had obtained at least one supernumerary PQE on day 3. PQE were kept for extended culture and high-quality blastocysts were frozen. A total of 620 PQE were eligible for the study, 217 (35.0%) reached the blastocyst stage and 73 (33.6%) were frozen. Blastulation rates were 7-fold higher (OR 7.29, 95% CI 5.01–10.61) in embryos compacted on day 4. Of the frozen blastocysts, 40 were thawed during 33 thawed blastocyst transfer cycles, which led to 10 clinical pregnancies. These pregnancies resulted in five miscarriages and five healthy live births at full term. PQE may achieve their development to the blastocyst stage, be frozen–thawed and harbour reasonable implantation potential. These results, thereby, raise an ethical issue regarding the fate reserved to PQE.In most IVF clinics around the world, day-3 embryos are submitted to embryo classification systems to select embryos with the higher implantation potential. This classification is usually based on the evaluation of the number of blastomeres, the degree of fragmentation, the symmetry of the cells and the presence of multinucleation. Thus, some embryos are described as poor-quality embryos (PQE) and may be destroyed or included in research programmes. Knowing that PQE have the ability to evolve to the blastocyst stage and yield embryonic stem cell lines, the authors hypothesized that PQE could also give rise to live births. They conducted a prospective study including 186 IVF–embryo transfer candidates who had obtained at least one supernumerary PQE on day 3. PQE were kept for extended culture and high quality blastocysts were frozen. Out of 620 PQE eligible for this study, 217 (35.0%) reached the blastocyst stage and 73 (33.6%) were frozen. Of these blastocysts, 40 were thawed during 33 thawed blastocyst transfer cycles, which led to 10 clinical pregnancies. These pregnancies resulted in five miscarriages and five healthy live births at full term. PQE may achieve their development to the blastocyst stage, be frozen–thawed and harbour a reasonable chance of implantation. These results, thereby, raise a ethical issue regarding the fate of PQE.

Healthy live birth using theophylline in a case of retrograde ejaculation and absolute asthenozoospermia

To analyze whether the use of ready-to-use theophylline is a feasible option in a case of retrograde ejaculation and absolute asthenozoospermia. Case report. In vitro fertilization unit of a public hospital. Thirty-one-year-old nulliparous woman, and 39-year-old male with retrograde ejaculation and absolute asthenozoospermia. Retrieval of postejaculatory urine, restoration of motility using a methylxanthine, intracytoplasmic sperm injection, single-embryo transfer. Sperm motility, fertilization, embryo quality, live birth. Successful fertilization and a single-embryo transfer resulted in a healthy live birth. Theophylline turned out to be a safe, efficient agent for stimulating immotile spermatozoa in patients with retrograde ejaculation.

Effects of immigrant status on Emergency Room (ER) utilisation by children under age one: a population-based study in the province of Reggio Emilia (Italy).

BackgroundThe primary aim of this study was to assess the effect of immigrant status on Emergency Room (ER) utilisation by children under age one, considering all, non-urgent, very urgent, and followed by hospitalisation visits. The second aim was to investigate the role played by mother’s educational level in the relationship between citizenship and ER utilisation.MethodsThe cohort study included all healthy singleton live births in the years 2008–2009 and residing in the province of Reggio Emilia, followed for the first year of life in order to study their ER visits. The outcomes were the ER utilisation rate for all, non-urgent, very urgent, and followed by hospitalisation visits. The main explanatory variable was mother’s citizenship. Other covariates were mother’s educational level, maternal age, parity, and child gender. Multivariate analyses (negative binomial regression and zero inflated when appropriate) were performed. Adjusted utilisation Rate Ratios (RR) and their 95% Confidence Intervals (95% CI) were calculated. Trend for age in months by citizenship is depicted.ResultsThere were 3,191 children (36.4%) with at least one ER visit in the first year of life. Adjusted RR show a significantly greater risk of ER visit for immigrants than for Italians: (RR 1.51; 95% CI 1.39-1.63). Immigrants also had a higher risk of non-urgent visits (RR 1.72; 95% CI 1.48-2.00) and for visits followed by hospitalizations (RR 1.58; 95% CI 1.33-1.89). For very urgent visits, the immigrants had a slightly higher risk compared to Italians (RR 1.25; 95% CI 0.98-1.59).The risk of ER visits is higher in the first two months of life (RR1stvs 3rd-12th 2.08; 95% CI 1.93-2.24 and RR 2ndvs 3rd-12th 1.45; 95% CI 1.33-1.58, respectively). Considering all visits, the ER utilisation rate was inversely related with maternal education only for Italians (low educational level 44.0 and high educational level 73.9 for 100 children; p value for trend test < 0.001).ConclusionsOur study observed a higher use of ER services by immigrant children and, to a lesser extent, by children of less educated Italian mothers. In immigrants, the excess is mostly due to non-urgent visits and only slightly to high acute conditions.

A novel method for human oocyte vitrification with a closed device using super-cooled air

Human oocytes have been successfully vitrified with open systems that involve direct contact with liquid nitrogen. Concerns have been raised regarding the safety and sterility of this methodology. It is therefore highly desirable to use a device that does not utilize direct contact with liquid nitrogen during vitrification and storage. It is thought, however, that the high cooling rate afforded by direct liquid nitrogen contact is essential for human oocyte vitrification. Human oocytes were collected from donors and vitrified/warmed for transfer into recipients. Following oocyte warming; fertilization, embryo development, implantation, biochemical pregnancy and clinical pregnancy rates were recorded. The results were compared retrospectively to data from non-vitrified oocytes within the same time period. Almost 600 oocytes from 53 donors were vitrified using a newly developed vitrification medium specifically for oocytes (RapidVit and RapidWarm Oocyte: Vitrolife AB) and a closed vitrification device that uses a straw with super-cooled air (Rapid-i™: Vitrolife AB). Oocytes were collected from 53 oocyte donors (mean age of 24.6 ± 4.0) and vitrified/warmed. The survival and fertilization rates were 94 % and 76 %, respectively. A mean number of 1.8 ± 0.5 embryos were transferred on day 5/6 per recipient (mean age 40.4 ± 4.6). The implantation rate was 34.4 % (33/96). The biochemical and clinical pregnancy rates were 65 % (33/51) and 51 % (26/51), respectively. The ongoing (>12 weeks) pregnancy rate was 49 % with 5 healthy live births reported so far. These results were not statistically different from outcomes with non-vitrified oocytes. This study demonstrates that, despite lower cooling rates, human oocytes can be vitrified without direct contact with liquid nitrogen in a closed device (Rapid-i™). A closed system that uses super-cooled air to vitrify removes the potential concerns regarding contamination from liquid nitrogen.

Pregnant rate and pregnancy-relating factors of patients with early endometrial carcinoma and severe atypical hyperplasia of endometrium after fertility-preserving treatment by progestin

To summarize the pregnant rate of patients with early endometrial carcinoma and severe atypical hyperplasia after fertility-preserving treatment and analyze their pregnancy-relating factors. Endometrial curettage was used to evaluate the therapy response of endometrium after every 3 months of administration of high-dose progestin as fertility-sparing treatment for 51 patients with stage I endometrial carcinoma or severe endometrial atypical hyperplasia from Jun. 1996 to Jan. 2010. Individualized maintained treatment was given to patients after achieving complete remission of the endometrium. Pregnant results and pregnancy-relating factors were analyzed retrospectively. The median age of all the 51 patients was 29 years old. Forty-five (88%, 45/51) achieved complete response. Of the 34 cases who desired to conceive after complete response, 16 of them had 22 pregnancies, the pregnant rate was 47% (16/34); and 12 women obtained healthy live birth baby, the fertility rate was 35% (12/34). The pregnant rate of patients at age >35 or ≤ 35 was 0/2 and 50% (16/32) respectively (P > 0.05). The pregnant rate of patients with or without infertility was 40% (8/20) and 8/14, with endometrial cancer or severe atypical hyperplasia was 40% (10/25) and 6/9, respectively (all P > 0.05). The pregnant rate of patients who received in vitro fertilization-embryo transfer, ovulation promotion, or no treatment was 7/7, 6/16 and 3/11 respectively (P < 0.01). Fertility-preserving treatment for early endometrial cancer and severe atypical hyperplasia with high-dose progestin could achieve higher response rate. Assisted reproductive technologies could significantly increase the chance of conception.

Female obesity: short- and long-term consequences on the offspring

The worldwide prevalence of obesity has risen over the past few decades and women are currently more likely than ever to enter pregnancy obese. Pre-pregnancy obesity and excessive gestational weight gain increase miscarriage rates and obstetric and neonatal complications, which result in a lower healthy live birth rate. In addition to its negative consequences for the mother, obesity has been shown to be an important risk factor for chronic illnesses, such as cardiovascular disease, metabolic syndrome and type 2 diabetes in the adolescence and adulthood of the offspring. Moreover, maternal obesity causes psychological problems, physical disabilities and higher healthcare costs. Fetal programming of metabolic function induced by obesity, through physiological and/or epigenetic mechanisms, may have an intergenerational effect and could, thus, perpetuate obesity in the next generation. In order to break this vicious circle and avoid serious short- and long-term negative outcomes for both mothers and fetuses, the prevention and adequate management of obesity and gestational weight gain are essential.

Modelling a risk classification of aneuploidy in human embryos using non-invasive morphokinetics

This study determined whether morphokinetic variables between aneuploid and euploid embryos differ as a potential aid to select euploid embryos for transfer. Following insemination, EmbryoScope time-lapse images from 98 blastocysts were collected and analysed blinded to ploidy. The morphokinetic variables were retrospectively compared with ploidy, which was determined following trophectoderm biopsy and analysis by array comparative genomic hybridization or single-nucleotide polymorphic array. Multiple aneuploid embryos were delayed at the initiation of compaction (tSC; median 85.1 hours post insemination (hpi); P=0.02) and the time to reach full blastocyst stage (tB; median 110.9hpi, P=0.01) compared with euploid embryos (tSC median 79.7 hpi, tB median 105.9 hpi). Embryos having single or multiple aneuploidy (median 103.4 hpi, P=0.004 and 101.9 hpi, P=0.006, respectively) had delayed initiation of blastulation compared with euploid embryos (median 95.1hpi). No significant differences were observed in first or second cell-cycle length, synchrony of the second or third cell cycles, duration of blastulation, multinucleation at the 2-cell stage and irregular division patterns between euploid and aneuploid embryos. This non-invasive model for ploidy classification may be used to avoid selecting embryos with high risk of aneuploidy while selecting those with reduced risk.Chromosome anomalies (aneuploidy) are prevalent in human embryos and result in either IVF failure, miscarriage or the birth of children with chromosome disorders. After IVF, it is imperative to try to detect aneuploid embryos in order to transfer an embryo with a normal chromosome copy number (euploid) to maximize the chances of a successful and healthy live birth. Currently, the only option is to biopsy a cell (or cells) from the embryo and analyse these using complex and expensive genetic technology. Novel uninterrupted culturing methods using the EmbryoScope now enables detailed observation of the each embryo via time-lapse photography during the whole culture period, for up to 6days. This new technology has permitted us to observe the development of human embryos in detail after IVF, providing analysis of complex patterns of cell division and cell movement (morphokinetics). As far as is known for the first time, we have demonstrated differences between aneuploid and euploid embryos based on their morphokinetic patterns, whereby aneuploid embryos are significantly delayed in reaching the later stages of development, during days 4 and 5 in culture. We postulate that, by using the unique, non-invasive and specifically designed models or algorithms developed in this study, embryologists can make more informed choices on the most viable embryo to select for transfer and reduce the risk of selecting an aneuploid embryo.

Restoring fertility after ovarian tissue cryopreservation: a half century of research

Tissue transplantation and in vitro ovarian follicle culture have been investigated as alternative techniques to restore fertility in young women who are facing fertility-threatening diseases or treatments following ovarian tissue cryopreservation. Although transplants of fresh or frozen ovarian tissue have successfully yielded healthy live births in different species including humans, the risks of reintroducing cancer cells back into the patient, post treatment, have limited its clinical purpose. The in vitro ovarian follicle culture minimizes these risks and provides a way to harvest more mature oocytes, however its clinical translation has yet to be determined. Not only is it possible for tissue cryopreservation to safeguard fertility in cancer patients, this technique also allows the maintenance of germplasm banks for animals of high commercial value or for those animals that are at risk of extinction. Given the importance of managing female genetic material, this paper reviews the progress of the methods used to preserve and restore female fertility in different species to demonstrate the results obtained in the past 50 years of research, the current achievements and the future directions on this field.

Mission possible: RNA interference rescues the hERG current

The congenital long QT syndrome (LQTS) is an inherited disease that causes cardiac arrhythmias and sudden cardiac death in the absence of evident structural heart defects. It is mainly attributed to genetic mutations that affect the function of ion channels that regulate the action potential waveform. LQTS is characterized by an abnormal prolongation of cardiac repolarization, leading to an increased QT interval, notched or biphasic T waves, and T-wave alternans. Affected patients are at a higher risk for developing arrhythmias, including polymorphic ventricular tachycardia, torsades de pointes, and ventricular fibrillation. The estimated prevalence of LQTS (based on electrocardiographic-guided identification of disease-causing mutations among whites) is approximately 1:2500 in apparently healthy live births. LQTS was initially described in the Jervell and Lange-Nielsen syndrome, where QT prolongation and deafness were present. A few years later, an autosomal dominant form of LQTS, in the absence of deafness, was reported. During the last decade, 13 LQTS variants have been described. Mutations in KCNQ1, KCNH2, and SCN5A genes account for approximately 90% of the positivegenotype cases of LQTS and underlie LQTS1, LQTS2, and LQTS3. Those genes encode Kv7.1, Kv11.1, and Nav1.5, respectively. Genetic analyses have identified more than 200 LQTS2causing mutations in KCNH2. Missense mutations leading to single amino acid substitutions in hERG (human ethera-gogo-related gene) protein—the alpha subunit of the rapid delayed rectifier current (IKr)—are the predominant abnormality. At the functional level, these mutations result in a decreased IKr. 11 It has been shown that LQTS2 mutations reduce IKr through a variety of mechanisms including aberrant hERG protein folding, trafficking, or ion-channel gating. Although not all identified mutations have been investigated, the majority of the mutations produce failure of the hERG subunits to exit the endoplasmic

Comprehensive chromosome screening (CCS) with vitrification results in improved clinical outcome in women >35 years: a randomized control trial

OBJECTIVE: Chromosome aneuploidy accounts for>70% of first trimester pregnancy loss, with maternal aging being the most significant risk factor. If demonstrated to be effective, the goal of ART should include the transfer of euploid embryos to maximize the potential for a healthy live birth. The objective of this study was to evaluate the clinical efficacy of blastocyst CCS with trophectoderm (TE) biopsy and vitrification in ART candidates of advanced maternal age (AMA). DESIGN: IRB approved randomized control trial. MATERIALS AND METHODS: Infertile patients of maternal age >35 years were computer randomized at oocyte retrieval into either: Group A 1⁄4 Fresh blastocyst transfer with embryos selected by morphology alone (n1⁄430) or Group B 1⁄4 Frozen blastocyst transfer with only euploid embryos tested by CCS (n1⁄430). CCSwas performed on TE biopsies using SNPmicroarray and all biopsied blastocysts were vitrified. RESULTS: There were no significant differences between the groups for maternal age (A1⁄439.7 2.2, B1⁄439.2 1.7 years), D3 FSH (A1⁄47.3 2.1, B1⁄46.8 2.1mIU/ml), AMH (A1⁄42.5 1.8, B1⁄42.7 2.1ng/ml) or AFC (A1⁄416.3 7.2, B1⁄420.9 10.1). 100% survival was observed following vitrification in Group B. Viable implantation rates (positive cardiac activity) (A1⁄440.9%, B1⁄460.8%; P<0.05) and the number of blastocysts transferred (A1⁄42.2 0.8, B1⁄41.7 0.5; P<0.01) were significantly different between the groups. In addition, significantly higher first trimester pregnancy losses were observed in Group A (A1⁄420%, B1⁄40%; P<0.05). CONCLUSION: Infertile AMA patients have higher ongoing implantation rates and fewer first trimester pregnancy losses, following a frozen blastocyst transfer with euploid embryos tested by CCS, when compared to routine fresh blastocyst transfer based on embryo morphology alone. Blastocyst CCS significantly increases the likelihood of ART success for AMA patients, which could allow for the realization of routine single blastocyst transfer, typically only recommended for younger good prognosis patients. Supported by: Ferring Pharmaceuticals.

Alteration of gene expression in human cumulus cells as a potential indicator of oocyte aneuploidy

Human female meiosis is particularly error prone, leading to the generation of aneuploidy, a problem which increases dramatically with advancing age. Despite being of great clinical importance, the genesis of oocyte aneuploidy remains poorly understood. Cumulus cells (CCs) surround oocytes in antral follicles and play a crucial role in regulating their maturation. During this investigation, we aimed to obtain an insight into the follicular environment of oocytes that become aneuploid and identify non-invasive markers of oocyte chromosome status and competence. Microarray comparative genomic hybridization was used to assess oocyte ploidy. Expression of 96 genes was examined via a large real-time PCR experiment in 51 CC samples, whereas an additional 26 CC clumps were used for a more focused real-time PCR experiment assessing just three genes. Gene selection was based on the results of a previous microarray analysis comparing the transcriptome of CCs from normal and aneuploid oocytes. Fifty eight of the investigated genes were confirmed to be expressed in CCs. Patterns of expression were generally similar among all CC samples, regardless of the chromosomal status of the corresponding oocyte. However, two genes, SPSB2 and TP53I3, were both significantly down-regulated in CCs from chromosomally abnormal oocytes (P<0.05). The expression of SPSB2 was also found to display a strong trend towards up-regulation in the CCs of oocytes that produced a healthy live birth (P=0.054). In the current study, SPSB2 and TP53I3 exhibited highly significant differences in their expression between CCs of normal and chromosomally abnormal oocytes. SPSB2 is involved in intracellular signalling and homeostasis, whereas TP53I3 regulates carbohydrate metabolism and apoptosis. We speculate that both these genes might have the potential to serve as non-invasive markers of oocyte aneuploidy.

Live birth after ovarian tissue autotransplantation following overnight transportation before cryopreservation

To describe the first live birth after transplantation of ovarian tissue following overnight transportation of the tissue before freezing. Technical note. University department of obstetrics and gynecology. A 25-year-old cancer survivor with previous Hodgkin disease and relapse. The ovarian tissue was kept cool for >20 hours in a special transport medium and a special cooling device before it was cryopreserved. After premature ovarian failure due to preconditioning chemotherapy for bone marrow transplantation, the cryopreserved ovarian tissue was transplanted orthotopically. Resumption of ovarian function after transplantation, recovery of fertility, and pregnancy. Ovarian function returned in the patient 3 months after transplantation, as shown by follicle development and estrogen production. During the fifth menstrual cycle, mild stimulation with FSH was initiated in accordance with a low-dose protocol. When ultrasonography revealed a follicle 18-20 mm in size in the ovarian graft, hCG was added and the patient had sexual intercourse at the optimal time point. On day 14 of the luteal phase, hCG concentration and vaginal echography confirmed a viable intrauterine pregnancy, which resulted in a healthy live birth. Overnight transportation of ovarian tissue appears to be possible in combination with appropriate transportation logistics. However, further investigations are needed before this procedure can be offered as a chance for women to preserve fertility independently of direct access to a tissue-processing bank.

Vitrification of human embryos previously cryostored by either slow freezing or vitrification results in high pregnancy rates

Occasionally, clinical scenarios arise where embryos, previously cryostored and warmed, need to be recryopreserved. The outcome of 30 such transfer cycles from 25 women where embryos were recryopreserved is detailed. In 16 cases, embryos were initially cryopreserved by slow freezing and in 14 cases by vitrification. The cryopreservation stages were the pronuclear stage (n=16), day-3 cleavage stage (n=12), blastocyst (n=1) and oocytes (n=1). All recryopreservation was by Cryotop-based vitrification. From this mixed source, 30/31 twice-cryopreserved embryos survived warming and were transferred, resulting in 13 pregnancies, 11 deliveries with normal gestational age and birthweight, one pre-term birth at 33 weeks and two miscarriages. There were no malformations reported for the live births. Recryopreservation using vitrification by CryoTop has been used in a variety of clinical scenarios to preserve surplus cryopreserved embryos. The current study, although limited in numbers, resulted in high survival rates, clinical pregnancy rates similar to once-cryopreserved embryos and healthy live births independently of the initial stage and cryopreservation method. The technique may increasingly be applicable to elective single-embryo transfer and blastocyst transfer to maximize the pregnancy rate while minimizing the number of cryopreserved embryo transfers.Freezing of embryos has significantly influenced the likelihood of pregnancy after an IVF stimulation cycle. Since it is often unclear which embryos have the potential to continue developing, more embryos may be thawed than transferred. If, in such cases, the surplus embryos remain viable, the ability to refreeze them is an important laboratory tool. In this study, we report on 30 cycles from 25 women where, for a variety of clinical scenarios, surplus embryos previously cryopreserved by either slow freezing or more recently by vitrification were recryopreserved using the Cryotop vitrification protocol. The report details 13 pregnancies, including 11 live births. The study group included one case where oocytes rather than embryos were initially vitrified. When these recryopreserved embryos were subsequently warmed, they exhibited a high survival rate and, after embryo transfer, a pregnancy rate similar to once-cryopreserved embryos. While recryopreservation has to date demonstrated that the embryos retain viability, leading to healthy children, and is, as such, an additional tool for embryologists, its application should proceed with caution.

Oocyte vitrification technology has made egg-sharing donation easier in China

When infertile women undergoing IVF or intracytoplasmic sperm injection (ICSI) have more than 20 mature oocytes retrieved, at least 15 oocytes are inseminated by their husband’s spermatozoa. The extra oocytes are cryopreserved by vitrification. If the patients became pregnant and have healthy live births, the patients are encouraged to donate their remaining cryopreserved oocytes. Forty-seven egg-sharing donors were recruited after having normal deliveries and they donated their remaining oocytes, totalling 395 cryopreserved oocytes, to 75 recipients. The survival rate of vitrified–warmed oocytes was 83.0%. Following insemination by ICSI, the fertilization and cleavage rates were 83.8% and 89.8%, respectively. Out of 75 recipients, 71 recipients completed the treatment cycles and 30 of them became pregnant with clinical pregnancy and implantation rates of 42.3% and 25.5%, respectively. The birthweight of the new-born infants (22 from singleton and two from one set of twins) were 3344.5 ± 669.1 g and 2425.0 ± 742.5 g, respectively. No birth defects were observed for the live births. These results indicate that oocyte vitrification is an effective methodology for an egg-sharing donation programme, with acceptable pregnancy and implantation rates. This study demonstrated that the strategy of egg-sharing donation with oocyte vitrification is a practical and efficient programme in China, especially with the strict regulations controlling egg-sharing donation. When infertile women undergoing IVF or intracytoplasmic sperm injection (ICSI) have more than 20 mature oocytes retrieved, at least 15 oocytes are inseminated by their husband’s spermatozoa. The extra oocytes are cryopreserved by vitrification. If the patients become pregnant and have healthy live births, the patients are encouraged to donate their remaining cryopreserved oocytes. Forty-seven egg-sharing donors were recruited after having healthy deliveries and they donated their remaining oocytes, in total 395 cryopreserved oocytes to 75 recipients. The survival rate of vitrified–warmed oocytes was 83.0%. Following insemination by ICSI, the fertilization and cleavage rates were 83.8% and 89.8% respectively. Out of 75 recipients, 71 recipients completed their treatment cycle and 30 of them became pregnant (42.3% clinical pregnancy rate and 25.5% implantation rate). The birthweight of new-born infants for 22 singletons and two babies from one set of twins were in the normal range. No birth defects were observed for those live births. These results indicate that oocyte vitrification is an effective methodology for an egg-sharing donation programme, and gives acceptable pregnancy and implantation rates.