Repetitive Implantation Failure Research Articles

The Current Role of Induced Endometrial Trauma (Endometrial Scratch) in Women Undergoing Infertility Treatment.

Induced endometrial trauma, otherwise known as endometrial scratch is a simple technique that has been rapidly adopted into clinical practice, mainly for women having IVF treatment, in an attempt to increase pregnancy rates. The introduction of endometrial scratch followed early reports of improved clinical pregnancy rates in women with repetitive implantation failure after having the procedure and follows on from evidence from animal models in the early 20th century suggesting that mechanical trauma to the endometrium can induce decidual changes. Due to the ease and low cost of the procedure, it has been rapidly adopted as an add-on to fertility treatments, in many cases where evidence is still lacking. Despite the initial publication of a large number of studies that demonstrated encouraging improvements in pregnancy rates in women who underwent this procedure, these studies were mainly limited by the small sample sizes and heterogeneity of their study populations, leading to limited validity of the evidence provided by these studies. More recently, three large randomized controlled studies have been published that paint a different picture regarding the value of this procedure. This article explores the evolution of the evidence and the current state of endometrial scratch as an adjuvant therapy for women undergoing IVF treatment.

Preimplantation Genetic Testing for Aneuploidy: Current Perspectives.

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.

PREIMPLANTATION GENETIC TESTING: Chromosome abnormalities in human embryos

Aneuploidy is a frequent event in human embryos, and its incidence is higher in oocytes and embryos from women of advanced maternal age. Aneuploidy may also be a contributing factor in infertile populations, such as couples with recurrent miscarriages, repetitive implantation failure, or male infertility. For these reasons, preimplantation genetic testing for aneuploidy (PGT-A) has been proposed to prevent miscarriages and increase live birth rates in infertile couples undergoing in vitro fertilisation. Next-generation sequencing is currently being applied for the detection of aneuploidies in human embryos, including whole chromosome aneuploidies, segmental aneuploidies, uniform, and mosaic aneuploidies. More recently, this technology has been incorporated for the analysis of the cell-free DNA secreted by the embryo to the culture media. Chromosome abnormalities mostly originate in female meiosis. Recombination between homologous chromosomes is a critical event that occurs in the foetal ovary. The importance of altered recombination pertains to paternally as well as maternally derived trisomies, but as most aneuploidy arises during oogenesis, the female is at greater risk. For males, sperm concentration is associated with a higher risk of aneuploid sperm and thus aneuploid embryos. Mitosis errors can occur at all stages of early embryo development that result in chromosomally distinct cell populations. The clinical impact of mosaicism depends on the mosaicism type, location, and number of aneuploid cells. Transfer of mosaic embryos has been proposed when no euploid embryos are available in the PGT-A cycle.

Pre‐implantation genetic testing for aneuploidy: the past, present and future

Key content Embryonic aneuploidy is often found in women undergoing assisted reproduction and is a common cause of poor outcomes, particularly in women of advanced maternal age and those with recurrent miscarriage and repetitive implantation failure. The aim of pre‐implantation genetic testing for aneuploidy (PGT‐A) is to select the most competent embryo for transfer, thus improving reproductive outcome. Initial PGT‐A techniques failed to show an improvement in delivery rates and, in some trials, demonstrated inferior outcomes; however, advances in technology, combined with robust culture media and vitrification techniques, have significantly improved outcomes. Recent debate has argued for the increased clinical application of PGT‐A, particularly in women of advanced maternal age; we provide an overview of PGT‐A, including its past and associated limitations, the evolution in technology and latest outcomes, and address potential future implications. Learning objectives To understand the rationale behind the use of PGT‐A. To review the evidence against and in favour of PGT‐A. To understand the new techniques being used and how these have led to emerging evidence in favour of PGT‐A. Ethical issues Risk of damage to embryos during biopsying process. PGT‐A may result in either no embryos available for transfer or only embryos displaying mosaicism being available to transfer, with little long‐term data on the outcomes of transferring mosaic embryos. PGT‐A increases the cost of an already expensive treatment.

Sperm chromosomal abnormalities and their contribution to human embryo aneuploidy.

In this work we reviewed 18years of experience using fluorescence in situ hybridization (FISH) for sperm aneuploidy testing. We evaluated parameters associated with increased numerical sperm chromosome abnormalities and determined the male contribution to embryo aneploidies in terms of reproductive outcome by increased sperm aneuploidy. This retrospective study analyzed data from 2008 sperm samples of infertile males undergoing FISH analysis because of clinical history of repetitive implantation failure, recurrent miscarriage, impaired sperm parameters, or mixed causes. Sperm concentration was the only sperm parameter associated with FISH results-we observed a gradual increase of abnormal sperm FISH results in males with decreasing sperm concentration. However, a great proportion of normozoospermic males also showed increased sperm aneuploidies, suggesting that sperm parameters alone do not enable identification of a substantial proportion of infertile males at risk of sperm aneuploidies. Regarding reproductive outcomes, couples with normal sperm FISH results for the male had similar outcomes regardless of conventional in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), or preimplantation genetic testing for aneuploidies (PGT-A). However, couples with abnormal sperm FISH results for the male showed better clinical outcomes after PGT-A, suggesting a potential contribution of sperm to embryo aneuploidy. Moreover, PGT-A cycles showed better clinical outcomes when 24 chromosomes were analyzed by array comparative genome hybridization (aCGH) or next-generation sequencing (NGS) instead of only nine chromosomes analyzed by FISH. In conclusion, sperm FISH analysis offers clinical prognostic value to evaluate reproductive possibilities in infertile couples. Therefore, couples with abnormal sperm FISH results should be offered genetic counseling and presented with clinical options such as PGT-A.

Is euploidy what embryos need for implantation and live birth? We also need quality

Introduction To have an euploid embryo is one of the main biomarkers conditioning the outcome of an IVF cycle since the sole morphological appearance it is not always related to the euploidy status of the embryo. Blastocysts with a good morphology can be aneuploid and vice versa. In this study, the implantation potential and the live birth rate (LBR) of euploid warmed blastocyst after PGT was analyzed according to the quality of the biopsied blastocysts. Additionally, the age factor was also considered. We evaluated the association between the patient´s age and the euploidy status of the embryos. Materials and Methods We included retrospectively 976 warmed blastocysts from a total of 828cycles, since January 2016 to December 2017, from our PGT program (1504-VLC-027-MD). Indications for PGT-A were advanced maternal age (66.8%), repetitive implantation failure (12.7%), recurrent miscarriage (7.0%), severe male factor (12.7%), previous pregnancy with aneuploidy (2.5%) and mixed causes (2.3%).Thetrophectoderm biopsy was completed either on Day 5 or on Day 6 of embryo development according to the degree of expansion and quality reached by the blastocyst. The trophectoderm analysis was done by NGS, and one hour after biopsy blastocysts were vitrified. The day of the transfer, only euploid embryos were warmed, waited for a minimal of two hour for their re- expansion and then graded according the morphological ASEBIR classification (type A, B, C), based on both the degree of expansion and the quality of the trophectoderm. Implantation and LBR were analyzed according to the type of embryo with a Chi-squared test. Statistical significance was achieved for values of p 38 years. Implantation potential of transferred blastocysts was compared between both groups. Results Weobtained statistical significant differences in implantation potential (p Conclusions For euploid embryos, the grading of the blastocysts will determine the implantation potential and the chances of a live birth. These differences are of crucial importance for biopsied blastocysts on Day 6. Regarding the age of the patient, once we have euploid embryos, the patients ‘age is not a decisive parameter in terms of chances of embryo´s implanting.

Clinical results and perinatal outcome in PGT-A cycles with trophectoderm biopsy: a 4-year follow up study

Introduction Trophectoderm biopsy is one of the strategies to obtain a sample to analyze the aneuploidies of the embryo. Blastocyst vitrification after biopsy give us more time for analysis. Moreover, it is a strategy that allows transfer back the normal embryo in a non-stimulated cycle. The endpoint of an assisted reproduction cycle is a healthy live newborn at home. Therefore, perinatal outcome follow-up is necessary. The objective of this study was to assess the results and perinatal outcome obtained in couples from our PGT-A program with trophectoderm biopsy. Material & Methods Retrospective collection data value (1503-VLC-017-AM) from our PGT-A program (2014-2017) and pregnancies and perinatal follow-up resulted from these cycles until September 2017. This study included 1,818 PGT-A cycles (39 in 2014, 98 in 2015, 693 in 2016 and 1,051 in 2017). Indications for PGT-A were advanced maternal age (64.8%), repetitive implantation failure (12.5%), recurrent miscarriage (7.2%), severe male factor (10.7%), previous pregnancy with aneuploidy (2.5%) and mixed causes (2.3%). Trophectoderm biopsy was carried out in fresh blastocysts. Biopsies were performed on day-5 or day-6 and one hour after biopsy blastocysts were vitrified until results. Analysis by NGS were performed. Only euploid blastocysts were transferred in a next cryotransfer. Data regarding to deliveries and babies were obtained from the medical records, as well as the reports sent by the referral center and from the survey sent to the patients. Results For cycles with almost one euploid blastocyst, the corresponding rates were 87.2% (n=39) in 2014, 71.4% (n=98) in 2015, 77.9% (n=693) in 2016 and 65.2% (n=1051) in 2017. Number of transferences were: 70 in 2014, 109 in 2015, 613 in 2016 and 695 in 2017. Known implantation rate and ongoing pregnancy were 64.9% and 48.9%, respectively. Number of double embryo transfer (DET) were high in 2014 (60.8%) and in 2015 (47.7%), and the multiple pregnancies remained high too (51.0%). Because the ongoing pregnancy rate in this years the transfer policy changed from DET to single embryo transfer (SET). In 2016 and 2017, SET represented 71.0% and 78.9% from the embryo transfers performed, respectively, decreasing the multiple pregnancy rates to 16.1%. Caesarean section was performed in 63.8% of the deliveries reported. Out of 336 children reported born, sex distribution was: 53.5% males and 46.5 females. Very premature delivery (before week 32) occurred in 2.5% of the pregnancies. A total of 336 babies were born. The average weigth was 3240±1200g and the average length was 50.1±4.9cm. Several congenital abnormalities were found that were not related to the PGT indication (less than 1%). Conclusion Our results suggest than trophectoderm biopsy is a safe option that does not compromise the clinical results and perinatal outcome in a PGT-A program. The annual data collection provide an important resource for data mining and for following trends in PGT practice, the same way ESHRE Consortium and ASRM recommend.

Clinical application of embryo aneuploidy testing by next-generation sequencing.

We review here the evolution in the field of embryo aneuploidy testing over the last 20 years, from the analysis of a subset of chromosomes by fluorescence in situ hybridisation to the transition toward a more comprehensive analysis of all 24 chromosomes. This current comprehensive aneuploidy testing most commonly employs next-generation sequencing (NGS). We present our experience in over 130 000 embryo biopsies using this technology. The incidence of aneuploidy was lower in trophectoderm biopsies compared to cleavage-stage biopsies. We also confirmed by NGS that embryo aneuploidy rates increased with increasing maternal age, mostly attributable to an increase in complex aneuploid embryos. In contrast, the number of MII oocytes retrieved or the use of oocyte vitrification did not affect aneuploidy rates. Similarly, neither maternal age, oocyte number, nor oocyte vitrification affected the incidence of mosaicism. Analysis of clinical outcomes, indications, and potential benefits of embryo aneuploidy testing revealed advanced maternal age as the most favored group, with some evidence of improved delivery rate per transfer as well as decreased miscarriage rates and time to pregnancy. Other indications are: recurrent miscarriage, repetitive implantation failure, severe male factor, previous trisomic pregnancy, and good prognosis patients mainly undergoing single embryo transfer, with the latter indication used to reduce the occurrence of multiple pregnancies without compromising cycle outcome. In conclusion, NGS has become the most appropriate technology for aneuploidy testing in trophectoderm biopsies, with accurate results, high throughput, and cost efficiency. This technology can be also applied to the analysis of the embryonic cell free DNA released to the culture media at blastocyst stage. This is a promising approach towards a non-invasive preimplantation genetic testing of aneuploidy.

Aneuploidy screening by array comparative genomic hybridization improves success rates of in vitro fertilization: A multicenter Indian study.

OBJECTIVE:To evaluate the usefulness of preimplantation genetic screening (PGS) using array comparative genomic hybridization (aCGH) in the Indian population.MATERIALS AND METHODS:This is a retrospective, multicenter study including 235 PGS cycles following intracytoplasmic sperm injection performed at six different infertility centers from September 2013 to June 2015. Patients were divided as per maternal age in several groups (<35, 35–36, 37–38, 39–40, and >40 years) and as per indication for undergoing PGS. Indications for performing PGS were recurrent miscarriage, repetitive implantation failure, severe male factor, previous trisomic pregnancy, and advanced maternal age (≥35). Day 3 embryo biopsy was performed and analyzed by aCGH followed by day 5 embryo transfer in the same cycle or the following cycle. Outcomes such as pregnancy rates (PRs)/transfer, implantation rates, miscarriage rates, percentage of abnormal embryos, and number of embryos with more than one aneuploidy and chaotic patterns were recorded for all the treated subjects based on different age and indication groups.RESULTS:aCGH helped in identifying aneuploid embryos, thus leading to consistent implantation (range: 33.3%–42.9%) and PRs per transfer (range: 31.8%–54.9%) that were obtained for all the indications in all the age groups, after performing PGS.CONCLUSION:Aneuploidy is one of the major factors which affect embryo implantation. aCGH can be successfully employed for screening of aneuploid embryos. When euploid embryos are transferred, an increase in PRs can be achieved irrespective of the age or the indication.

Can intracytoplasmic sperm injection prevent total fertilization failure and enhance embryo quality in patients with non-male factor infertility?

ObjectiveTo determine whether intracytoplasmic sperm injection (ICSI) could prevent total fertilization failure (TFF) and enhance the embryo quality in patients with non-male factor infertility. Study designA total of 296 in vitro fertilization (IVF) cycles performed in patients with non-male factor infertility between April 2009 and March 2013 were included in this retrospective study. During the period, ICSI and conventional IVF were performed in 142 and 154 cycles, respectively. The usual indications for ICSI were in the cycles of patients with (1) known low fertilization rate, (2) repetitive implantation failure, (3) advanced maternal age, (4) presence of endometrioma, (5) low oocyte yield (number of oocytes ≤3), or (6) poor quality oocytes. The rate of TFF, normal fertilization, abnormal pronuclei (PN) formation, embryo quality, and pregnancy outcomes between the patients treated with ICSI and conventional IVF cycles were compared. ResultsThe patients treated with ICSI (ICSI group, n=142) presented fewer number of oocytes than patients treated with conventional IVF cycles (n=154). The TFF rate was not different (4.2% vs. 0.6%, P=0.059), but the ICSI group presented a significantly higher rate of normal fertilization (83.4% vs. 79.1%, P=0.04) and lower rate of abnormal PN formation (3.9% vs. 13.3%, P<0.01). The cleavage stage embryo quality was better in the ICSI group (grade A: 31.1% vs. 21.3%, P=0.001; grade A+B: 65.1% vs. 47.6%, P<0.001). ConclusionThe result of this study does not support the use of ICSI to prevent TFF in patients with non-male factor infertility. However, ICSI improved the fertilization rate and the embryo quality.

New tools for embryo selection: comprehensive chromosome screening by array comparative genomic hybridization.

The objective of this study was to evaluate the usefulness of comprehensive chromosome screening (CCS) using array comparative genomic hybridization (aCGH). The study included 1420 CCS cycles for recurrent miscarriage (n = 203); repetitive implantation failure (n = 188); severe male factor (n = 116); previous trisomic pregnancy (n = 33); and advanced maternal age (n = 880). CCS was performed in cycles with fresh oocytes and embryos (n = 774); mixed cycles with fresh and vitrified oocytes (n = 320); mixed cycles with fresh and vitrified day-2 embryos (n = 235); and mixed cycles with fresh and vitrified day-3 embryos (n = 91). Day-3 embryo biopsy was performed and analyzed by aCGH followed by day-5 embryo transfer. Consistent implantation (range: 40.5–54.2%) and pregnancy rates per transfer (range: 46.0–62.9%) were obtained for all the indications and independently of the origin of the oocytes or embryos. However, a lower delivery rate per cycle was achieved in women aged over 40 years (18.1%) due to the higher percentage of aneuploid embryos (85.3%) and lower number of cycles with at least one euploid embryo available per transfer (40.3%). We concluded that aneuploidy is one of the major factors which affect embryo implantation.

Preimplantation genetic screening using fluorescence in situ hybridization in patients with repetitive implantation failure and advanced maternal age: two randomized trials

To evaluate the usefulness of preimplantation genetic screening (PGS) using fluorescence in situ hybridization (FISH) for two different indications: repetitive implantation failure (RIF) and advanced maternal age (AMA). Two prospective, randomized controlled trials with patients allocated in two arms: blastocyst transfer on day 5 (group A) or PGS with transfer on day 5 (group B). University-affiliated private clinics. The RIF study included women <40 years with three or more failed IVF cycles without other known causal factors (91 patients). The AMA study included intracytoplasmic sperm injection patients aged between 41 and 44 (183 patients). In the PGS group, single-cell day 3 biopsy was performed with aneuploidy screening for chromosomes 13, 15, 16, 17, 18, 21, 22, X, and Y. In both the blastocyst transfer group and the PGS group, ET was performed on day5. Live-birth rate per patient and per started cycle. A significant increase in live-birth rates per patient was found in the PGS group compared with the blastocyst group for the AMA study (30/93 patients [32.3%] vs. 14/90 patients [15.5%]; odds ratio, 2.585; confidence interval, [1.262-5.295]). In the RIF study no significant differences were observed (23/48 patients [47.9%] vs. 12/43 patients [27.9%]). PGS with FISH was shown to be beneficial for the AMA group.

Comparison of day-3 and day-5 array-CGH diagnosis for 24 chromosome aneuploidy screening in terms of accuracy

Comparison of day-3 and day-5 array-CGH diagnosis for 24 chromosome aneuploidy screening in terms of accuracy

High pregnancy rates with administration of granulocyte colony-stimulating factor in ART-patients with repetitive implantation failure and lacking killer-cell immunglobulin-like receptors

Sir, As early as the year 2000, we reported on the use of granulocyte colony-stimulating factor (G-CSF) for IVF patients who failed to become pregnant after repeated embryo transfers (Wurfel, 2000; Wurfel, 2003). At that time, we administered 300 µg of Molgramostin in a single dose on the day of the embryo transfer. The pregnancy rate of patients treated with transfer on day 2 (average two embryos) was almost 43% compared with almost 20% in the placebo group. The study by Scarpellini et al. (2009) now reports on continuous administration of G-CSF, evidently without any identifiable negative effects on the infants. Given this, we decided to run a pilot study to investigate whether continuous administration of G-CSF to patients with RIF (repetitive implantation failure) would be beneficial or not. However, the initial criterion—patients' failure to conceive after repeated IVF or ICSI treatments—appeared to us to be too vague. Taking the publication by Hiby et al. (2008) as a basis we initially performed killer-cell immunglobulin-like receptor (KIR) typing on patients with more than five unsuccessful IVF treatments or embryo transfers. Only patients who lacked the three activating receptors were accepted for the study; also included were patients suffering from long-term unexplained sterility (average 6.8 years) and lacking the three activating KIR genes. Groups overlapped to some extent as cases of unexplained sterility had often undergone (unsuccessful) IVF treatment. The incidence of three lacking receptors (2 DS 1, 2 DS 3 and 3 DS 5) was very high in the group examined, at 78%. In addition, it was noted that patients lacked not only the three named activating receptors but also further receptors, so that in general the group was shown to lack five to seven receptors including the above-mentioned three activating receptors. We performed IVF and ICSI treatment on the defined group with the target of a day 5 transfer. Patients received 13 million units of GranocyteTM (Lanogrostim) every 3 days in addition to the usual luteal support. In cases where insufficient numbers of fertilized oocytes were available, the transfer was performed on day 2 (particularly in cases of ovarian insufficiency). Results for d + 5: Exclusively transfer of two blastocysts/morulae or compacted morulae Number of patients 40 Number of treatment cycles 42 Clinical pregnancies (excluding biochemical) 31 of which abortions (clinical) 12 Pregnancy rate per treatment cycle/embryo transfer 73.8% Abortion rate (clinical pregnancies) 38.7% View it in a separate window Results for transfer d + 2 (also including patients with significant ovarian insufficiency) Number of patients 19 Number of treatment cycles 19 Clinical pregnancies 8 of which abortion (clinical) 3 Pregnancy rate per embryo transfer 42% Abortion rate 37.5% View it in a separate window The group we selected and defined certainly had a very poor prognosis. The pregnancy rates achieved were extraordinarily high both in the day 5 transfers and the day 2 transfers. However, the rate of clinical abortions is also high (biochemical pregnancies were ignored). In the meantime, we have also conducted a further pilot study in which G-CSF was administered to patients with a history of multiple unsuccessful IVF treatments or unexplained sterility, who had no KIR defects. The results were very poor, with pregnancy rates currently below 10% per embryo transfer. We conclude from this that the use of G-CSF is an extremely promising additional method of treatment in cases where defects in materno-embryonic implantation communication can be shown. This applies in particular to KIR defects and, in this, particularly to the lack of the three activating receptors as described by Hiby et al. Where such defects were not present, results of G-CSF treatment were disappointing. On the basis of the results of these pilot studies, we are currently planning a prospective randomized double-blind study under the initial criteria given above.

Reply: High pregnancy rates with administration of granulocyte colony-stimulating factor in ART patients with repetitive implantation failure and lacking killer-cell immunglobulin-like receptors

Reply: High pregnancy rates with administration of granulocyte colony-stimulating factor in ART patients with repetitive implantation failure and lacking killer-cell immunglobulin-like receptors

Polar body genetic diagnosis (PbGD) is a useful tool to improve the results of assisted reproductive technology (ART) and helps couples in their reproductive decisions

OBJECTIVE: Preimplantational Genetic Diagnosis is a successful tool to identify specific genetic abnormalities in embryos. It has also been used to screen aneuploid embryos in women ≥38 and patients with repetitive implantation failure. In communities where it is not acceptable to discard embryos, we have previously demonstrated that Polar Body Genetic Diagnosis (PbGD) is a reasonable tool for the selection of abnormal oocytes. The aim of the present study is to determine whether PbGD improves ART results and whether it is a useful diagnostic tool to help couples take reproductive decisions.

Births after transfer of zona-free blastocysts in oocyte donation cycles.

Oocyte donation is a well-established method of assisted reproduction for women with irreversible infertility and with previous implantation failures after in vitro fertilization. Although the pregnancy rates are very high, sometimes implantation does not occur even after various attempts. We report the first two cases of transfer of zona-free blastocysts in oocyte donation cycles that developed to normal pregnancies and births. The patients had undergone three previous standard oocyte donation cycles with failure of implantation. Endometrium preparation was performed after suppression of the pituitary function, with E2 valerate and Progesterone at the day of oocyte retrieval. Normally fertilized embryos were cultured in Earle's culture medium until Day 3 and in S2 medium until Day 5. For each patient, the zonae of two fully expanded blastocysts were enzymatically removed with 0.5% pronase. Zona-free blastocysts were transferred for the patients 2 h later. On Day 12 after transfer, pregnancies were confirmed with elevated serum levels of beta hCG. A gestational sac with a foetal heart beat was seen by ultrasound 15 days later, in each patient. Normal healthy babies were born at 38 and 39 weeks of pregnancy. This is the first report of successful pregnancies and births after oocyte donation and transfer of zona-free blastocysts in human. It not only shows the feasibility of the treatment but also opens a new alternative for the patients with repetitive implantation failure after OD cycles.

Fertilization and early embryology: Granulosa cells improve human embryo development in vitro

A total of 17 couples with repetitive implantation failure after transfer of fresh or frozen-thawed embryos had half of their zygotes cultured in standard conditions and frozen at day 2 after insemination, and the other half cocultured with autologous granulosa cells and transferred at the morula or blastocyst stage at day 5 or 6 after oocyte retrieval. At the end of the culture period, supernatants of cocultures were recovered for steroid assays. Monolayers were stained for granulosa cell growth and morphological assessment. We observed that granulosa cells improve embryo development in vitro since 32 out of 60 (53%) reached the morula stage and 18 (30%) the blastocyst stage, leading to a total of 83% embryos available for transfer (compared with 3% without coculture). The ongoing pregnancy rate of these patients who were selected because they had at least three previous implantation failures, is only 5.9%, however, which is similar to the control group without coculture (6.3%). To conclude, granulosa cells improve embryo development but not the pregnancy rate after transfer of cocultured embryos in patients with multiple previous implantation failures.