Regular IVF Research Articles

307 Pregnancy rate after icsi compared to a low-cost simplified ivf system in a selected population: A prospective cohort study

Aim of the study: By using split ICSI/simplified IVF culture system (SCS) cycles in a selected patient cohort, we examined if regular IVF methodology could be significantly simplified and still yields results comparable with ICSI. Methods: This analysis is part of a non-inferiority prospective study comparing ICSI and SCS for gamete fertilization and embryo culture in a selected group of patients. For women, selection criteria included being under 43 years of age and to have at least 6 eggs at retrieval.

Exploration of the Cytoplasmic Function of Abnormally Fertilized Embryos via Novel Pronuclear-Stage Cytoplasmic Transfer.

In regular IVF, a portion of oocytes exhibit abnormal numbers of pronuclei (PN) that is considered as abnormal fertilization, and they are routinely discarded. However, it is known that abnormal ploidy still does not completely abandon embryo development and implantation. To explore the potential of cytoplasm from those abnormally fertilized oocytes, we developed a novel technique for the transfer of large cytoplasm between pronuclear-stage mouse embryos, and assessed its impact. A large volume of cytoplast could be efficiently transferred in the PN stage using a novel two-step method of pronuclear-stage cytoplasmic transfer (PNCT). PNCT revealed the difference in the cytoplasmic function among abnormally fertilized embryos where the cytoplasm of 3PN was developmentally more competent than 1PN, and the supplementing of fresh 3PN cytoplasm restored the impaired developmental potential of postovulatory “aged” oocytes. PNCT-derived embryos harbored significantly higher mitochondrial DNA copies, ATP content, oxygen consumption rate, and total cells. The difference in cytoplasmic function between 3PN and 1PN mouse oocytes probably attributed to the proper activation via sperm and may impact subsequent epigenetic events. These results imply that PNCT may serve as a potential alternative treatment to whole egg donation for patients with age-related recurrent IVF failure.

Gonadotrophin stimulation and risk of relapse in breast cancer.

STUDY QUESTIONIs gonadotrophin stimulation as part of IVF associated with an increased risk of relapse in breast cancer?SUMMARY ANSWERControlled ovarian stimulation (COS) in connection with IVF in women with previous breast cancer was not associated with an increased risk of breast cancer relapse.WHAT IS KNOWN ALREADYBreast cancer is the most common malignancy among women worldwide and the leading cause of cancer death among females. The use of COS with gonadotrophins with subsequent cryopreservation of oocytes or embryos in order to enhance the chances of pregnancy after cancer treatment is the current most established fertility preservation method for women with breast cancer. To date, there are only a few small retrospective hospital-based controlled studies evaluating the risk of breast cancer relapse in patients undergoing fertility preservation with or without COS, showing no evident risk of relapse in breast cancer after the use of gonadotoxic agents.STUDY DESIGN, SIZE, DURATIONThis was a retrospective, population-based cohort study comprising 5857 women with previous breast cancer of whom 337 were exposed to COS. Exposure (COS) and outcomes (relapse and death) were identified for all patients from 2005 to 2014 by assessing the National Quality Register for Assisted Reproduction, the Swedish Medical Birth Register, the National Patient Register, the Swedish Prescribed Drug Register, the Swedish Cause of Death Register, the National Breast Cancer Register and the Swedish Cancer Register. Matching according to set criteria was possible for 334 women, who constituted the control group. A total of 274 women had undergone IVF after completing breast cancer treatment and 63 women had undergone COS for fertility preservation at the time of breast cancer diagnosis.PARTICIPANTS/MATERIALS, SETTING, METHODSWomen aged 20–44 years previously diagnosed with breast cancer and exposed to COS were matched for age at breast cancer diagnosis ±5 years, tumour size and lymph node involvement with a non-exposed control group, including women with known T- and N-stages. In a subsequent analysis, the matched cohort was assessed by also including women with unknown T- and N-stages. A secondary analysis comprised the entire non-matched cohort, including all women with known T- and N-stages. Also here, a subsequent analysis included women with missing data for T- and N-stages. The risk of relapse in breast cancer was estimated as crude hazard ratios (HRs) and 95% CI using Cox proportional hazards models in the primary and secondary analyses where T- and N-stages were known: otherwise the risks of relapse were only given descriptively.MAIN RESULTS AND THE ROLE OF CHANCEIn the primary matched analysis, relapse occurred in 20 of 126 women exposed to COS (15.9%) compared with 39 of 126 (31.0%) in the control cohort (HR = 0.70; 95% CI 0.39–1.45; P = 0.22). In the subsequent analysis, also including women with unknown T- and N-stages, relapse occurred in 27 of 337 (8.0%) women having undergone COS compared with 71/334 (21.3%) among the non-exposed. In the secondary adjusted analysis, relapse occurred in 20 of 126 (15.9%) exposed women and in 918 of 3729 (24.6%) non-exposed women (HR = 0.81; 95% CI 0.49–1.33; P = 0.70). In the subsequent analysis, including unknown T- and N-stages, relapse occurred in 27 of 337 (8.0%) women in the exposed group and 1176 of 5520 (21.3%) in the non-exposed cohort.LIMITATIONS, REASONS FOR CAUTIONA substantial degree of missing data on important prognostic variables was a limitation, particularly when analysing the total cohort. Furthermore, data on confounding factors, such as BMI, were not completely covered. Another limitation was that a pre-specified variable for relapse was not in use for the majority of the National Breast Cancer Register. Furthermore, the follow-up time from available register data (2005–2014) is rather short. Finally, we cannot be sure whether the prognostic information from receptor status, showing a lower incidence in the exposed group, is representative. Information on T- and N-stages was missing in more than half of the patients.WIDER IMPLICATIONS OF THE FINDINGSIn this large, retrospective, matched cohort study, we found no increased risk of relapse in breast cancer among women who had been exposed to gonadotrophins as part of IVF. This is reassuring but might be confounded by the selection of a group of women with a more favourable prognosis than those not undergoing IVF. The present study strengthens previous findings by being large, national and register based. Its results are applicable to women undergoing fertility preservation as well as to those undergoing regular IVF treatment.STUDY FUNDING/COMPETING INTEREST(S)Supported in part by grants from the Swedish state under the agreement between the Swedish government and the county councils the ALF-agreement (ALFGBG-720291), The Assar Gabrielsson Fund (FB 15-20), The Breast Cancer Fund and the Swedish Association of Local authorities and Regions, SKR. There are no conflicts of interest to declare.TRIAL REGISTRATIONN/A

PGT-A in Poor, Normo- and Hyper-Responders

The number of embryos available for transfer is significantly lower in PGT-A than in regular IVF simply because of genetic selection. Our aim is to evaluate ovarian reserve and ovarian response in relation to the probability of finding at least one euploid embryo (POE), which is the main determinant for PGT-A success. Over a six-year period, 2011-2017, PGT-A cycles were retrospectively analyzed and 5952 blastocysts were biopsied and diagnosed either by aCGH or NGS. The POE per cycle and the euploidy rate (ER) per diagnosed embryos were evaluated according to maternal age and ovarian reserve as measured by Anti Mullerian Hormone (AMH), day3 Follicle Stimulating Hormone (FSH) levels, and ovarian response as measured by the number of cumulus-oocyte complexes (COCs), metaphase II (MII) oocytes retrieved, top and good quality blastocyst (TQ+GQ) and biopsied blastocysts. According to a logistic regression analysis conducted to correlate maternal age and the ER per cycle, ER decreased by 1.9-fold with each additional year, whereas for POE, the same decline per cycle was around 0.8-fold in a generalized linear mixed model (GLMM) (p In conclusion female age is the most significant predictor of euploidy rate.Euploidy rate does not directly correlate with the number of oocytes retrieved. POE decreases significantly with advancing maternal age. AMH level, d3 FSH levels, the number of COCs, MII oocytes and the number of TQ+GQ and biopsied blastocyst are the main variables affecting POE after adjustment for age indicating that ovarian reserve and response have a significant impact. These findings emphasize the importance of evaluating and counselling the patients for factors both negative and positive, which may affect the success of PGT.

Impact of ovarian reserve on PGD

The number of embryos available for transfer is significantly lower in preimplantation genetic diagnosis (PGD) than in regular IVF because of genetic selection. Thus, the clinical evaluation of a patient applying for PGD is important in order to maximize the possibility of transferring a euploid embryo. The objective of the study was to determine the impact of ovarian reserve on PGD.

Growth factors IGF-I and IGF-II in human embryo co-culture with autologous granulosa cell clusters compared to regular embryo culture in IVF

To investigate and compare the presence of IGF-I and IGF-II in human embryo culture with and without autologous granulosa cell supplementation. Randomized prospective comparative study. 19 IVF couples agreed to participate in this study. For autologous comparison, half of the MII-oocytes from the same patient were assigned randomly either to co-culture (group I) or to regular culture (group II), treated for ICSI and cultured in microdrops of Irvine Continuous Single Culture Medium with 15% SSS. As control, autologous granulosa cell clusters were cultured alone (group C). On day 3, 50μl of supernatants from group I, II and control group C were collected and stored frozen. Supernatants were analyzed for IGF-I and IGF-II with growth factor-specific magnetic bead panels by Lab Supplies Inc., Athens, Greece. IGF-I was not detectable in group II, although it was detected in considerable levels in group I and in the control group C. The difference in concentration of IGF-I between groups I and C was not statistically significant (table 1). IGF-II was detected in all groups (see Table 1). However, IGF-II concentration in group II was at significantly higher levels when compared to group I and group C (p < 0.05 and < 0.04 respectively, two-tailed unpaired t-tests). We identified IGF-I both in co-culture and control supernatants but not in regular IVF culture, confirming that IGF-I is secreted from granulosa cells. We now propose that IGF-I is another essential growth factor for embryo culture. Investigators have in the past shown an anti-apoptotic effect of IGF-I on cell growth and morphology when added separately in embryo culture media without co-culture. Separately, we noted that IGF-II is present in samples from all groups, but excessively higher in samples from regular IVF culture, thus suggesting that granulosa cells regulate the level of IGF-II in the co-culture. We have previously documented that granulosa-cell supplementation to human embryo culture improves embryo development, due to beneficial contribution of growth factors (Ref. 1). Supplementation of embryo culture with granulosa cell clusters is therefore recommended for IVF.Tabled 1Tabel 1. Mean Concentration (pg/mL) of IGF-I and IGF-II in the three groups.Growth FactorGroup IGroup IIGroup CIGF-I11026.5IGF-II13.36624.3 Open table in a new tab

Impact of sperm DNA chromatin in the clinic.

The paternal contribution to fertilization and embryogenesis is frequently overlooked as the spermatozoon is often considered to be a silent vessel whose only function is to safely deliver the paternal genome to the maternal oocyte. In this article, we hope to demonstrate that this perception is far from the truth. Typically, infertile men have been unable to conceive naturally (or through regular IVF), and therefore, a perturbation of the genetic integrity of sperm heads in infertile males has been under-considered. The advent of intracytoplasmic sperm injection (ICSI) however has led to very successful treatment of male factor infertility and subsequent widespread use in IVF clinics worldwide. Until recently, little concern has been raised about the genetic quality of sperm in ICSI patients or the impact genetic aberrations could have on fertility and embryogenesis. This review highlights the importance of chromatin packaging in the sperm nucleus as essential for the establishment and maintenance of a viable pregnancy.

Low Ovarian Stimulation Using Tamoxifen/FSH Compared to Conventional IVF: A Cohort Comparative Study in Conventional IVF Treatments

Background: Over the last decade, the laboratory procedures in artificial reproduction have improved, thus fewer eggs are necessary. Therefore mild stimulations have been introduced to reduce the risk for the patient and patient drop-outs in treatment. The present study was designed to evaluate the efficacy of low stimulation with Tamoxifen and FSH in a routine IVF clinic. Material and methods: A total of 2,709 regular short antagonist IVF cycles were compared to 170 Tamoxifen low stimulation IVF cycles. All patients were recruited in the same time period and allocated to the different treatments on their own request. Results: No differences in age and number of previous cycles were found in between the groups. The clinical pregnancy rate was found to be significantly lower in the Tamoxifen treated group (20% vs. 26%), however we did not find any significant pregnancy rate following transfer (31% Tamoxifen group vs. 28% in conventional IVF). We found lower numbers of visits, lower costs for medication, less side effects and better acceptance for treatment in the Tamoxifen group. Conclusion: Although we found a lower pregnancy rate per started cycle, the pregnancy rate per transfer was equal and better accepted by the patients.

Safety, efficacy and efficiency of laser-assisted IVF in subfertile mutant mouse strains

In the present report we studied the safety, efficacy and efficiency of using an infrared laser to facilitate IVF by assessing fertilization, development and birth rates after laser-zona drilling (LZD) in 30 subfertile genetically modified (GM) mouse lines. We determined that LZD increased the fertilization rate four to ten times that of regular IVF, thus facilitating the derivation of 26 of 30 (86.7%) GM mouse lines. Cryopreserved two-cell stage embryos derived by LZD-assisted IVF were recovered and developed to blastocysts in vitro at the same rate as frozen-thawed embryos derived by regular IVF. Surprisingly after surgical transfer to pseudopregnant recipients the birth rate of embryos derived by LZD-assisted IVF was significantly lower than that of embryos derived by regular IVF. However this result could be completely mitigated by the addition of 0.25 M sucrose to the culture medium during LZD which caused the oocyte to shrink in volume relative to the perivitelline space. By increasing the distance from the laser target site on the zona pellucida, we hypothesize that the hyperosmotic effect of sucrose reduced the potential for laser-induced cytotoxic thermal damage to the underlying oocytes. With appropriate preparation and cautious application, our results indicate that LZD-assisted IVF is a safe, efficacious and efficient assisted reproductive technology for deriving mutant mouse lines with male factor infertility and subfertility caused by sperm-zona penetration defects.

Relationship quality in lesbian and heterosexual couples undergoing treatment with assisted reproduction

BACKGROUNDOne of the major factors impacting on a couple's relationship is the desire to have children. To many couples having a child is a confirmation of their love and relationship and a means to deepen and develop their intimate relationship. At the same time parental stress can impact on relationship quality. Relationship quality in lesbian couples is, currently, sparsely studied. The aim of the present study was to compare lesbian and heterosexual couples' perceptions of their relationship quality at the commencement of assisted reproduction, and to relate this to background data such as educational level, having previous children and, for lesbian couples, the use of a known versus anonymous donor.METHODSThe present study is part of the prospective longitudinal ‘Swedish study on gamete donation’, including all fertility clinics performing donation treatment in Sweden. Of a consecutive cohort of 214 lesbian couples about to receive donor insemination and 212 heterosexual couples starting regular IVF treatment, 166 lesbian couples (78% response) and 151 heterosexual couples (71% response) accepted participation in the study. At commencement of assisted reproduction participants individually completed questionnaires including the instrument ‘ENRICH’, which is a standardized measure concerning relationship quality.RESULTSIn general, the couples rated their relationship quality as good, the lesbian couple better than the heterosexuals. In addition, the lesbian women with previous children assessed their relationship quality lower than did the lesbian woman without previous children. For heterosexual couples previous children did not influence their relationship quality. Higher educational levels reduced the satisfaction with the sexual relationship (P = 0.04) for treated lesbian women, and enhanced the rating of conflict resolution for treated lesbian women (P = 0.03) and their partners (P = 0.02). Heterosexual women with high levels of education expressed more satisfaction with communication in their relationship (P = 0.02) than did heterosexual women with lower educational levels.CONCLUSIONSIn this Swedish study sample of lesbian and heterosexual couples' relationships, we found that they were generally well adjusted and stable in their relationships when starting treatment with donated sperm or IVF, respectively. However, where lesbian women had children from a previous relationship, it decreased relationship quality. For the heterosexual couples previous children did not affect relationship quality.

A case–control pilot study of low-intensity IVF in good-prognosis patients

Low-intensity IVF (LI-IVF) is rapidly gaining in popularity. Yet studies comparing LI-IVF to standard IVF are lacking. This is a case–control pilot study, reporting on 14 first LI-IVF and 14 standard IVF cycles in women with normal age-specific ovarian reserve under age 38, matched for age, laboratory environment, staff and time of cycle. LI-IVF cycles underwent mild ovarian stimulation, utilizing clomiphene citrate, augmented by low-dose gonadotrophin stimulation. Control patients underwent routine ovarian stimulation. LI-IVF and regular IVF patients were similar in age, body mass index, FSH and anti-Müllerian hormone. Standard IVF utilized more gonadotrophins (P<0.001), yielded more oocytes (P<0.001) and cryopreserved more embryos (P<0.001). With similar embryo numbers transferred, after ethnicity adjustments, standard IVF demonstrated better odds for pregnancy (OR 7.07; P=0.046) and higher cumulative pregnancy rates (63.3% versus 21.4%; OR 6.6; P=0.02). Adjustments for age, ethnicity and diagnosis maintained significance but oocyte adjustment did not. Cost assessments failed to reveal differences between LI-IVF and standard IVF. In this small study, LI-IVF reduced pregnancy chances without demonstrating cost advantages, raising questions about its utility. In the absence of established clinical and/or economic foundations, LI-IVF should be considered an experimental procedure.Low-intensity IVF (LI-IVF) is increasingly propagated as an alternative to standard IVF. LI-IVF has, however, never been properly assessed in comparison to standard IVF. Such a comparison is presented in the format of a small pilot study, matching LI-IVF cycles with regular IVF cycles and comparing outcomes as well as costs. The study suggests that LI-IVF, at least in this setting, is clinically inferior and economically at best similar to standard IVF. LI-IVF should, therefore, as of this point not be offered as routine IVF treatment but only as an experimental procedure.

285 THE EFFICIENCY OF INTRACYTOPLASMIC SPERM INJECTION v. LASER-ASSISTED IN VITRO FERTILIZATION WITH FROZEN SPERM FOR RECOVERY TRANSGENIC C57BL/6 MOUSE STRAINS

The progress of molecular genetics generated thousands of new transgenic strains of mice which also requires their economic and safe maintenance in the form of genetic banks. Sperm freezing would be one of the easiest options; however, cryosensitivity of sperm in mice strains is prone to variation. In this study, we examined the efficiency of laser-assisted zona drilling in vitro fertilization (ZD-IVF) v. intracytoplasmic sperm injection (ICSI) for attempting to recover two transgenic (UBI-GFP/BL6 and B6;129P2- Hvcn1) and one mutant (C57BL/6J-Tyrc-2J) lines. The sperm was frozen with 18% raffinose and 3% skim milk (Nakagata 2000 Mamm Genome 11, 572–576). Data of the replicates was analysed by chi-square method. The motility rates after thawing of cryopreserved sperm were 10% for UBI-GFP/BL6, 30% C57BL/6J-Tyrc-2J and 50% for B6;129P2-Hvcn1 strains. Regular IVF attempts in the UBI-GFP/BL6 and the mutant strain resulted in very few embryos and no pups (data not shown). Following ZD-IVF, the 2-cell stage rates were 7/60 (12%) for UBI-GFP/BL6, 18/60 (30%) for C57BL/6J-Tyrc-2J, and 34/60 (56%) for B6;129P2- Hvcn1. After ICSI, 66–74% of the oocytes survived the procedure and their development to 2-cells stage were 12/20 (60%) (UBI-GFP/BL6), 25/39 (64%) (C57BL/6J-Tyrc-2J) and 26/37 (70%) (B6;129P2-Hvcn1). The number of 2-cell stage embryos produced by ICSI was significantly (P &lt; 0.05) increased compared with those produced following ZD-IVF in case of UBI-GFP/BL6 and C57BL/6J-Tyrc-2J strains. The 2-cell stage embryos were transferred into recipients and the newborn rates from ZD-IVF v. ICSI embryos were 0% v. 17% (UBI-GFP/BL6), 28% v. 36% (C57BL/6J-Tyrc-2J) and 12% v. 12% (B6;129P2- Hvcn1), respectively; none of them were significantly different. In conclusion, when using cryopreserved sperm, the post-thaw motility is an important indicator for the selection of the rederivation method of cryopreserved transgenic mouse strains; while ZD-IVF, an easier method to perform, is suitable for the higher motility samples, ICSI could be strongly recommended for those showing low motility. This work was financed by EU FP6: CLONET (MRTN-CT-2006-035468), TEAMOHOLIC (MEXT-CT-2003-509582); EU FP7: RESOLVE (FP7-HEALTH-F4-2008-202047), RabPStem (PERG07-GA-2010-268422), and NKFP_07_1-ES2HEART-HU (OM-00202-2007).

Ovarian stimulation for fertility preservation reaches similar outcomes than normal IVF patients. an age-matched cases and controls study

OBJECTIVE: To evaluate the outcome of patients undergoing ovarian stimulation to preserve their fertility due to malignant diseases through oocyte vitrification in comparison with the outcome in regular IVF patients.DESIGN: Cases and controls, age-matched study.MATERIALS AND METHODS: Between 2008 and 2010, 74 patients were stimulated for oocyte vitrification. Ovarian stimulation results from patients with normal IVF women with an age difference <2 years and severe male factor (SMF) infertility matched by case, employed as controls. SMF was defined as having <100.000 sperm/ mL or azoospermia. Before starting any treatment, the oncologist agreed. Normal IVF patients were stimulated with GnRH analogues, agonist (long protocol) or antagonist (flexible protocol) and FSH with starting doses of 150-300 U/d. Patients with malignancies with receptors to steroids hormones were stimulated with letrozole 5mg daily added to FSH (150-300 U/d) (Gonal F, Merck-Serono, Spain) and antagonist of cetrorelix 3 mg (Cetrotide, Merck-Serono, Spain) when the leader follicle reaches 14 mm of mean diameter. Student's T was used to compare means between groups. CI denotes 95% confidence intervals.RESULTS: Cancer was in 48 patients from breast, 12 Hodgkin's disease, 4 ovarian, 4 colon, 2 Non-Hodgkin's disease, 1 bone and 3 with other malignancies.Age (31.7(CI30.4-33.0) vs. 32.4(CI31.3-33.5)), days of stimulation (10.1(CI9.6-10.6) vs. 10.4(CI9.9-10.7)), FSH doses (1472.2(CI(1342.4-1603.0) vs. 1630.5(CI1504.8-1756.2), oocytes retrieved (11.1(CI9.4-12.9) vs. 12.6(CI11.2-14.0), and mature oocytes (8.2(CI6.8-9.6) vs. 9.7(CI8.5-11.0) were statistically comparable.There were differences in E2 on the hCG day (pg/mL) being lower in the cancer group 803.4 (CI539.7-1067.1) vs. 2229.1(CI1923.6-2534.6).CONCLUSION: Similar outcome regarding the number of mature oocytes is achieved by patients with malignancies when compared with normal IVF patients. Ovarian stimulation with letrozole produces on them lower levels of estradiol at hCG day than other protocols. OBJECTIVE: To evaluate the outcome of patients undergoing ovarian stimulation to preserve their fertility due to malignant diseases through oocyte vitrification in comparison with the outcome in regular IVF patients. DESIGN: Cases and controls, age-matched study. MATERIALS AND METHODS: Between 2008 and 2010, 74 patients were stimulated for oocyte vitrification. Ovarian stimulation results from patients with normal IVF women with an age difference <2 years and severe male factor (SMF) infertility matched by case, employed as controls. SMF was defined as having <100.000 sperm/ mL or azoospermia. Before starting any treatment, the oncologist agreed. Normal IVF patients were stimulated with GnRH analogues, agonist (long protocol) or antagonist (flexible protocol) and FSH with starting doses of 150-300 U/d. Patients with malignancies with receptors to steroids hormones were stimulated with letrozole 5mg daily added to FSH (150-300 U/d) (Gonal F, Merck-Serono, Spain) and antagonist of cetrorelix 3 mg (Cetrotide, Merck-Serono, Spain) when the leader follicle reaches 14 mm of mean diameter. Student's T was used to compare means between groups. CI denotes 95% confidence intervals. RESULTS: Cancer was in 48 patients from breast, 12 Hodgkin's disease, 4 ovarian, 4 colon, 2 Non-Hodgkin's disease, 1 bone and 3 with other malignancies. Age (31.7(CI30.4-33.0) vs. 32.4(CI31.3-33.5)), days of stimulation (10.1(CI9.6-10.6) vs. 10.4(CI9.9-10.7)), FSH doses (1472.2(CI(1342.4-1603.0) vs. 1630.5(CI1504.8-1756.2), oocytes retrieved (11.1(CI9.4-12.9) vs. 12.6(CI11.2-14.0), and mature oocytes (8.2(CI6.8-9.6) vs. 9.7(CI8.5-11.0) were statistically comparable.There were differences in E2 on the hCG day (pg/mL) being lower in the cancer group 803.4 (CI539.7-1067.1) vs. 2229.1(CI1923.6-2534.6). CONCLUSION: Similar outcome regarding the number of mature oocytes is achieved by patients with malignancies when compared with normal IVF patients. Ovarian stimulation with letrozole produces on them lower levels of estradiol at hCG day than other protocols.

Redefining advanced maternal age as an indication for preimplantation genetic screening

In this retrospective study, the utility of preimplantation genetic screening (PGS) in patients with advanced maternal age is evaluated. The patient population consisted of women aged 38-44years and included in a regular IVF programme with or without PGS analysis. Transfer rate, ongoing implantation rate and ongoing pregnancy rate were the main outcome parameters measured. A trend of better ongoing pregnancy rate per oocyte retrieval was observed in patients aged 38 and 39years in the non-PGS group when compared with PGS groups, but better ongoing pregnancy rate per oocyte retrieval was observed in patients 41-44years old in the PGS group. When patients with a low ovarian response accumulated oocytes in several stimulation cycles, clinical outcomes were comparable to those of normal-responder patients. These results show that, although PGS does not benefit patients less than 40years of age, reproductive success increases more than two-fold in patients over 40years, especially in patients with more than six metaphase II oocytes, as a result of a good ovarian response or gamete accumulation, suggesting a redefinition of advanced maternal age as indication for PGS. In this retrospective study, the utility of preimplantation genetic screening (PGS) in patients with advanced maternal age is evaluated. Patient population consisted of women aged 38-44 years and included in a regular IVF programme with or without PGS analysis. Transfer rate, ongoing implantation rate and ongoing pregnancy rate were the main outcome parameters measured. A trend of better ongoing pregnancy rate per ovarian stimulation cycle was observed in patients aged 38-39 years in the non-PGS group when compared with PGS groups, but better ongoing implantation rate was observed in patients aged 41-44 years old in the PGS group. When patients with a low ovarian response (low number of oocytes available for the IVF cycle) accumulated oocytes in several stimulation cycles, their reproductive possibilities were comparable to those of normal-responder patients. These results show that, although PGS does not benefit patients less than 40 years of age, reproductive success increases more than 2-fold in patients over 40 years, especially in patients with more than six metaphase II oocytes, as a result of a good ovarian response or gamete accumulation, suggesting a redefinition of advanced maternal age as indication for PGS.

Cumulative reproductive outcome after preimplantation genetic diagnosis: a report on 1498 couples

Couples undergoing preimplantation genetic diagnosis (PGD) have a different background and set of treatment characteristics to couples undergoing regular IVF or ICSI. The aim of this study was to analyse the cumulative reproductive outcome of a large cohort of couples undergoing PGD in relation to a number of explanatory variables potentially affecting the prognosis. Prospective cohort study, Kaplan-Meier analysis was performed to estimate real (observed) and expected (calculated) cumulative delivery rates, and Cox proportional hazard regression analysis was used to assess the effect of age, number of cumulus oocyte complexes collected, fertility status, parity, genetic category and method of pituitary suppression. Between 1993 and 2005, 2753 unselected consecutive cycles of ICSI and PGD were carried out in 1498 couples. The cumulative observed delivery rate overall per couple with a maximum of six treatment cycles of ICSI and PGD performed was 29%. The expected cumulative delivery rate (max six cycles) overall was 62%. There were no significant differences in cumulative delivery rates between the different genetic categories (i.e. availability of transferable embryos after PGD of 50 or 75%, chromosomal translocations or aneuploidy screening). The cumulative reproductive outcome in this PGD cohort was also not significantly affected by the fertility status of the couple, their parity or the method of pituitary suppression. However, the age of the patient and the number of oocytes contributed significantly to the reproductive results. This prospective observational study demonstrates that age has a significantly negative effect on outcome of PGD, due to poor reproductive performance of female partners 40 years of age and older. The number of oocytes collected has a significant and independent effect. The other factors studied did not affect the cumulative reproductive outcome in this PGD cohort.

The impact of oocyte vitrification on laboratory and clinical outcomes in 30 to 38 years old IVF patients

OBJECTIVE: Cryopreservation is known to have significant effects on oocyte physiology, which may compromise the viability of resulting embryos. Vitrification has been successfully applied to human oocyte cryopreservation recently. However, the impact of oocyte vitrification has not been evaluated in the regular IVF patient population. Thus, our objective was to investigate the impact of oocyte vitrification on laboratory and clinical results in typical IVF patients aged between 30 and 38 years.

In-vivo vs. rescued in-vitro maturation metaphase II oocyte vitrification outcomes in human

To achieve better cryopreservation outcome, in-vivo matured oocytes were often chosen as a major source for egg freezing. However, in regular human IVF cycle, immature eggs may comprise 15–20% of total retrieved oocytes. There is little data regarding the survival and developmental potential of those rescued in-vitro maturation (IVM) oocytes after thawing. Therefore, the objective of this study was to evaluate oocyte cryopreservation results comparing in-vivo matured vs. rescued IVM using the vitrification method. Prospective study comparing in-vivo matured vs. in-vitro matured oocyte freeze-thaw outcomes. Oocytes were donated for the study by individuals (<35 y). Two hours after oocyte retrieval, the in-vivo matured oocytes were subject to vitrification. The immature oocytes were cultured to 36 h until PB extrusion, and the rescued IVM oocytes were also frozen by vitrification. The oocytes were first equilibrated in 7.5% Ethylene Glycol (EG) and 7.5% DMSO for 5 min and transferred into the vitrification solution containing 15% EG and 15% DMSO for 1 min before freezing. Oocyte thawing was performed by serial dilutions as follows, 1.0 M, 0.5 M, and 0 M sucrose. Oocytes that survived were inseminated by ICSI 2–3 h after thawing. Fisher's exact test, P<.05. A total 86 in-vivo matured and 63 rescued IVM oocytes were frozen. 74 in-vivo matured oocytes survived (86%), and 51 out of 63 rescued IVM eggs survived (81%; NS) post thawing. Fertilization rate was 92% (68/74) in the in-vivo and 76% (39/51) in the rescued IVM group (P<.005). Cleavage rate was 98% (67/68) in the in-vivo group and 89% (35/39) in the rescued IVM group (NS). 48 out of 68 embryos (71%) developed to blastocyst by day-6 in the in-vivo group, and 6 out of 39 embryos (15%) were observed in the rescued IVM group (P<.0001). Embryo transfer was performed only from the in-vivo matured group. 11 embryo transfers were performed with average 2.5 embryos per transfer, and 10 pregnancies were achieved (91%). A total of 15 FCA were detected in 8 recipients (implantation: 15/21, 71%). The present study demonstrates that both in-vivo and rescued IVM oocytes provide high survival rate after thawing; but fertilization rate and embryo development potential from in-vivo matured group were significantly higher than the rescued IVM group. However, the rescued IVM oocytes may serve as an adjunct to extend the pool of total cryopreserved oocytes.

Gonadotrophin stimulation of non-luteinized granulosa cells increases steroid production and the expression of enzymes involved in estrogen and progesterone synthesis

In regular IVF treatment, mature oocytes are collected with their luteinized granulosa cells (GCs). When in vitro maturation (IVM) of the oocytes is performed, non-luteinized GCs can be collected. We have investigated how these cells respond to gonadotrophin stimulation in culture. GCs were collected from patients undergoing IVM treatment and compared with GCs from IVF patients. The cells were stimulated with FSH and/or hCG. After 48 h, culture media were collected for hormone analysis, and RNA was isolated for gene expression analysis. In IVM GCs, hCG and FSH alone and in combination induced significantly increased progesterone production, and FSH alone and in combination with hCG increased estrogen production. We also studied the gene expression of P-450aromatase and P-450scc and the receptors for FSH and LH. In non-luteinized GCs, the expression levels of P-450aromatase increased with all treatments, and P-450scc expression increased with the combined FSH and hCG treatment. LHR expression increased with FSH treatment, but the FSH receptor expression did not change with different treatments. Non-luteinized GCs behaved differently from luteinized GCs in culture. The data help understand the final stages of maturation of human oocytes and follicles.

P-129: Leuprolide acetate 0.2 cc with human chorionic gonadotropin to trigger ovulation as an effective method to decrease ovarian hyperstimulation syndrome pain without affecting the implantation rate in donor oocyte cycles

Ovarian Hyperstimulation (OHSS) is a significant complication of gonadotropin therapy. Ovulation induction cycles should be highly individualized and should use the minimum effective dosage of Human Chorionic Gonadotropin (HCG) to significantly reduce the risk of OHSS. However, it is unclear if pregnancy rates are compromised by Leuprolide Acetate (LA) trigger alone. Oocyte donors with OHSS will frequently have to take time off from work and will refuse to do another Donor Egg/IVF cycle. The use of LA alone has been shown in several studies to trigger ovulation through an endogenous LH surge. This effect is more physiological than an exogenous HCG trigger, thus reducing the risk of pain and OHSS. This is a retrospective study of donor oocyte patients: Leuprolide Acetate (LA) 1 mg (o.2cc) subcutaneously was given synchronously with 1.25 - 5K HCG. Patients were evaluated for OHSS symptoms (pain 1 to 2 days after the oocyte retrieval), number of oocytes retrieved, peak estradiol levels, number of cryopreserved embryos, and implantation rate. Retrospective review of 33 donor oocyte retrievals from 1/2004 - 1/2006 were evaluated in patients who received LA 0.2cc trigger with HCG. These patients were categorized according to if they had pain versus no pain 1 to 2 days after the retrieval. Ages of the egg donors ranged from 20 to 34. Patients were selected to receive HCG plus LA if their estradiol level ranged from 1600-8800 and the number of follicles greater than 15 ranged from 5-12 A total of 33 donor oocyte patients received LA 0.2cc with HCG. There were 8 shared cycles for a total of 39 transfers. Patients who received LA 0.2cc with HCG had estradiol peak levels ranging from 1618 to 8806, total number of oocytes ranging from 13-68, implantation occurred in 29 recipients (16 singletons, 11 twins, and 2 triplets) with cryopreserved embryos ranging form 0-45. There were a total of 7 patients with complaints of mild to moderate pain and 2 OHSS patients. There was a low incidence of pain in the LA plus HCG donors even though their estradiol levels were high and high number of oocytes retrieved. Oocyte donor patients who receive LA with HCG may have a reduced risk of OHSS symptoms, however the amounts of HCG may be the significant factor. It is prudent for all oocyte donor patients to be assessed for OHSS. Lowering HCG dose and/or using LA may decrease the incidenceof OHSS in a donor oocyte program. Oocytes donors may have less morbidity and may be more likely to pursue additional donation cycles if the incidence of OHSS were lower. Larger studies are needed to compare LA with HCG with donor oocyte patients, as well as regular IVF patients. Other factors to consider are evaluating whether it is the effect of the Gonadotropin Releasing Hormone (GnRH) agonist, LA 0.2cc with HCG or is it the lower dosage of HCG that decreases the incidence of OHSS. Also another question to explore would be if lower doses of HCG would be as effective without LA?

Neonatal and obstetric outcome of pregnancies conceived by ICSI or IVF

Intracytoplasmic sperm injection (ICSI) is currently widely used despite concern regarding pregnancy complications and outcome, specifically congenital malformations. The aim of this study was to compare the obstetric and neonatal outcome of pregnancies conceived by IVF and ICSI. Long-term follow-up was achieved through questionnaires sent to women who conceived after IVF/ICSI treatment. Information was obtained regarding 219 pregnancies (322 children) conceived after ICSI and 145 pregnancies (201 children) conceived after IVF. There were no significant differences between the ICSI and regular IVF pregnancies in regard to the couple's characteristics and the obstetric complications. The mean ± SD birth weight of the singletons conceived after ICSI was similar to that of singletons conceived after IVF: 3001 ± 703 versus 3059 ± 643 g respectively. In both groups there was a high incidence of multiple pregnancies, Caesarean sections, prematurity and low birth weights. The incidence of congenital malformations following IVF or ICSI pregnancies was similar, 6.3 and 7.7% respectively, but was increased compared with the general Israeli population. In summary, concern remains regarding the incidence of congenital malformations after IVF and the long-term outcome of ICSI pregnancies. However, the present results are reassuring with regard to the obstetric and neonatal outcome of pregnancies conceived by ICSI compared with those achieved by IVF.