Improve Embryo Quality Research Articles

Some aspects of interactivity between endocrine and immune systems required for successful reproduction.

BackgroundIn successful reproduction, endocrine and immune systems closely interact. We here attempt to further elucidate the relationship between androgen levels, systemic activation of the immune system and reproductive success in infertile women, utilizing 2 distinct infertile patient cohorts.MethodsIn Group 1, we investigated 322 women (ages 38.6 +/− 5.4 years) at initial presentation; in Group 2 125 women undergoing in vitro fertilization (169 IVF cycles, ages 38.9 +/− 5.5 years). In Group 1, we assessed androgens and an immune panel, previously demonstrated to discriminate between activated quiescent immune systems; in Group 2, utilizing the same immune panel, we investigated whether immune system activation relates to embryo quality in IVF cycles.ResultsNo individual immune test within the immune panel was associated with androgen levels. The total/free testosterone ratio (TT/FT) was, however, significantly associated with presence of gammopathies (in IgG, IgM, IgA, IgE; P = 0.026). Surprisingly, immune system activation was associated with significantly improved embryo quality (P = 0.008), a finding persistent after adjustment for age and repeat IVF cycles (P = 0.006).ConclusionsAssociation of immune system activation with improved embryo quality concurs with previously reported immune activation in association with normal functional ovarian reserve (FOR) and normal androgen levels, while, counter intuitively, hypoandrogenism and low FOR are associated with lack of immune system activation. Mild immune system activation, therefore, likely appears essential for establishment of pregnancy, and may be regulated by androgens.

Fibroblast growth factor 17 and bone morphogenetic protein 15 enhance cumulus expansion and improve quality of in vitro–produced embryos in cattle

Bone morphogenetic protein 15 (BMP15) and members of the fibroblast growth factor (FGF) family are expressed by the oocyte and are involved in the control of cumulus cell function. We tested the hypothesis that FGF17, alone or combined with BMP15 in the maturation medium, enhances cumulus expansion, meiosis progression, embryonic development, and expression of mRNA encoding key genes regulating expansion (prostaglandin-endoperoxide synthase 2 [PTGS2], hyaluronan synthase 2 [HAS2], tumor necrosis factor–stimulated gene 6 [TNFAIP6], and pentraxin 3 [PTX3]) and markers of oocyte developmental competence (phosphofructokinase [PFKP], gremlin [GREM1], versican [VCAN], and the genomic progesterone receptor [nPR]) in cumulus cells. Fibroblast growth factor 17 and BMP15 increased the percentage of fully expanded cumulus–oocyte complexes (COCs), but there was no additive effect when both were combined. Neither FGF17 nor BMP15 altered the percentage of oocytes reaching meiosis II at the end of COC culture or cleavage and blastocyst rates after IVF. However, embryo quality, as assessed by the number of cells in the inner cell mass, was improved by the combination of FGF17 with BMP15. Fibroblast growth factor 17 alone did not alter gene expression in cumulus cells at the end of IVM, whereas BMP15 increased PTGS2 and PTX3 mRNA levels. The combination of FGF17 and BMP15 increased nPR mRNA abundance in cumulus cells but did not change the expression of other markers of developmental competence. This study provides novel evidence that FGF17 enhances cumulus expansion in bovine COCs submitted to IVM and that the supplementation of the IVM medium with FGF17 and BMP15 may improve embryo quality.

The AUGMENTSM Treatment: Physician Reported Outcomes of the Initial Global Patient Experience

Background: Egg precursor cells can be readily isolated from the protective outer lining of the ovarian cortex. Studies have demonstrated that mitochondria isolated from these cells are of high quality. The AUGMENTSM treatment is a proprietary new technology based on previous clinical use demonstrating that the addition of mitochondria during in vitro fertilization (IVF) is safe, improves embryo quality, and increases the success of IVF. Subsequent published animal studies also confirmed the role of mitochondria in improved outcomes. This report represents the earliest global observations of the AUGMENTSM treatment in routine clinical practice from two distinct international centers. Methods and results: The AUGMENTSM treatment was initially used in a population of difficult-to-treat patients with a poor prognosis for success with standard IVF who were likely moving to donor egg as a next step. Each group reported marked improvements in pregnancy rates above the historic IVF success rate for these patients (e.g., 11- and 18- fold increase in ongoing clinical pregnancy rates in the UAE and Canada, respectively). In a 25 patient subset, retrieved eggs from each woman were allocated to two treatment groups; one group underwent the AUGMENTSM treatment at the time of ICSI while the other group underwent conventional ICSI. Embryos were selected for fresh-embryo transfer based on standard criteria including embryo morphology and the results of preimplantation genetic testing. Morphogenetic embryo selection and transfer from the AUGMENTSM treatment group was significantly higher, suggesting that improved embryo quality may have resulted in the improved pregnancy rates observed in these women. Conclusions: Based on these findings, the AUGMENTSM treatment may be a viable treatment option to address an unmet need for women with poor reproductive histories, history of poor egg quality, poor embryo development, previously failed IVF, and those seeking—despite repeated failures-to conceive genetically-related offspring.

Supplementation of bovine embryo culture medium with L-arginine improves embryo quality via nitric oxide production.

Nitric oxide (NO) is a cell-signaling molecule that regulates a variety of molecular pathways. We investigated the role of NO during preimplantation embryonic development by blocking its production with an inhibitor or supplementing in vitro bovine embryo cultures with its natural precursor, L-arginine, over different periods. Endpoints evaluated included blastocyst rates, development kinetics, and embryo quality. Supplementation with the NO synthase inhibitor N-Nitro-L-arginine-methyl ester (L-NAME) from Days 1 to 8 of culture decreased blastocyst (P < 0.05) and hatching (P < 0.05) rates. When added from Days 1 to 8, 50 mM L-arginine decreased blastocyst rates (P < 0.001); in contrast, when added from Days 5 to 8, 1 mM L-arginine improved embryo hatching rates (P < 0.05) and quality (P < 0.05) as well as increased POU5F1 gene expression (P < 0.05) as compared to the untreated control. Moreover, NO levels in the medium during this culture period positively correlated with the increased embryo hatching rates and quality (P < 0.05). These data suggest exerts its positive effects during the transition from morula to blastocyst stage, and that supplementing the embryo culture medium with L-arginine favors preimplantation development of bovine embryos.

Evidence that a degree of mucosal inflammation is beneficial for pregnancy success in association with IVF

We elsewhere in this meeting report that antiphospholipid antibodies (APAs), among a variety of immune parameters, are most closely associated with the diagnosis of low functional ovarian reserve (LFOR). We recently (ASRM 2013) also reported that a degree of immune system activation/inflammation appears associated with improved embryo quality in association with in vitro fertilization (IVF). Attempting to link these observations, we here investigated the impact of immune/inflammatory markers on IVF outcomes.

Undisturbed embryo culture in a time-lapse monitoring system improve embryo quality: a prospective cohort study

Undisturbed embryo culture in a time-lapse monitoring system improve embryo quality: a prospective cohort study

ART and uterine pathology: how relevant is the maternal side for implantation?

Assisted reproduction technology (ART) has become a standard treatment for infertile couples. Increased success rates obtained over the years have resulted primarily from improved embryo quality, but implantation rates still remain lower than expected. The uterus, an important player in implantation, has been frequently neglected. While a number of uterine pathologies have been associated with decreased natural fertility, less information exists regarding the impact of these pathologies in ART. This report reviews the evidence to help clinicians advise ART patients. An electronic search of PubMed and EMBASE was performed to identify articles in the English, French or Spanish language published until May 2014 which addressed uterine pathology and ART. Data from natural conception were used only in the absence of data from ART. Studies were classified in decreasing categories: RCTs, prospective controlled trials, prospective non-controlled trials, retrospective studies and experimental studies. Studies included in lower categories were only used if insufficient evidence was available. Pooled data were obtained from systematic reviews with meta-analyses when available. The summary of the evidence for the different outcomes and the degree of the recommendation for interventions were based on the GRADE (Grading of Recommendations Assessment, Development and Evaluation) statement recommendations. There is strong evidence that surrogacy is effective for uterine agenesia. For the remaining pathologies, however, there is very little evidence that the established treatments improve outcomes, or that these pathologies have a negative effect on ART. In the presence of an apparently normal uterus, assessing endometrial receptivity (ER) is the goal; however diagnostic tests are still under development. The real effect of different uterine/endometrial integrity pathologies on ART is not known. Moreover, currently proposed treatments are not based on solid evidence, and little can be done to assess ER in normal or abnormal conditions. No strong recommendations can be given based on the published experience, bringing an urgent need for well-designed studies. In this context, we propose algorithms to study the uterus in ART.

Effect of leptin on in vivo goat embryo production.

The aim of this study was to evaluate the effect of leptin administration during superovulation on in vivo goat embryo production. Ten mature does were superovulated with 133mg follicle-stimulating hormone (FSH) i.m. in six descending doses at 12-h intervals. The goats received 4.8μg/kg human recombinant leptin s.c. (leptin group, n=5) or phosphate-buffered saline (PBS) (control group, n=5) with the first and second FSH doses. The does were mated and subjected to embryo collection by transcervical technique 6days later. The total number of cells per embryo and the number of cells with fragmented DNA were assessed in selected blastocysts by combining Hoechst 33342 and terminal dUTP nick-end labelling (TUNEL) staining. Plasma concentrations of oestradiol (E2 ) and progesterone (P4 ) were determined by electrochemiluminescence from the day of FSH treatment, on the day of superovulatory oestrus and on the day before embryo collection. Compared with the control group, the does that received leptin had a higher number of transferable embryos (p<0.005), fewer embryos classified as degenerated (p<0.001) and fewer TUNEL-positive cells/blastocyst (p<0.001). The number of transferable embryos was positively correlated with E2 concentrations on day of oestrus (r=0.562; p<0.01) and P4 concentrations on the day of embryo collection (r=0.912; p<0.001). We concluded that in vivo leptin administration during FSH treatment improved embryo quality and affected ovarian steroidogenesis in superovulated goats.

Differential expression of selected candidate genes in bovine embryos produced in vitro and cultured with chemicals modulating lipid metabolism

Lipid accumulated in embryos produced in vitro has been linked to reductions in both quality and postcryopreservation viability. Therefore, the objective of the present study was to investigate the influence of lipid-reducing chemicals on embryo development, quality, and postcryopreservation viability, in addition to expression profiles of selected lipid metabolism-regulating genes. Bovine cumulus-oocyte complexes were matured and fertilized in vitro; eight-cell stage embryos were cultured in IVC medium supplemented with phenazine ethosulfate (PES), l-carnitine (LC), PES + LC, or no supplementation (control). Culturing embryos in medium with LC increased (P < 0.05) blastocyst rate (38.8%) compared with the other groups (control = 28.1%, PES = 27.1%, PES + LC = 26.3%). Embryos cultured with supplements had greater total cell number and fewer apoptotic cells than the control. Cytoplasmic lipid content was reduced, whereas mitochondria density was increased in embryos treated with culture supplements; this was linked to altered expression profiles of selected genes regulating lipid metabolism. For example, transcript abundance of transmembrane lipid gene (SGPP1) was greater in LC- and PES-treated embryos, and they had increased postcryopreservation hatching ability (indicative of embryo cryotolerance). In conclusion, the two lipid metabolism regulators added to the culture media had improved embryo quality and cryotolerance, but embryo development rate and downstream lipid metabolism-regulating genes were more influenced with LC supplementation.

Effects of sorbitol on porcine oocyte maturation and embryo development in vitro

In the present study, a porcine system was supplemented with sorbitol during in vitro maturation (IVM) or in vitro culture (IVC), and the effects of sorbitol on oocyte maturation and embryonic development following parthenogenetic activation were assessed. Porcine immature oocytes were treated with different concentrations of sorbitol during IVM, and the resultant metaphase II stage oocytes were activated and cultured in porcine zygote medium-3 (PZM-3) for 7 days. No significant difference was observed in cumulus expansion and the nuclear maturation between the control and sorbitol-treated groups, with the exception of the 100 mM group, which showed significantly decreased nuclear maturation and cumulus expansion. There was no significant difference in the intracellular reactive oxygen species (ROS) levels between oocytes matured with 10 or 20 mM sorbitol and control groups, but 50 and 100 mM groups had significantly higher ROS levels than other groups. The 20 mM group showed significant increases in intracellular glutathione and subsequent blastocyst formation rates following parthenogenetic activation compared with the other groups. During IVC, supplementation with sorbitol significantly reduced blastocyst formation and increased the apoptotic index compared with the control. The apoptotic index of blastocysts from the sorbitol-treated group for entire culture period was significantly higher than those of the partially sorbitol-exposed groups. Based on these findings, it can be concluded that the addition of a low concentration of sorbitol (20 mM) during IVM of porcine oocytes benefits subsequent blastocyst development and improves embryo quality, whereas sorbitol supplement during IVC has a negative effect on blastocyst formation.

32 PREGNANCY OF EQUINE CLONED EMBRYOS MICROINJECTED WITH PLURIPOTENCY INDUCING GENES (Oct4, Sox2, c-Myc, K1f4)

Animal cloning is a high impact tool for scientific and economical production, but still with inefficient results. The efficiency of the cloning process depends on the state of differentiation of the donor cell. An adult equine somatic cell can be differentiated to a pluripotent stem cell (iPSC) inducing the expression of certain transcription factors (Oct4, Sox2, c-Myc, and K1f4; Breton et al. 2013). The objective of this work was to assess the effect of the intracytoplasmic injection of pluripotency inducing genes on embryo development and pregnancy rates of equine cloned embryos. Cumulus–oocyte complexes (COC) were obtained from slaughterhouse ovaries. Oocyte collection and maturation procedure were performed as described by Lagutina et al. (2007). After the removal of cumulus cells, oocytes showing first polar body were microinjected with a mixture 1/3 of plasmids/liposomes (Mi group). The plasmid used was the pEP4-E02s-EM2k, which encodes the human genes Oct4, Sox2, Myc, and K1f4. The DNA concentration was adjusted to 0.5 μg mL–1. Microinjected oocytes were enucleated using the zona free method. Adult male skin fibroblasts from the same animal were used as donor nucleus cells. These fibroblasts were attached to the ooplasts with phytohemagglutinin and then fused with an electric pulse. Activation was performed using 8.7 mM ionomycin for 4 min, followed by culture for 4 h in a combination of 1 mM 6-DMAP and 5 mg mL–1 cycloheximide. Zona free reconstructed embryos (ZFRE) were cultured for 7 to 8 days in DMEM-F12 in the well of the well (WOW) system, aggregating 3 embryos per well. A control group (CC group) of not microinjected embryos was included. Cleavage and blastocyst development was assessed at Days 2 and 7, respectively. Transcervical transfer of 49 Day 7 to 8 blastocysts was performed 6 days after ovulation. The mares received 2 blastocysts per transfer. Pregnancy was diagnosed by transrectal ultrasonography 15 days after ovulation. Cleavage and blastocyst rates were analysed by Chi-squared test and pregnancy rate by Fisher test (P &lt; 0.05). Cleavage was 92.1% (n = 58/63) for the Mi group and 90.4% (n = 868/960) for the CC group. Blastocyst rate was statistically higher per well, 28.6% (n = 6/21) v. 13.4% (n = 43/320) but not per oocyte, 9.5% (n = 6/63) v. 4.5% (n = 43/960), for the Mi and CC groups, respectively. Pregnancy rate was 17% (n = 1/6) for the Mi group and 7% (n = 3/43) for the CC group. No twin pregnancies were found and all the pregnancies are still ongoing. The higher blastocyst rates obtained with the embryos microinjected with pluripotency inducing genes compared with the control group showed an improvement in embryo quality. In conclusion, the data presented indicate that the intracytoplasmic microinjection of pluripotency inducing genes in equine zona free cloned embryos improved blastocyst rates on a per well basis and showed a tendency to improve the pregnancy rates. The expression of the Oct4, Sox2, c-Myc, and K1f4 genes could be probably generating better reprogrammed donor nucleus compared with adult differentiated cells used in conventional cloning.

84 THE EFFECT OF β-MERCAPTOETHANOL ON CLEAVAGE RATES, DEVELOPMENTAL COMPETENCE AND QUALITY OF IN VITRO PRODUCED BOVINE EMBRYOS

The production of excessive levels of reactive oxygen species can be a major problem during in vitro embryo culture. Although studies have shown that supplementation with exogenous antioxidants can improve embryo quality, the results are controversial among researchers. In this study, we examined the effects of different concentrations of β-mercaptoethanol (β-ME) added to the culture media, on cleavage rates, the quality and developmental competence of in vitro-produced bovine embryos. The embryos were produced in vitro as described previously (Van Hoeck et al., 2013). Briefly, in total, 753 grade I cumulus–oocyte complexes (COC) from 2- to 6-mm-diameter follicles were matured in groups of 50 in 500 μL of TCM with 20 ng mL–1 EGF for 24 h, fertilized in groups of 100 in 500 μL of fertilization medium for 20 h (5% CO2, 38.5°C). Presumptive zygotes were denuded and randomly assigned to 4 treatments with different concentrations of β-ME: 0 μM (control), 50 μM, 100 μM, and 150 μM. They were cultured in groups of ±25 in 50 μL of SOF supplemented with ITS (10 μg mL–1 insulin; 5.5 μg mL–1 transferrin; 6.7 ng mL–1 selenium) and 2% BSA and covered with mineral oil (5% O2, 5% CO2, 38.5°C). At 48 h post-insemination (p.i.), cleavage rate was evaluated and expressed as the number of cleaved embryos on total number of oocytes. At Day 7 p.i., blastocyst rate was determined (number of blastocysts on total number of oocytes), blastocysts were fixed in 4% paraformaldehyde, and total cell number was determined by DAPI staining. Data were analysed by ANOVA and post hoc test. Comparable cleavage rates were obtained in treated groups: control (80.8%), 50 μM (77.7%), 100 μM (77.9%), and 150 μM (73.6%; P &gt; 0.05). Also, no significant effect of treatment could be found on blastocyst rates: control (36%), 50 μM (36.5%), 100 μM (38.4%), and 150 μM (30.4%). The total cell number per blastocyst increased significantly (P &lt; 0.05) using 100 μM of β-ME compared with the controls (158.0 ± 24.3 v. 123.2 ± 9.72, respectively). These results suggest that the inclusion of 100 μM β-ME during in vitro embryo culture could be used for production of high quality bovine blastocysts.

Twice-daily dosing of gonadotropins does not improve embryo quality during in vitro fertilization cycles in women with polycystic ovary syndrome, when compared to once-daily dosing: a pilot study

To determine whether overexpression of the FSH receptor in polycystic ovary syndrome (PCOS) results in a relative deficiency of gonadotropins and poor oocyte and embryo quality during in vitro fertilization (IVF) cycles. Whether twice-daily dosing of gonadotropins could therefore result in improved embryo quality, by fixing this hypothesized relative deficiency of gonadotropins. Embryos generated at a university-based fertility center in women with PCOS were compared from twice-daily dosing to once-daily dosing of gonadotropins during IVF cycles. Oocyte and embryo quality was compared. A single patient's embryos were included in the analysis from only one IVF cycle and all embryos from that cycle were included. 254 embryos were compared. Twice-daily vs. once-daily dosing of gonadotropins does not improve embryo or oocyte quality in women with PCOS. The defect in response to gonadotropins in PCOS is most likely due to an inherent defect in the ovary and not a relative deficiency of gonadotropins due to overexpression of the FSH receptors. More studies are needed to confirm this finding.

Intracytoplasmic morphologically selected sperm injection is beneficial in cases of advanced maternal age: a prospective randomized study

ObjectiveTo evaluate advanced maternal age as a rationale for performing intracytoplasmic morphologically selected sperm injection (IMSI). Study designThis study included couples undergoing intracytoplasmic sperm injection (ICSI) as a result of advanced maternal age (≥37 years old). Sample size calculations were based on the assumption that a 15% difference in implantation rate would mean a clinically significant difference. To achieve this difference, 33 cycles would be needed in each treatment arm (with a significance level of 5% and power of 85%). Couples were randomly allocated to one of two sperm selection procedures (ICSI, n=33; or IMSI, n=33). Sperm selection in the ICSI group was analyzed under a magnification of 400×. Sperm selection in the IMSI group was analyzed under high magnification of 6600×. The groups were compared with regard to the outcome of the cycles. ResultsIMSI cycles showed significantly higher implantation (4/33, 12.1% vs. 18/47, 38.3%, p=0.026) and pregnancy (4/29, 13.8 vs. 18/30, 60.0%, p<0.001) rates. The IMSI procedure positively influenced the blastocyst formation rate (RC: 15.00, R2: 49.9%, p=0.001) and implantation rate (RC: 24.04, R2: 9.6, p=0.027), and was determinant to the increased odds of pregnancy (OR: 9.0, CI: 2.17–37.38, p=0.001). ConclusionIt seems that the injection of a morphologically normal spermatozoon overcomes the low oocyte quality in older women, resulting in improved embryo quality and in a 9-fold increase in the clinical pregnancy rate in couples with advanced maternal age.

Is immune system activation/systemic inflammation a prerequesite for successful reproduction? association of immune system activation with embryo quality during in vitro fertilization (IVF)

Requiring implantation and tolerance of semi-allograft, pregnancy mandates complex immunological processes. Degree of immune system activation has been associated with pregnancy complications. Effects of immune system activation on embryo quality has, however, never been investigated. Cohort study. 125 infertile women underwent 169 IVF cycles. Embryo quality was defined as good (4-cell d-2; 8-cell d-3, Grade 4-5), poor (arrested or degenerated, Grade ≤ 2) or fair (all embryos in between). Immune system activation was tested utilizing immune parameters, demonstrated effective in prior studies in identifying immune system activation with good sensitivity: thyroid antibodies (peroxidase, thyroglobulin, thyroid receptor antibodies), total immunoglobulins (IgG, IgM, IgA, IgE), antiphospholipid antibodies (β2 glycoprotein, anticardiolipin, antiphosphatidylserine, lupus anticoagulant), antinuclear antibody, anti-adrenal antibody (21-hydroxylase) and anti-ovarian antibodies. Any positive finding was considered indicative of immune activation. Patient age was 38.9 ± 5.5 years. Immune system activation was associated with significantly better embryo quality (P=0.008). This association persisted after adjustment for age and repeat IVF cycles (P=0.006). Observed improvement in embryo quality derived mostly from better embryo grade (3.7 ± 0.05 vs. 3.4 ± 0.07, P=0.002). This is the first report associating improved embryo quality with maternal immune system activation. These data relate to recent report from our center that normal androgen levels appear associated with immune system activation, while low androgen levels fail to show such activation (Gleicher et al ESHRE 2013/Reprod Biol Encocrinol 2013; In press). Combined, these data suggest that immune system activation/systemic inflammation, possibly via androgen effects on follicle maturation, contributes to embryo quality and pregnancy success with IVF.

Three day pre-treatment with GnRH antagonists before controlled ovarian stimulation in IVF/ICSI: efficacy from clinical and embryological perspective

The aim of this study was to investigate whether three-day administration of GnRH antagonists in the early follicular phase prior to the start of controlled ovarian stimulation (COS) with rFSH in IVF/ICSI cycles improves embryo quality and clinical pregnancy rate in young patients with good ovarian reserve, compared to standard GnRHant protocol.

The comparison of the effect on IVF results of metformin and insulin used during IVF cycles of infertile women with overt diabetes

To compare the effect of metformin and insulin on controlled ovarian stimulation (COS) and IVF results in infertile women with type 2 diabetes undergoing IVF/intracytoplasmic sperm injection (ICSI). Retrospective cohort study. A total of 52 consecutive IVF/ICSI cycles were included in 35 women with preexisting type 2 diabetes who were treated with either the metformin (metformin group, 29 cycles) or the insulin (insulin group, 23 cycles) for glycemic control. IVF/ICSI cycles in which a GnRH antagonist multiple dose protocol (MDP) is used for COS was included in this study. There were no significant differences in patient's characteristics between metformin and insulin groups. There were also no differences in total dose and days of recombinant human FSH (rhFSH) administered, and numbers of retrieved oocytes, mature oocytes, fertilized oocytes, and embryos transferred between the two groups. However, the number of grade 1 or 2 embryos was significantly higher in metformin group of 3.9 ± 2.9 compared with 2.2 ± 2.3 in insulin group (P = .025). Follicular fluid (FF) TNF-a and IL-6 concentrations at oocyte retrieval were significantly lower in metformin group than in insulin group (P = .030, P = .009). There were no differences in the clinical pregnancy rate, embryo implantation rate, miscarriage rate and multiple pregnancy rate between the two groups. The use of metformin for glycemic control is more effective in reducing FF TNF-a and IL-6 levels in COS cycles and improving embryo quality compared with insulin treatment in infertile women with type 2 diabetes undergoing IVF/ICSI.

Ovarian stimulation using low-dose menotropins in combination with clomiphene citrate results in better embryo quality than gonadotropin-only protocols

Patients undergoing IVF that result in poor embryo quality have a decreased chance of success. Most ovarian stimulation protocols use only gonadotropins dosed according to ovarian reserve. We studied whether a combination of clomiphene citrate and low-dose menotropins improves embryo quality in patients who had a history of poor embryos in gonadotropin-only cycles.

Importance of the GDF9 signaling pathway on cumulus cell expansion and oocyte competency in sheep

Acquisition of developmental competency in cumulus oocyte complexes (COCs) is derived from endocrine hormones and oocyte secreted factors. The contribution of these factors in oocyte maturation and development is an active area of research. The objective of this research was to investigate whether growth differentiation factor 9 (GDF9) that is secreted by oocyte affects cumulus expansion and oocyte development in sheep. Immature ovine COCs were cultured in the presence of recombinant human GDF9 (rhGDF9), denuded oocytes, SB-431542, a specific inhibitor of activin-like kinase 4/5/7; or a combination of these factors. Routine in vitro maturation of COCs and denuded oocytes were used as external control samples. Cultured COCs were used for assessment of (1) cumulus expansion; (2) expression of cumulus-related transcripts including pentraxin 3, hyaluronan synthase 2 (HAS2), tumor necrosis factor alpha-induced protein 6, prostaglandin synthase 2, B-cell lymphoma 2 (BCL2), and Bcl2-associated X (BAX); and (3) yield and quality of embryo development. It was observed that cumulus expansion was not affected by any of these treatments. HAS2 mRNA expression confirmed this observation. In the presence of exogenous GDF9, cleavage rate was reduced, blastocyst rate did not differ from other groups, and trophectoderm cell number significantly increased. This suggests that exogenous GDF9 could improve embryo quality. It was also observed that oocyte secreted factors reduced proapoptotic BAX mRNA, and BCL2 mRNA expression was not significantly different from other groups. This study provides evidence that GDF9 signaling might have a minor influence on ovine cumulus expansion and oocyte development and that other signaling pathway(s) might have a dominant role.

A comparison of the Cook single lumen immature ovum IVM needle to the Steiner-Tan pseudo double lumen flushing needle for oocyte retrieval for IVM

This study compared the impact of using the Steiner-Tan pseudo double lumen needle for antral follicle oocyte retrieval to using a conventional non-flushing needle. The Steiner-Tan needle has a much smaller dead space than the needles commonly used for IVM oocyte retrievals. This was a retrospective cohort study. The patient population was determined by the time period in which a patient underwent IVM in a single physician's IVF practice. The following data was abstracted from clinical and embryology records: oocytes retrieved, oocytes matured, early maturing oocytes, oocytes fertilized, embryo quality measures, retrieval time, needle punctures, clot formation, and clinical pregnancy rate. The Steiner-Tan needle did not increase the number of oocytes retrieved. It also did not increase the time required for retrieval. However, flushing of antral follicles significantly decreased clot formation in fluid aspirates. Use of the Steiner-Tan needle also significantly decreased the number of vaginal needle punctures during each case. There was a trend toward improved embryo quality, but statistical power was inadequate to show a difference. The primary benefit of the Steiner-Tan needle was on the embryological aspects of IVM. Decreased blood and blood clots in the aspirates made an IVM retrieval more like conventional IVF for the embryologist. The patient also experienced less tissue trauma without increasing anesthesia or surgical time. There was no improvement in the number of oocytes retrieved, but based on the results, we hypothesized that oocytes were more commonly retrieved from slightly large follicles than when using a routine needle.