Improve Embryo Quality Research Articles

Superovulatory response and embryo quality in Boer does following dietary supplementation with different sources of omega-3 and omega-6 fatty acids during the breeding season

The purpose of this study was to determine effects of various sources of omega-3 and omega-6 fatty acids on ovarian response and embryo quality in Boer does when there was a superovulation treatment regimen imposed. Pluriparous does were randomly assigned to be treated with 300 g of one of four experimental supplements containing linseed oil (LO), soybean oil (SO), palm oil (PO), or a control supplement without fatty acids (CO), for 15 days. Does were fitted with a controlled internal drug release (CIDR) device containing 0.3 g progesterone for 7 days. At 48 h before CIDR withdrawal, does were treated with 80 mg follicle-stimulating hormone (FSH) administered at 12 h intervals. Embryos were collected 7 days after the last natural mating. Estrous response and interval between CIDR withdrawals to estrous onset were similar between treatments (P > 0.05). Number of ovulations was similar for does in the different groups (10.0, 9.2, 7.0, and 7.0, in LO, SO, PO, and CO, respectively; P > 0.05). There was premature luteal regression in does of the SO, PO, and CO groups, except in LO group. The LO-treated does had a larger (P < 0.05) mean number of ova/embryos recovered than does of SO, PO, and CO groups (7.2, 2.0, 0.2, 0.2, respectively) and transferable embryos (5.1, 1.4, 0.2, 0.2, respectively). These results indicate that including LO in supplements may be a feasible strategy for preventing premature luteal regression and improving embryo quality in goats treated to induce follicular super-stimulation for induction of superovulation.

Fresh oocyte cycles yield improved embryo quality compared with frozen oocyte cycles in an egg-sharing donation programme.

The objective of this study was to investigate any effect of cryopreservation of donated eggs on laboratorial and clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles. This retrospective cohort study included 320 oocyte recipients undergoing 307 vitrified and 119 fresh oocyte recipient ICSI cycles, participating in an egg-sharing donation programme, from 2015 to 2018, in a private university-affiliated in vitro fertilization (IVF) centre. A review of donor and recipient ICSI cycles was charted. A general mixed models fit by restricted maximum likelihood, followed by Bonferroni post hoc test was used to compare the means between fresh and warm oocyte donation groups and investigate the effect of cryopreservation on recipient ICSI outcome. The main outcome measure was blastocyst development rates. Fertilization rate, high-quality embryo rates on days 2 and 3, normal cleavage speed rates on days 2 and 3, and blastocyst development rate were significantly higher for the fresh oocyte donation cycles compared with warmed oocyte donation cycles. In the egg-sharing donation programme, fertilization and embryo developmental competence were reduced when vitrified oocytes from infertile couples were used for ICSI compared with fresh oocytes.

52 Extracellular vesicles from oviduct and uterus in sequential invitro culture affects mitochondrial activity and lipid metabolism transcripts in bovine embryos

Oviducal fluid (OF) and uterine fluid (UF) improve the quality of embryos during invitro culture, and their extracellular vesicles (EV) may be involved in such an effect. We aimed to evaluate the effect of EV from OF and UF in sequential invitro culture on the development and quality of bovine embryos. Zygotes were cultured in synthetic oviduct fluid supplemented either with 3mg mL−1 BSA (n=1584) or 5% EV-depleted fetal calf serum (dFCS, n=1594) in absence or presence (BSAEV, n=1853 and dFCSEV, n=1473) of 3×105 EV mL−1 from OF (Day 1 to Day 4) and UF (Day 5 to Day 8), mimicking invivo conditions. EV from oviducts (early luteal phase) and uterine horns (mid luteal phase) from slaughtered heifers were isolated by size exclusion chromatography; size and concentration were assessed by nanotracking analysis (NTA) and morphology by transmission electron microscopy (TEM). Blastocyst rate was recorded on Days 7–8 and their quality was assessed for mitochondrial activity by staining with Mitotracker Deep Red (ThermoFisher Scientific), survival rate after vitrification/warming by invitro culture for up to 72h, and relative mRNA abundance of lipid metabolism-related transcripts by quantitative PCR. Housekeeping genes were H2AFZ and ACTB. Data were analysed by one-way ANOVA and Tukey test. TEM confirmed the presence and morphology of EVs, and NTA indicated mode size and concentration of particles (137.2 and 151.2 nm; 2.97×1010 and 7.98×1010 particles mL−1, for OF and UF, respectively). Blastocyst yield was lower (P&amp;lt;0.05) in BSA groups compared with dFCS groups (BSA: 16.2±1.5 and 31.0±1.9; BSAEV: 14.1±1.6 and 26.2±2.0% vs. dFCS: 30.5±2.0 and 40.6±2.4; dFCSEV: 31.1±2.5 and 39.8±2.7%, Day 7 and Day 8, respectively), irrespective of EV supplementation. Blastocyst mitochondrial activity was increased (P&amp;lt;0.05) by EV in dFCSEV compared with the other groups. No differences in survival rate after vitrification/warming were found (range at 72 h: 67.1±8.1 to 87.8±5.7%). PPARGC1B was downregulated and ACC upregulated by EV, irrespective of protein source in medium (P&amp;lt;0.05). In contrast, EV affected some transcripts depending on the protein source in the medium (CD36 upregulated in dFCSEV, downregulated in BSAEV; PLIN2 downregulated in dFCSEV and ATGL downregulated in BSAEV, P&amp;lt;0.05). In conclusion, mimicking physiological conditions using EV from OF and UF in sequential IVC does not affect development but improves embryo quality by increasing blastocysts’ mitochondrial activity and favours the expression of specific lipid metabolism transcripts. Functional effects of EV may be influenced by the protein source in the medium. This research was funded by MINECO-Spain AGL2015-70140-R, PID2019-111641RB-I00, RTI2018-093548-B-I00; YN Cajas, SENESCYT-Ecuador; CLV Leal, FAPESP-Brazil 2017/20339-3, CNPq-Brazil 304276/2018-9.

Dual Trigger with Gonadotropin-releasing Hormone Agonist and Human Chorionic Gonadotropin Improves Live Birth Rate for Women with Expected Normal Ovarian Response in Gonadotropin Releasing Hormone Antagonist Cycles: Retrospective Study

Objective: To evaluate and compare cycle outcomes following triggering final oocyte maturation with dual trigger of concomitant gonadotropin-releasing hormone (GnRH) agonist and human chorionic gonadotropin (hCG) administration versus hCG alone for women with expected normal ovarian response that underwent antagonist cycles with intracytoplasmic sperm injection (ICSI). Material and Methods: Women with expected normal ovarian response that underwent GnRH antagonist cycles with ICSI between January 2010 and April 2020 were evaluated in this retrospective cohort study. A total of 2,443 patients were included. Dual trigger was used for oocyte maturation in 637 cycles whereas hCG alone was used for triggering in 1,806 women. Cycles with dual trigger were assigned to study group and cycles with hCG alone are taken as controls. Results: Number of retrieved oocytes (14.08±3.58 vs. 13.15±3.61), number of metaphase 2 oocytes (9.77±3.08 vs. 8.06±3.14), fertilization rate (0.75±0.19 vs. 0.69±0.19), implantation rate (0.43±0.48 vs. 0.35±0.50) and clinical pregnancy rate (49.9% vs. 40.6%) were significantly higher in dual trigger group in comparison to hCG alone group. Higher number of good quality embryos were obtained in dual trigger group (85.7% vs. 76.3%). Live birth rate was significantly increased in dual trigger group in comparison to hCG only trigger group (45.1% vs. 36.7%). Multivariate logistic regression analysis showed dual trigger is a significant factor in predicting live birth deliveries (odds ratio 1.426, 95% confidence interval 1.185-1.716). Conclusion: Dual-triggering appears to improve embryo quality, increase implantation rates, clinical pregnancy rates and live birth rates in women with expected normal ovarian response that underwent GnRH antagonist cycles.

139 Effect of lycopene supplementation in maturation medium on production of reactive oxygen species and post-vitrification quality of bovine blastocysts

Excessive production and accumulation of reactive oxygen species (ROS) may cause embryo damage associated with oxidative stress. Lycopene, a natural antioxidant, can scavenge singlet oxygen and is one of the most effective antioxidants among carotenoids. We evaluated the effects of supplementation of lycopene (antioxidant), menadione (prooxidant), and their combination during invitro oocyte maturation on ROS generation in matured oocytes and the quality of vitrified-warmed embryos. Cumulus–oocyte complexes, collected from the slaughterhouse, were matured in groups of 60 in 500μL of TCM-199 medium+50mg mL−1 gentamycin+20ng mL−1 epidermal growth factor, for 22h at 38.5°C in 5% CO2 in air and then supplemented with (1) 0.2μM lycopene, (2) 5μM menadione, (3) 0.2μM lycopene+5μM menadione (L+M), or (4) not supplemented (control). Fertilization and embryo culture were performed similarly for all the groups. In the first experiment, ROS measurement (n=236; via fluorescent microscopy) was performed in denuded, matured oocytes incubated in 5μM CellROX® Green (ThermoFisher Scientific) for 1h. Fluorescent intensity was measured in Image-J. In the second experiment, embryos in the blastocyst stage (n=143) were vitrified as previously described by Ortiz-Escribano et al. (2017 Biol. Reprod. 96, 288-301). Vitrified blastocysts were then warmed and washed in decreasing concentrations of sucrose and incubated for 2 days in culture medium [50µL of synthetic oviductal fluid (SOF)+(5g mL−1 insulin, 5g mL−1 transferrin, 5ng mL−1 selenium)]. The quality of vitrified-warmed blastocysts was assessed using a differential staining as described by Wydooghe et al. (2011 Anal. Biochem. 416, 228–230). The effects of pro- and antioxidant supplementation on oocyte fluorescent intensity and embryo quality parameters were fitted in linear mixed-effects models, and results are expressed as least squares means and standard errors. The fluorescent intensity for ROS was lower (P&amp;lt;0.05) in lycopene (10.06±2.92) than in menadione (16.8±2.92). No differences (P&amp;gt;0.05) in ROS intensity values were found among the other groups [control (13.5±2.92) and L+M (13.7±2.90)]. Total cell number (TCN) was similar (P&amp;gt;0.05) in lycopene (153±2.95), L+M (143±4.59), and control (145±3.67) but lower (P&amp;lt;0.05) in menadione (134±6.08). Lesser numbers of apoptotic cells (AC) and AC/TCN values (P&amp;lt;0.05) were recorded in lycopene (4.12±3.07 and 2.71±2.21) compared with control (6.18±3.82 and 4.31±2.75), L+M (6.00±4.79 and 4.22±3.45), and menadione (7.75±6.33 and 5.82±4.56). For the remaining embryo quality parameters, no differences were found (P&amp;gt;0.05). In conclusion, lycopene supplementation during invitro oocyte maturation effectively scavenged free radicals, lowering oxidative stress and improving embryo quality post-vitrification and warming.

Extracellular Vesicles and the Oviduct Function.

In mammals, the oviduct (or the Fallopian tube in humans) can be divided into the infundibulum (responsible for oocyte pick-up), ampulla (site of fertilization), isthmus (where preimplantation embryos develop), and uterotubal junction (where embryos transit to the uterus). The oviductal fluid, as well as extracellular vesicles produced from the oviduct epithelial cells, referred to as oEVs, have been shown to improve the fertilization process, prevent polyspermy, and aid in embryo development. oEVs contain molecular cargos (such as miRNAs, mRNAs, proteins, and lipids) that can be delivered and fuse to recipient cells. oEVs produced from the ampulla appear to be functionally distinct from those produced from the isthmus. In multiple species including mice, cats, dogs, pigs, and cows, oEVs can be incorporated into the oocytes, sperm, and embryos. In this review, we show the positive impact of oEVs on gamete function as well as blastocyst development and how they may improve embryo quality in in vitro conditions in an assisted reproductive technology setting for rodents, domestic animals, farm animals, and humans.

Embryo incubation by time-lapse systems versus conventional incubators in Chinese women with diminished ovarian reserve undergoing IVF/ICSI: a study protocol for a randomised controlled trial

IntroductionThe time-lapse imaging system (TLS) is a newly developed non-invasive embryo assessment system. Compared with conventional incubators, a TLS provides stable culture conditions and consistent observations of embryo development, thereby...

Oocyte Vitrification Temporarily Turns on Oxidation-Reduction Process Genes in Mouse Preimplantation Embryos.

We aim to understand how oocyte vitrification impacts subsequent mouse preimplantation embryo development at molecular level. We profiled transcriptomics of fertilized preimplantation embryos derived from mouse vitrified-warmed oocytes. Concomitantly, we evaluated epigenetic markers in fertilized preimplantation embryos. We found that oocyte vitrification did not affect the fertilization and cleavage process but delayed embryo development until blastocyst stage. RNA sequencing revealed that 1575 genes were profoundly altered in the 2-cell stage embryos developed from vitrified oocytes. The most significantly altered biological pathway was "oxidation-reduction process." Such profound transcriptomics change was associated with decreased level of oocyte-specific histone H1FOO in zygote and 2-cell stage. Transcriptome alteration due to oocyte vitrification was less pronounced as embryos develop into the morula stage. Oocyte vitrification temporarily changes transcriptomics in early preimplantation embryos. Targeting oxidation-reduction pathway might be a potential therapeutic strategy to improve embryo quality and long-term embryo survival.

SUPERIOR SPERM SELECTION? MICROFLUIDIC SPERM SORTING IMPROVES EUPLOID EMBRYO ONGOING PREGNANCY RATE COMPARED TO DENSITY GRADIENT CENTRIFUGATION

Euploid embryo pregnancy failures are frustrating for clinicians and patients alike and remain a poorly understood aspect of infertility. Novel sperm selection techniques using microfluidic sorting devices are reported to improve embryo quality and pregnancy outcomes, though existing data is limited by small sample size. This study aims to compare euploid embryo ongoing pregnancy rates from embryos generated with the use of microfluidic sperm sorting (MSS) versus standard density gradient centrifugation (DGC).

Improvement instead of stability in embryo quality between day 3-5: A possible extra predictor for blastocyst selection

ObjectiveThe aim of this study was to evaluate the predictive value of the dynamic morphological development process between cleavage-stage and blastocyst-stage embryos. Study designA retrospective study was executed between 2015 and 2017 at Ghent University Hospital. A total of 996 first fresh IVF/ICSI cycles resulting in a single embryo transfer on day 5 were included. Embryos were scored on day 3 and day 5 as excellent, good, moderate or poor based on Alpha/ESHRE guidelines and Gardner and Schoolcraft scoring-system. If embryos changed category between day 3 and 5, the number of steps (between excellent; good; moderate; poor) in positive and negative direction was expressed. ResultsOn day 5, the ongoing pregnancy rate (OPR) of excellent embryos was 37.4 %. Univariate analyses showed that on day 5, both a higher cell stage, better inner cell mass and better trophectoderm were significantly associated with an ongoing pregnancy. In case of deterioration in quality of individual embryos between day 3 and day 5, the OPR was significantly lower. Conversely, improvement of embryo quality between day 3 and day 5 showed higher ongoing pregnancy rates (overall OPR of good day-3 embryos improving to excellent day-5 embryos: 30 %; moderate day 3 to excellent day 5: 50 %; poor day 3 to excellent day 5: 42 %; poor day 3 to good day 5: 20 %; poor day 3 to moderate day 5: 16 %). When embryos improved from poor on day 3 to excellent day 5 the OPR was significantly higher in comparison with embryos that did not change in quality scoring during development (steady embryos) (OR: 1.785, p < 0.05). ConclusionOur results suggest that it is more likely to achieve an ongoing pregnancy when transferring an embryo that has improved in quality between days 3 and 5 as opposed to one that has remained stable.

Effect of Dickkopf-1 and colony stimulating factor-2 on the developmental competence, quality, gene expression and live birth rate of buffalo (Bubalus bubalis) embryos produced by hand-made cloning

A functional canonical WNT signaling pathway exists in preimplantation embryos and inhibits embryonic development. Recent studies suggest that this pathway is over-expressed in nuclear transferred (NT), compared to IVF embryos. The present study investigated the effects of Dickkopf-1 (DKK1), an inhibitor of canonical WNT signaling pathway and colony stimulating factor-2 (CSF2), an embryokine, on the developmental competence, quality, gene expression and live birth rate of NT buffalo embryos produced by Hand-made cloning (HMC). Following supplementation of the in vitro culture medium on day 5 with DKK1 (100 ng/mL), CSF2 (10 ng/mL), DKK1+CSF2 or no supplementation (control), the blastocyst rate was higher (P < 0.05) with DKK1 and DKK1+CSF2 (42.6 ± 1.4% and 46.6 ± 0.9%, respectively) than with CSF2 or controls (40.6 ± 1.3% and 39.0 ± 1.3%, respectively). The apoptotic index of the blastocysts was lower (P < 0.05) for DKK1, CSF2 and DKK1+CSF2 groups (3.44 ± 0.14, 3.39 ± 0.11 and 3.11 ± 0.22, respectively) compared to controls (6.64 ± 0.25), and was similar to that of the IVF blastocysts (3.67 ± 0.18). Although the total cell number was similar for the DKK1, CSF2, DKK1+CSF2 and control groups (200.4 ± 3.05, 196.4 ± 3.73, 204.7 ± 3.71 and 205 ± 4.03, respectively), the inner cell mass:trophectoderm cell number ratio of DKK1, CSF2 and DKK1+CSF2 groups (0.21 ± 0.01, 0.17 ± 0.01 and 0.22 ± 0.02, respectively) was higher (P < 0.05) than controls (0.13 ± 0.01) and was similar to that of IVF blastocysts (0.19 ± 0.01). Treatment with DKK1 or CSF2 or both increased (P < 0.05) the expression level of OCT4, NANOG,SOX2, GATA6, BCL2, PTEN, P53, FGF4, GLUT1 and IFN-τ, and decreased that of C-MYC, CDX2, CASPASE, DNMT3a, TCF7 and LEF1 in blastocysts, compared to controls. Transfer of DKK1-treated embryos to 13 recipients resulted in 4 pregnancies (30.8%; 2 live births, one abortion and one currently at 9 months of pregnancy) whereas, transfer of DKK1+CSF2-treated embryos to 16 recipients, resulted in 4 pregnancies (25.0%), all of which resulted in live births. No pregnancy was obtained after transfer of control and CSF-treated embryos to 12 and 16 recipients, respectively. These results suggest that DKK1 treatment of NT embryos increases the blastocyst, conception and live birth rate, and improves their quality whereas, CSF2 treatment, does not affect the blastocyst, conception and live birth rate despite improvement in embryo quality.

Testing the endometrium: is there enough evidence to justify clinical use?

Embryo implantation remains the limiting factor in assisted reproduction outcomes. To date research has mainly focused on improving embryo quality, numbers and selection as the route to improve treatment results. However, with success rates plateauing, interest in the possibility of modulating the endometrial factor is increasing, and a number of biomarkers are now available that offer the possibility of assessing endometrial function. In this review, we review recent evidence for the efficacy of a number of these biomarkers, with emphasis on those that aim to enable improvement in embryo/endometrial developmental synchrony endometrium and that offer an assessment of the degree of immune activation of the endometrium. The emerging field of reproductive tract microbiome analysis is also considered. Finally, nascent biomarkers of materno-foetal dialogue, including noncoding RNAs, microvesicles and endometrial glycans are discussed. Tests of potential clinical value are emerging, but further validation studies are required. The usage of innovative endometrial biomarkers provides the possibility of targeted therapies rather than the blind empirical approaches to face embryo implantation failure. It also enables the possibility of randomized controlled trials of interventions targeting the individual cause rather empirical treatments of undiagnosed recurrent implantation failure.

L-carnitine supplementation in culture media improves the pregnancy rate of in vitro produced embryos with sexed semen from Bos taurus indicus cows.

The in vitro embryo production industry in the actual world presents some difficulties related to low embryonic production rates, a problem that could be associated with in vitro culture conditions that differed from the in vivo (oviductal) conditions, mainly related to cytoplasmic lipid accumulation. L-carnitine is known as a modulator of β-oxidation in the developing embryo, as it has been demonstrated that it improves embryo quality without affecting the in vitro embryo production rate. The aim of the present work was to evaluate the effect of L-carnitine supplemented during the in vitro maturation and culture processes on the implantation rate of in vitro produced embryos. Supplementation with 3.8mM of L-carnitine was used during in vitro maturation, and later, during late in vitro culture, it was added at 1.5mM. A control group contained no L-carnitine supplementation. Bovine oocytes obtained by ultrasound-guided follicle aspiration from healthy Bos taurus indicus cows were matured, fertilized and cultured in vitro. Multiparous F1 (Bos taurus taurus × Bos taurus indicus) cows were used as recipients. Overall, 460 oocytes were processed in three independent replicates from in vitro maturation until day 8 of the in vitro culture. No significant difference was found between treatments of in vitro embryo production. However, pregnancy rate at days 45 and 72 was significantly higher in blastocysts derived from L-carnitine treatment (31.55 ± 9.78%) compared to the control group (18.68 ± 6.31%). In conclusion, addition of L-carnitine at 3.8mM and 1.5mM in the maturation, and culture medium after day 3 of in vitro production process, significantly improved pregnancy rate after embryo transfer.

Myo-Inositol's Role in Assisted Reproductive Technology: Evidence for Improving the Quality of Oocytes and Embryos in Patients With Polycystic Ovary Syndrome.

Polycystic ovary syndrome (PCOS) is one of the most common causes of subfertility, and it is characterized by hormonal dysregulation like insulin resistance. Various measures have been taken in the past to overcome this insulin resistance to improve fertility treatment outcomes. The current paper aims to review and compare the existing studies and literature to assess the impact of myo-inositol (MI) on oocyte and embryo quality in assisted reproductive technology (ARTs). We thoroughly searched the PubMed and Google Scholar databases by using the keywords “PCOS, polycystic ovarian syndrome, inositol, oocyte quality, embryo quality, assisted conception, ART, IVF, and in vitro fertilization.” Nine articles were finalized for review in this paper. Many of the reviewed studies have shown a trend toward the improvement of embryo quality in women with PCOS after MI supplementation; however, there is a lack of statistically significant evidence to support the use of MI in enhancing the quality of oocyte and/or embryo. Clear evidence regarding the role of MI in enhancing the quality of oocyte and embryo in PCOS is limited. A well-controlled, large, randomized controlled trial is required to definitively accept or refute its role.

New Approach of Hyaluronic Acid Bound Spermatozoa-ICSI in Iraqi Infertile Patients

Background: Ability of spermatozoa to binding to hyaluronan is related to sperm membrane maturity and fertilizing potential, thus it has been suggested that sperm selection using Hyaluronic acid (HA) could increase the implantation rate in Intracytoplasmic Sperm Injection(ICSI). Basically Physiological -ICSI (PICSI) is a scientific technique used in ICSI, is a specialized form and more filtered way of selecting the best competent sperm for fertilization process. Despite the fact that conventionally spermatozoa are selected for ICSI based on their morphology and motility. Objective: To evaluate whether (PICSI) has the potential to improve the fertilization rate and embryo grading. Patients, Materials and Methods: Fifty-three infertile couples subjected to an in vitro fertilization stimulation program; Thirty-five underwent (ICSI) cycle and eighteen underwent (PICSI) cycle, assessed the fertilization rate and embryo grading at cleavage stage then the results were statistically analyzed. Results: In spite of the fertilization rate of PICSI group was better than ICSI group but statistically no significant difference was noticed between both of them. In regarding to embryo grading at cleavage stage, there was a significant increase(P&lt;0.05) in Grade 1 and Grade 2 for PICSI group as compared to ICSI group. Conclusions: The percentage of fertilization for PICSI group is better than for ICSI group. In regarding to statistical analysis, there was no significant differences between two groups. Regarding to embryo quality at cleavage stage, the PICSI cycle significantly improves embryo quality(Grade 1 and Grade 2) at day 2 and day 3 of development as compared to the ICSI cycle.

IMD/ADM21-47, a factor that improves embryo quality

Starting in vitro fertilization process with competent oocytes that may endure first cellular divisions is a critical step for obtaining an embryo. To obtain in vitro competent oocytes, culture conditions should emulate the in vivo microenvironment as close as possible. With the aim of improving the in vitro culture medium, the present study evaluated the IMD/ADM21-47 peptide as a factor that promotes oocyte competence and improves embryo quality in bovine systems. The culture supplemented with 153 μg/mL of IMD/ADM21-47 was correlated with the production of healthy oocytes in metaphase II (MII) stage in compacted cumulus-oocyte complexes (COC) with a decrease of BAX/BCL-2 to mRNA ratio and a reduction of late apoptosis by TUNEL in MII oocytes. In addition to this, treatment with IMD/ADM21-47 caused cAMPi level to be constant over time, and the cAMPi level kept increasing until 6 h. COC supplementation with 153 μg/mL of IMD/ADM21-47 increased the blastocyst production rate two-fold in comparison with control conditions. Only embryos from COC treatment with this peptide were capable of developing blastocysts in stage-6 grade I; compared with the control culture, it was the treatment with the greater number of blastocysts stage-5; these are characteristics of good quality blastocysts.

Influence of allicin administration on reproductive efficiency, immunity and lipid peroxidation of rabbit does under high ambient temperature.

This study investigated the effect of daily oral administration with allicin levels (0, 5 and 10mg/kg of female body weight), 30days pre-insemination, on reproductive performance in vivo and in vitro, immunity, and oxidative stress of rabbit does under high ambient temperature. Niliparous NZW does (n=105) were randomly divided into three groups (35 in each) treated with 0, 5 and 10mg allicin dissolved in 2ml distilled water, respectively, for 30days pre-insemination. At the end of treatment (30days), does were artificially inseminated with fresh diluted semen of 20 fertile NZW bucks. Reproductive performance and ovulatory response parameters were determined. Serum biochemicals, enzyme activity, immunoglobulins (IgG and IgM) and antioxidant status were determined on day 30 of treatment. Serum progesterone and prolactin were determined pre-insemination (30days of treatment), on 15days of pregnancy and 7days post-partum. Results showed that both allicin levels increased live litter size, and bunny viability rat and litter size at birth and weaning. Allicin levels increased ovulation rate and improved embryo quality. Number of total follicles decreased only with 10mg allicin. Progesterone increased pre-insemination, 15days of pregnancy and 7days post-partum progesterone by allicin levels. Prolactin pre-insemination and on day 7 post-partum increased with 10mg allicin. Serum total proteins, albumin, globulin, IgG and IgM increased, while glucose, aspartate and alanine aminotransaminases, and thiobarbituric acid reaction decreased by both allicin levels. In conclusion, the mechanism by which allicin administration 30days pre-insemination to improve the reproductive performance of rabbit does is based on that allicin can play an important role, as a natural exogenous antioxidant, increasing immune response and reducing lipid peroxidation.

L-carnitine reduces the adverse effects of ROS and up-regulates the expression of implantation related genes in in vitro developed mouse embryos

In vitro developed embryos are inevitably exposed to various reactive oxygen species (ROS) which may decrease the embryo’s competence in assisted reproductive technology (ART) procedures. Optimization of embryo culture media using antioxidant agents could help to improve embryo quality and could overcome failures in current ART. The aim of this study was to evaluate the effects of l-carnitine (LC), an enhancer of mitochondrial activity and free radical scavenger, in culture media on early embryo competence and expression of ErbB1 and ErbB4 implantation related genes. Two-cell mouse embryos were cultured in the following four conditions: 1. LC group in media containing LC; 2.H 2O2 group exposed to H2O2 for 30 min and then transferred into a simple media; 3.H2O2+LC group exposed to H2O2 for 30 min and then transferred into a simple media containing LC; 4.the control group kept throughout in simple media. All groups were allowed to develop until the blastocyst stage. ErbB1 and ErbB4 expression were evaluated by Real-time PCR and immunocytochemistry. The expression of Sirt3 gene was also evaluated. Intracellular ROS levels were examined by DCFH-DA fluorescence intensity. In order to assess the morphological quality of the embryos, ICM and OCM number blastocyst cells were evaluated by using Hoechst and propidium iodide (PI) staining. ErbB1, ErbB4, ROS levels and cell number were compared across all in vitro groups. Our data reveal that LC significantly increases ErbB1 and ErbB4 gene and protein expression with intracellular ROS levels and Sirt3 gene expression significantly decreased after LC treatment. It is worth noting that an elevated cell number was observed in the LC-treated group compared with the other groups suggesting increased viability and/or proliferation. Our findings suggest that the use of LC could be helpful to improve preimplantation embryo culture media through its effects in decreasing ROS levels and the increase of implantation-related genes.

The Effect of Recombinant LH Addition to Recombinant FSH on Assisted Reproductive Technologies Outcomes in Overweight and Obese Patients without Polycystic Ovary Syndrome

Background: Obesity and overweight negatively affect fertility. Protocols need to be developed in ART treatments for obese and overweight women. Objectives: To investigate the effect of recombinant luteinizing hormone (rLH) addition to recombinant follicle stimulating hormone (rFSH) on treatment outcomes in assisted reproductive technologies (ART) cycles in obese and overweight women without polycystic ovary syndrome. Methods: This retrospective cohort study was carried out in Kocaeli University Faculty of Medicine, Assisted Reproductive Techniques Clinic between January 2016 and March 2019. To analyze the impact of rLH addition to rFSH in GnRH antagonist cycles in overweight and obese, the patients were divided into four groups according to body mass index (BMI) and gonadotropin type; Group 1: patients with BMI ≥ 25 stimulated with rFSH alone (n: 37), Group 2: patients with BMI ≥ 25 stimulated with rFSH + rLH (n: 37), Group 3: patients with BMI between 18.5 - 24.99 stimulated with rFSH alone (n: 33), Group 4: patients with BMI between 18.5 and 24.99 stimulated with rFSH + rLH (n: 30). Patients with polycystic ovary syndrome were excluded. Results: Basal LH levels were found to be significantly lower in obese and overweight patients compared to normoweight patients (p = 0.01). Grade 1 embryo ratio in obese and overweight patients was higher in rLH added obese group than in group LH not included (64.9%, p = 0.005). Ongoing pregnancy rates (OPR) in obese and overweight patients were significantly higher in rLH added group compared to rFSH only group (43.2% vs 18.9% respectively, p = 0.044). However, OPR did not differ significantly in rLH added and rFSH only groups in normoweight patients (p = 0.588). Conclusion: This study has shown that obese and overweight non-PCOS patients have lower endogenous LH levels. It has also shown that rLH supplementation in GnRH antagonist cycles in obese and overweight women improves embryo quality and ongoing pregnancy rates. However rLH addition to rFSH doesn’t seem to have a value in normoweight patients.

Effect of a 6-week “Mediterranean” dietary intervention on in vitro human embryo development: the Preconception Dietary Supplements in Assisted Reproduction double-blinded randomized controlled trial

To study the impact of increased dietary intake of omega-3 fatty acids, vitamin D, and olive oil for 6 weeks before invitro fertilization (IVF) or IVF-intracytoplasmic sperm injection (ICSI) on morphokinetic markers of early embryo development. A double-blinded randomized controlled trial. Academic IVF unit. A total of 111 couples undergoing IVF or IVF-ICSI were recruited. Fifty-five couples received the 6-week study intervention of a daily supplement drink enriched with omega-3 fatty acids and vitamin D plus additional olive oil and olive oil-based spread, and 56 couples received the control intervention. The primary end point for the study was the time taken for completion of the second cell cycle after fertilization (CC2). Secondary end points included time to complete the third and fourth cell cycles (CC3 and CC4), the synchrony of the second and third cell cycles (S2 and S3), and the day 3 and day 5 Known Implantation Data Scores (KIDScores). There was no difference in CC2 between the two groups. However, CC4 was accelerated in the study group compared with the control group, and a significantly shortened S3 as well as an increase in KIDScore on day 3 were observed, indicating improved embryo quality in the study group. This study demonstrates that a short period of dietary supplementation alters the rate of embryo cleavage. Further research is required to investigate the mechanisms that regulate this effect, and whether the impact on embryo development translates into improved clinical outcomes. ISRCTN50956936.