Day-3 vs. Day-5 fresh embryo transfer.

OBJECTIVE: Embryo transfer on day-5 has been associated with higher success rates, therefore our IVF clinics started to extend embryo culture until blastocyst stage. This study aimed to compare the success rates of day-3 vs. day-5 embryo transfers. METHODS: We had 266 patients included, all having undergone ICSI, with 221 patients having undergone day-3 embryo transfers, and 45 patients having undergone day-5 embryo transfers. Patients with more than five good quality embryos on day-3 were chosen to prolong the culture of embryos into day-5. RESULTS: There were no significant differences in patient characteristics, including baseline LH, FSH, Prolactin and Estradiol hormone levels. In addition, there were also no significant differences in rFSH total dosage and duration of stimulation day. Final estradiol levels, number of follicles, retrieved oocytes, matured oocytes, fertilized oocytes and number of embryos were significantly higher in day-5 compared to day-3 embryo transfer groups. Number of embryos transferred on day-3, were significantly higher compared to day-5. Neither group showed any significant differences in clinical pregnancy, implantation, multiple pregnancy or living birth rates. There were no differences in birth weights and lengths, head circumstances and Apgar Scores between both groups either in singleton or twin group. CONCLUSIONS: Transferring embryos at day-3 may provide the same benefits as day-5 embryo transfers to patients. However, more embryos were required to be transferred to achieve these comparable results.

Impact of body mass index on intracytoplasmic sperm injection in women with polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is a condition that affects fertility. There are two types of PCOS; the normal/lean type and overweight/obese type. The aim of this study was to assess baseline characteristics, ovarian response, quality of oocytes, embryos, pregnancy, implantation and live birth rates in normal/lean and overweight/obese patients with PCOS undergoing ICSI compared with patients without PCOS. This retrospective case-control analytical study included 38 normal/lean and 17 overweight/obese patients with PCOS, and 98 normal/lean and 17 overweight/obese patients without PCOS. Parameters were observed based on baseline characteristics, ovarian response to dosage and duration of gonadotropin administered, number of oocytes, matured oocytes, fertilization rate, embryo quality and development, pregnancy, implantation and live birth rates. Basal serum luteinizing hormone in normal/lean PCOS was significantly higher compared with non-PCOS groups. Total dosage of gonadotropin used was significantly lower in normal/lean PCOS compared with other groups. End estradiol levels in normal/lean PCOS was significantly higher compared with the non-PCOS groups. Number of follicles, retrieved oocytes and matured oocytes were significantly higher in PCOS groups compared with the non-PCOS groups. However, there were no differences in fertilized oocytes, cleavage, number of top-quality embryos, pregnancy, implantation, and live birth rates among groups. This present study suggests that normal/lean PCOS requires lower gonadotropin dosages and that patients with PCOS have more follicles and oocytes compared with patients without PCOS, however the number of fertilized oocytes and embryos from patients with PCOS were the same as those from patients without PCOS and suggested that the quality of retrieved oocytes in PCOS might be compromised.

Deleterious effect of short-term gavage of an ethanol extract of cogon grass (Imperata cylindrica L.) roots on testis and epididymal sperm quality.

BACKGROUND AND AIM: Cogon grass (Imperata cylindrica L.) (CGG) is a herbal medicine that could be developed into a male antifertility agent. The present study aims to determine the effect of an ethanol extract of CGG roots on mice testicular activity, reproductive hormone levels, and epididymal sperm quality. MATERIALS AND METHODS: This study was designed as completely randomized with three different doses, such as an ethanol extract of CGG roots at 0 (control), 90, and 115 mg/kg body weight. In total, 21 male DDY mice strain were treated with the CGG extract (by gavage) for 14 days, followed by an evaluation of reproductive organs, epididymal sperm quality, testis histology, histomorphometry, and reproductive hormone assays. All quantitative data were analyzed by analysis of variance, followed by Tukey's post hoc test at α=0.05. RESULTS: The results showed that the administration of the CGG root ethanol extract disrupted the testis interstitial area and seminiferous tubules, resulting in decreased epididymal sperm quality as well as serum testosterone levels in a dose-dependent pattern. CONCLUSION: Oral administration of a CGG root ethanol extract induced testicular damage, decreased epididymal sperm quality, and impaired testosterone secretion.

Tolerance of lamb and mouse oocytes to cryoprotectants during vitrification.

SummaryMouse and lamb oocytes were vitrified with, or exposed to, different cryoprotectants and evaluated for their effects on their survival and developmental competence after in vitro fertilization (IVF) and activation treatments. Control oocytes remained untreated, whilst the remainder were exposed to three different combinations of vitrification solutions [dimethyl sulfoxide (DMSO) + ethylene glycol (EG), EG only, or propanediol (PROH) + EG] and either vitrified or left unfrozen (exposed groups). Oocytes in the control and vitrified groups underwent IVF and developmental competence was assessed to the blastocyst stage. In lambs, survival rate in vitrified oocytes was significantly lower than for oocytes in the exposed groups (P <0.05). Blastocyst development was low in vitrified oocytes compared with controls (<6% vs 38.9%, P <0.01). Parthenogenetic activation was more prevalent in vitrified lamb oocytes compared with controls (P <0.05). No evidence of zona pellucida hardening or cortical granule exocytosis could account for reduced fertilization rates in vitrified lamb oocytes. Mouse oocytes demonstrated a completely different response to lamb oocytes, with survival and parthenogenetic activation rates unaffected by the vitrification process. Treatment of mouse oocytes with DMSO + EG yielded significantly higher survival and cleavage rates than treatment with PROH + EG (87.8% and 51.7% vs 32.7% and 16.7% respectively, P <0.01), however cleavage rate for vitrified oocytes remained lower than for the controls (51.7% vs 91.7%, P <0.01) as did mean blastocyst cell number (33 ± 3.1 vs 42 ± 1.5, P <0.05). From this study, it is clear that lamb and mouse show different tolerances to cryoprotectants commonly used in vitrification procedures, and careful selection and testing of species-compatible cryoprotectants is required when vitrifying oocytes to optimize survival and embryo development.

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.

Bone morphogenetic protein 15 in the pro-mature complex form enhances bovine oocyte developmental competence.

Developmental competence of in vitro matured (IVM) oocytes needs to be improved and this can potentially be achieved by adding recombinant bone morphogenetic protein 15 (BMP15) or growth differentiation factor (GDF9) to IVM. The aim of this study was to determine the effect of a purified pro-mature complex form of recombinant human BMP15 versus the commercially available bioactive forms of BMP15 and GDF9 (both isolated mature regions) during IVM on bovine embryo development and metabolic activity. Bovine cumulus oocyte complexes (COCs) were matured in vitro in control medium or treated with 100 ng/ml pro-mature BMP15, mature BMP15 or mature GDF9 +/- FSH. Metabolic measures of glucose uptake and lactate production from COCs and autofluorescence of NAD(P)H, FAD and GSH were measured in oocytes after IVM. Following in vitro fertilisation and embryo culture, day 8 blastocysts were stained for cell numbers. COCs matured in medium +/- FSH containing pro-mature BMP15 displayed significantly improved blastocyst development (57.7±3.9%, 43.5±4.2%) compared to controls (43.3±2.4%, 28.9±3.7%) and to mature GDF9+FSH (36.1±3.0%). The mature form of BMP15 produced intermediate levels of blastocyst development; not significantly different to control or pro-mature BMP15 levels. Pro-mature BMP15 increased intra-oocyte NAD(P)H, and reduced glutathione (GSH) levels were increased by both forms of BMP15 in the absence of FSH. Exogenous BMP15 in its pro-mature form during IVM provides a functional source of oocyte-secreted factors to improve bovine blastocyst development. This form of BMP15 may prove useful for improving cattle and human artificial reproductive technologies.