Creation, effects on embryo quality, and clinical outcomes of a new embryo culture medium with 31 optimized components derived from human oviduct fluid: A prospective multicenter randomized trial.

Purpose: Our aim is to make an ideal embryo culture medium close to human oviduct fluid (HOF) components, and to evaluate the quality of this medium with embryo quality and clinical outcomes in assisted reproductive technology (ART) by a prospective randomized controlled trial (RCT). Methods: Study I: HOF was collected laparoscopically from patients (n = 28) with normal pelvic findings. According to HOF analysis results, the new medium "HiGROW OVIT®" (OVIT) was designed. Study II: Embryos (2 pronuclei (2PN) = 9633) were assigned from 1435 patients. The blastulation rate (BR), good BR (gBR), utilized (transferred/cryo-preserved) BR (uBR), pregnancy rate (PR), and miscarriage rate (MR) were compared between the OVIT and control groups by RCT. Results: The novel medium 'OVIT' was produced according to 31 HOF components. The concentrations of essential amino acids (e-AAs) were lower in OVIT than in current media, yet the opposite was true for ne-AA concentrations. gBR and uBR were higher in the OVIT group than in the control group. In the older female group, gBT and uBR were significantly higher in the OVIT group. Conclusions: The novel medium 'OVIT' was produced according to HOF data. The OVIT had significantly better embryo quality and clinical outcomes than the current media.

In vitro fertilization embryo development from caffeine-treated murine sperm.

Purpose: To evaluate the effect of long-term caffeine administration on murine sperm and subsequent in vitro fertilization (IVF). Methods: Male mice were injected with various doses (0, 0.2 and 1.0 mg/mouse/day) of caffeine for 1 month. After sperm collection, the IVF rate and embryo development to the blastocyst stage were evaluated. Results: The mean body weight significantly decreased in the 1.0 mg/day treatment group compared to the control group (P < 0.01). Testicular weight and histological features did not differ, and total blood testosterone was no different in spite of the difference between 0.2 and 1.0 mg/day of caffeine. The IVF rate differed significantly between the control group [100/105 (95.2 %)] and 0.2 mg/day group [106/121 (87.6 %)] (P < 0.05). Furthermore, blastocyst formation was significantly and dose-dependently lower with higher caffeine levels: control group: 85/100 (85.0 %); 0.2 mg/day group: 84/106 (79.2 %) (P < 0.05); 1.0 mg/day group: 64/102 (62.7 %) (P < 0.001). Conclusions: Caffeine treatment affected body weight of male mice. However, testicular weight, histological features and total blood testosterone concentration were not statistically different. In addition, following IVF using sperm from these mice, blastocyst formation decreased in a dose-dependent manner. These findings suggest that embryo development from oocytes fertilized with sperm from caffeine-administered male mice is negatively affected.

Effect of long-term caffeine administration to mice on in vitro fertilization and embryo development using oocytes.

PURPOSE: To evaluate the effect of long-term caffeine administration to mice on in vitro fertilization (IVF) of oocytes. METHODS: Mice were injected with different dosages (0, 0.1, and 1.0 mg/mouse/converted day) of caffeine for one month. Subsequently, the fertilization rate and embryo development to blastocyst stage were evaluated in IVF using oocytes from the mice. RESULTS: The retrieved average oocyte rate was significantly lower (27.4) in mice injected with 1.0 mg caffeine than in the control group (36.5; P < 0.05); the fertilization rate was significantly different between the 0 mg (317/401; 79.1 %) and 1.0 mg group (199/301; 66.1 %) (P < 0.05). At 96 h after insemination, the blastocyst formation rate was significantly decreased in the 1.0 mg group (94/199; 47.2 %) compared with the control (0 mg) group (237/317; 74.8 %) and 0.1 mg group (226/323; 70 %) (P < 0.05). When 1.0 mg caffeine was administered for two weeks, embryo development was significantly impacted. CONCLUSIONS: Our findings suggest that caffeine administration negatively impacts oocytogenesis and embryonic development after IVF.

Enhancing the fertility of poor responders by treatment with estrogen rebound combined with a short protocol.

Aim: To assess the efficacy of estrogen rebound (ER) plus flare-up by gonadotropin releasing hormone agonist (GnRH-a) in poor responders who failed to become pregnant prior to a long protocol treatment. Methods: The patients comprised of thirty-one infertile patients with oocyte retrieval levels of less than five, who had undergone several long protocol treatment cycles. The efficacy of treatment with the ER plus flare-up from GnRH-a was compared with the prior long protocol treatment. The main outcome measures are: confirmation of ER, maximal serum E2 levels prior to human chorionic gonadotropin (hCG) administration, follicular development, dose, and duration of gonadotrophins in a clinical setting. Results: The ER was confirmed by estrogen levels; FSH increased with ER plus flare-up from GnRH-a. Although the 31 patients included in the study had undergone frequent prior treatment cycles, including the long protocol, the pregnancy rate per embryo transfer following ER plus flare-up by GnRH-a was 37.5% (nine of 24). The number of follicles, number of oocytes retrieved, and the E2 level was higher than those found in prior treatment cycles. Conclusion: Exogenous estrogen administration with PremarinR plus flare-up by GnRH-a may represent an alternative and effective protocol for poor responder patients who had previously undergone several prior long protocol treatments. (Reprod Med Biol 2003; 2: 127-131).

Relationship between follicular fluid hormone levels, embryo quality, and maternal age during in vitro fertilization after the short or long protocol with a gonadotropin releasing hormone agonist.

Purpose: To examine the relationship between embryo quality and follicular fluid hormonal level in short and long protocol gonadotrophin releasing hormone agonist treatment cycles. Methods: A total of 90 patients had non-polycystic ovary syndrome (non-PCOS) and 10 had PCOS. A total of 100 subjects underwent in vitro fertilization (IVF). Thirty-six subjects underwent conventional IVF and 64 subjects underwent intracytoplasmic sperm injection (ICSI). The dominant follicles were initially retrieved and a hormonal assay was done. A total of 32 patients underwent a short protocol and 66 patients were treated with the long protocol. Estradiol (E2), progesterone (P4), total testosterone (TTE) and androstenedione (ASG) levels in follicular fluid (FF) were compared in the two treatment groups (short and long protocol), in regard to maternal age and oocyte/embryo quality. Results: The retrieval FF volume was not significantly different between the PCOS and non-PCOS patients; however, P4 was significantly lower with PCOS (P < 0.01). Analysis of four different hormone levels was not significantly different between the short and long protocol groups. No significant relationship was found between four hormone levels in regard to oocyte morphology and embryo quality. The levels of P4 of younger women was significantly lower than that of older women; furthermore, a significantly higher TTE and ASG were found in the younger women. Progesterone was found to statistically significantly increase with FF volume. Conclusion: Follicular fluid P4 from the younger group was significantly lower, and TTE and ASG was significantly higher when compared to the older group. Analysis of four different hormone levels revealed no significant difference between the short and long protocol groups. No significant relationship was found between four hormone levels, oocyte morphology, and embryo quality. (Reprod Med Biol 2003; 2: 165-169).