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.

Prediction of the in vitro developmental competence of early-cleavage-stage human embryos with time-lapse imaging and oxygen consumption rate measurement.

PURPOSE: To assess an embryo's ability to develop into a good-quality blastocyst during the early-cleavage stage using time-lapse imaging and the oxygen consumption rate. METHODS: In total, 942 zygotes had their oxygen consumption rates measured. In total, 282 zygotes were assessed by using time-lapse imaging. In total, 121 zygotes were examined by using both their oxygen consumption rate and time-lapse imaging. RESULTS: The embryos with moderate respiration rates of between 0.41 and 0.61 (×1014/mol s-1) on day 3 had a 22.1% chance of becoming good-quality blastocysts; those outside that range had a 14.3% chance. With the time-lapse system, when the first division was within 24 hours, 22.3% of the embryos grew to good blastocysts. After 24 hours, the rate dropped to 8.6%. The intervals between two consecutive cleavages were calculated and the duration of the second cell cycle was defined. When the time was between nine hours and 13 hours, there was a higher rate of good blastocysts. Regarding both criteria, when the embryos had progressed in the optimal range, a high percentage of them had become good blastocysts; it was 8.0% outside of that range. CONCLUSION: Individual embryos with the potential to develop into good-quality blastocysts could be selected at day 3 of culture using these systems.

Effects of cyclophosphamide administration on the in vitro fertilization of mice.

PURPOSE: To evaluate the oocyte fertilization ability and embryo growth after cyclophosphamide (CPA) treatment in mice. METHODS: Mice were treated with CPA at different doses (0-800 mg/kg body weight). The oocytes then were retrieved and evaluated for their in vitro fertilization efficiency. RESULTS: The average number of metaphase II (MII) oocytes significantly decreased by ≥400 mg/kg CPA administration. The fertilization rate also decreased in the group that was treated with ≥400 mg/kg CPA. However, after fertilization, the embryos demonstrated normal growth ability. Two weeks after CPA administration, the number of mice from which the oocytes could be retrieved markedly decreased, but the fertilization rate and development of morphological features in the embryos were similar to those of the controls. One month after CPA administration, the number of mice from which the oocytes could be retrieved, fertilization rate, and development of the morphological features in the embryos were similar to those of the controls. CONCLUSION: The number of oocytes decreased as the CPA administration level increased; however, the oocytes' potential for fertilization and development to the blastocyst stage was not significantly affected. One month after CPA administration, the number of oocytes and the potential for development into blastocysts were recovered.

The efficacy of the transfer of twice frozen-thawed embryos with the vitrification method.

OBJECTIVE: To confirm the clinical benefits of recryopreserved, twice-thawed embryo transfer (ET). DESIGN: Retrospective study. SETTING: Private fertility clinic. PATIENT(S): Forty-nine women whose embryos had been refrozen after a previous frozen-thawed ET. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Comparison of implantation and pregnancy rates of twice-cryopreserved, twice-thawed embryos versus once-cryopreserved, once-thawed embryos. RESULT(S): The pregnancy rate per ET cycle was 27.8% in the refrozen group and 25.9% in the control group (no statistically significant difference). The implantation rate was 25.0% in the refrozen group and 19.3% in the control group (no statistically significant difference). CONCLUSION(S): The refreezing of supernumerary embryos can prevent ovarian hyperstimulation syndrome in stimulated patients and in those who have experienced repeated failed pregnancies. If unexpected supernumerary embryos are available for recryopreservation after frozen-thawed ET, these embryos may be revitrified for a future transfer.

Regulation of amphiregulin, EGFR-like factor expression by hCG in cultured human granulosa cells.

BACKGROUND: It has long been suspected that the epidermal growth factor (EGF) receptor and some of its putative ligands may play an important role in ovarian function. Amphiregulin (AR) is the growth factor with an EGF-like motif, but its potential role in signalling in the ovary is still obscure. AR gene expression and its functional effect were evaluated in human granulosa cells from immature follicles. METHOD: Granulosa cells from immature follicles with early menstrual phase were cultured with or without 200 mIU/ml of FSH stimulation, following with or without 1 IU/ml of hCG. mRNA levels of AR and luteinising hormone replacement (LHR) were semi-quantified using RT-PCR. Progesterone (P) concentration in the medium was assayed. RESULTS: LHR mRNA was expressed 48 h after FSH stimulation without AR mRNA expression. AR mRNA was expressed 1 h after hCG stimulation, and increased the intensity in 6 h. P biosynthesis was increased by AR in a dose-dependent manner. AR mRNA was elevated by forskolin stimulation without FSH and hCG stimulation before LHR mRNA expression. When cultured with FSH for 15 h, followed by increasing doses of hCG stimulation for 6 h, the AR mRNA levels increased according to hCG concentration up to 1,000 mIU/ml. CONCLUSION: Occurrence of LHR gene expression following FSH stimulation was necessary for the AR gene expression in vivo, and the AR gene was induced by forskolin without LHR gene expression in vitro. P biosynthesis was stimulated, to some extent, by AR. This result suggests the differentiation effect of AR on granulosa cells. AR might be a mediator of LH signals before ovulation.

Successful pregnancy after the vitrification of zygotes using commercial vitrification solutions and conventional straws to protect against infections in liquid nitrogen.

PURPOSE: To report on successful birth after the transfer of postthawed human zygotes that were vitrified using a conventional straw for the purpose of protecting them from infections and a low-toxicity cryoprotectant that is commercially sold. METHODS: A primary infertile couple presented at our IVF program. After being checked for fertilization, the embryos were not transferred to the uterus at that cycle. Instead, all of them were cryopreserved at the 2-pronuclei stage using our original vitrification method. After the vitrification and warming of four zygotes, two embryos were transferred into the uterus. RESULTS: Twenty-one 2-pronuclei embryos were vitrified in liquid nitrogen. After 2 embryos were thawed and transferred, successful pregnancy was the outcome, and a healthy boy was born at term. CONCLUSIONS: Vitrification is a simple procedure and requires less time than slow freezing. Vitrification of zygotes in a conventional straw seems to be sufficient for viability and works to store the zygotes safely.

Comparison of sibling oocyte outcomes after intracytoplasmic sperm injection and in vitro fertilization in severe teratozoospermic patients in the first cycle.

To follow up the outcome of sibling oocytes subjected to both conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) in the first cycles of severe teratozoospermic patients with normal sperm morphology (NSM)or=6 cell embryos on day 3 and blastocyst formation on days 5 and 6, did not differ significantly between the two groups. There was a trend towards a high pregnancy rate cycle in mixed ICSI/IVF embryo transfer (ET) (49.1%). In conclusion, this study showed that in couples with only severe teratozoospermia, there was a benefit in subjecting sibling oocytes to both IVF and ICSI in the first cycle because 24 (28.2%) cycles of total fertilization failure were avoided. Furthermore, despite initially significant higher fertilization rates in ICSI than IVF oocytes, subsequent rates of development from >or=6 cells up to blastocyst stage were the same suggesting that ICSI should be used with caution, as after day 3, ICSI-derived embryo development was compromised compared with IVF.

Embryo outcome in Y-chromosome microdeleted infertile males after ICSI.

UNLABELLED: A prospective study involving 118 infertile Japanese couples to assess the embryo outcomes in both azoospermic and oligoasthenoteratoazoospermic (OAT) patients with Y-chromosome microdeletion. The men were divided into two groups; azoospermia (n = 27), and OAT, sperm concentration <5 x 10(6)/ml (n = 91). They were investigated for Y-chromosome microdeletions by a polymerase chain reaction (PCR) amplification of the Y-chromosome-specific sequence tag site (STS). The embryo outcomes of patients found to have Y-microdeletion were determined. The frequency of microdeletion was 8.8% (9) and two had microdeletions distal to DAZ. The mean fertilization rate and the cleavage rate in the eight cycles of both azoospermic and oligospermic patients were 59.3 and 87.5%, respectively. The percentages of grade 1 & 2 embryos, > or =6 cells embryos, and blastocyts were 51.7, 65.6, and 45.3%, respectively. Three pregnancies resulted from the eight cycles (37.5%). CONCLUSION: in Y-chromosome microdeletion cycles in which sperm cells were available for intracytoplasmic sperm injection (ICSI), embryo outcome was comparable to conventional IVF.