Differences in clinical outcomes between men with mosaic Klinefelter syndrome and those with non-mosaic Klinefelter syndrome.

Purpose: This study compared the clinical outcomes of men with Klinfelter syndrome based on karyotype. Methods: The authors analyzed the outcomes of microdissection testicular sperm extraction (micro-TESE) performed on 57 patients with Klinfelter syndrome (KS) at our clinic. Results: The average ages of the non-mosaic and mosaic KS groups were 32.2 ± 4.8 and 45.9 ± 13.1 years, respectively. The sperm retrieval rates of the non-mosaic and mosaic KS groups were 46.5% (20/43) and 50.0% (7/14), respectively. The fertilization rates after intracytoplasmic sperm injection did not significantly differ between the non-mosaic and mosaic KS groups. The mosaic KS group had higher cleavage and blastocyst development rates than the non-mosaic KS group (72.2% vs. 96.2% and 30.5% vs. 44.7%, respectively). The group using motile sperm had better outcomes than the group using immotile sperm. The embryo transfer outcomes of the non-mosaic and mosaic KS groups did not significantly differ (clinical pregnancy rate: 28.0% vs. 20.7%, miscarriage rate: 14.3% vs. 33.3%, production rate per transfer: 22.0% vs. 13.8%, and production rate per case: 58.8% vs. 57.1%). Conclusions: Compared with the non-mosaic KS group, the mosaic KS group had significantly better intracytoplasmic sperm injection outcomes because of the higher utilization rate of motile sperm.

Developmental trajectory of monopronucleated zygotes after in vitro fertilization when they include both male and female genomes.

OBJECTIVE: To examine the cause of monopronucleated zygote (1PN) formation that includes both maternal and paternal genomes. DESIGN: Retrospective cohort study. SETTING: Private fertility clinic. PATIENT(S): A total of 44 1PN and 726 2-pronuclear zygotes from 702 patients were observed using 2 different time-lapse observation systems. INTERVENTION(S): Previously recorded time lapse data were reviewed to examine the mechanism of 1PN formation. MAIN OUTCOME MEASURE(S): The distance between the position of the second polar body extrusion and the fertilization cone or epicenter/starting position of the cytoplasmic wave was measured, and the consequent data were analyzed. Cytoplasmic waves were confirmed using vector analysis software. RESULT(S): The cut-off value for the difference in the distance between the position of the second polar body extrusion and the fertilization cone or the epicenter/starting position of the cytoplasmic wave was 17 µm (AUC: 0.987, 95% CI: 0.976-0.999) for the Embryo Scope and 18 µm (AUC: 0.972, 95% CI: 0.955-0.988) for the iBIS time-lapse observation systems. CONCLUSION(S): In this study, it was found with a high degree of accuracy that a monopronucleus is formed when the fusion of the sperm takes place within 18 µm from the point of the second polar body extrusion. The theoretical chance of 1PN occurrence after in vitro fertilization is 2.7% when the sperm is considered to be fused anywhere in the plasma membrane of an oocyte.

The inclusion of blastomeres into the inner cell mass in early-stage human embryos depends on the sequence of cell cleavages during the fourth division.

The fate of the ICM in humans is still unknown, due to the ethical difficulties surrounding experimentation in this field. In this study we have explored the existing time-lapse recording data of embryos in the early stages of development, taking advantage of the large refractile bodies (RBs) within blastomeres as cellular markers. Our study found that the cellular composition of the ICM in humans is largely determined at the time of the fourth division and blastomeres which cleave first to fourth, during the fourth division from 8 cells to 16 cells, have the potential to be incorporated in the ICM.

Noninvasive embryo selection: kinetic analysis of female and male pronuclear development to predict embryo quality and potential to produce live birth.

OBJECTIVE: To evaluate a noninvasive method of examining euploid embryos, focusing on kinetic analyses, from second polar body extrusion to pronuclear membrane breakdown (PNMBD). DESIGN: Retrospective embryo cohort study. SETTING: Private IVF clinic. PATIENT(S): 213 frozen-thawed single blastocyst transfers. INTERVENTION(S): Fertilized oocytes were recorded by means of time-lapse photography, followed by kinetic analysis of female and male pronuclei (PNs). MAIN OUTCOME MEASURE(S): The differences in size between the 2PNs in embryos resulting in live births compared with those of embryos from failed pregnancies were analyzed according to sequential size from early PN stages to PNMBD. RESULT(S): It was found that the difference in areas between male and female PNs immediately before PNMBD is a better predictor of embryo quality if this difference is below a known cutoff value. The size of male PNs 8 hours before the onset of PNMBD should be larger than female PNs (B). The difference in size between male and female PNs 8 hours before PNMBD should be larger than the difference in their size immediately before PNMBD. When normal embryos were defined using the equation (A∪C)∩B, the birth rates for in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) were 68.1% and 50.0%, respectively. For the remaining embryos, defined as abnormal according to the above criteria, birth rates were 9.4% for IVF and 4.2% for ICSI. CONCLUSION(S): We have developed a method for noninvasive embryo evaluation by means of the kinetic analysis of female and male PN growths. This method should enable us to select embryos that have a higher potential for healthy births.

Piezo-assisted ICSI improves fertilization and blastocyst development rates compared with conventional ICSI in women aged more than 35 years.

PURPOSE: Piezo-assisted intracytoplasmic sperm injection (Piezo-ICSI) is reported to be an effective method for inseminating fragile oocytes compared with conventional ICSI (c-ICSI). However, infertile patient groups suitable for Piezo-ICSI have not been elucidated. This study was conducted to determine age groups suitable for Piezo-ICSI using sibling egg controls inseminated by a well-trained embryologist to reduce technical inequalities. METHODS: A total of 947 matured oocytes were inseminated either by Piezo-ICSI or by c-ICSI in sibling oocytes as controls. Fertilization (2 pronuclei, PN), survival, and blastocyst development rates on day (D) 5 and D6 after insemination were compared between the Piezo-ICSI and c-ICSI groups. Further analyses were applied to groups of women >35 or ≤35 years of age. RESULTS: There were no significant differences in fertilization, survival, or blastocyst development rates between the two insemination treatment groups. However, for women >35 years of age, the fertilization (P = .008) and blastocyst development (P = .016) rates with Piezo-ICSI on D5 and D6 were significantly higher than in those subjected to c-ICSI. CONCLUSIONS: Piezo-ICSI was useful for inseminating oocytes from women >35 years of age.

Is human chorionic gonadotropin supplementation beneficial for frozen and thawed embryo transfer in estrogen/progesterone replacement cycles?: A randomized clinical trial.

Aim: Human chorionic gonadotropin (hCG) is used frequently for luteal support in fresh in vitro fertilization cycles as it induces progesterone secretion from the ovaries after oocyte retrieval and modulates the endometrium for implantation in fresh cycles. In contrast, hCG is not usually used for the transfer of cryopreserved-thawed embryos in estrogen/progesterone replacement cycles because ovulation is suppressed. However, several studies have shown that luteinizing hormone and hCG receptors are present in the human endometrium and that hCG can directly induce the decidualization of endometrial stromal cells in vitro. Thus, this study evaluated whether hCG supplementation can be beneficial for cryopreserved-thawed embryo transfer in estrogen/progesterone replacement cycles. Methods: One-hundred-and seventy-three cryopreserved-thawed embryo transfer cycles with estrogen/progesterone replacement were divided randomly into two groups. Transdermal oestradiol was used in combination with vaginal progesterone suppositories for HR. The embryo transfer was performed on day 17 and/or day 20 of the HR therapy cycle in both groups. In Group A, 3000 IU of hCG was administered on days 17, 20, and 23. In Group B, hCG was not used. Results: There was no significant difference in the average age of the patients, the average number of previous assisted reproductive technology cycles, or the average number of embryo transfers between the two groups. The rates of pregnancy and implantation per embryo were 37.2% and 25.3%, respectively, in Group A and 35.6% and 21.7%, respectively, in Group B. The pregnancy and implantation rates were similar in both groups. Conclusion: Supplementation with hCG is not beneficial for cryopreserved-thawed embryo transfer in estrogen/progesterone replacement cycles.

Potential of zygotes to produce live births can be identified by the size of the male and female pronuclei just before their membranes break down.

Aim: To determine whether there are differences in size between the male and female pronuclei immediately before the pronuclear membrane breakdown (PNMBD) and to evaluate whether pronuclear size differences influence normal birth rates. Methods: Time-lapse photography was used to measure the size of each pronucleus, while the outcome of 71 frozen-thawed single blastocyst transfers in patients receiving hormone therapy was analyzed retrospectively. The pronuclear measurements were made 4 hours before the PNMBD, corresponding to 16-20 hours after insemination or intracytoplasmic sperm injection, and immediately before the PNMBD. The differences in the areas between the pronuclei in the zygotes that were associated with the live births were compared with those that were associated with the failed pregnancies. Results: The average difference in the area between the pronuclei 4 hours before and immediately before the PNMBD in the patients with a live birth was significantly smaller than in the patients with a failed birth. In addition, the average area difference in the patients with a successful birth was significantly smaller when the measurements were made immediately before the PNMBD, compared with the measurements 4 hours before the PNMBD. Such differences were not detected among the patients who did not achieve a birth. Conclusion: The birth of healthy babies resulted from zygotes that contained pronuclei of similar size when the measurements were made immediately before the PNMBD. Evaluating the size of each pronucleus immediately before the PNMBD provides an effective indicator of the embryo's potential at an early stage of development.

Retrospective comparative study of the factors affecting birthweights in frozen-thawed embryo transfer, compared to fresh embryo transfer.

Aim: Several studies have indicated that the cause of the increased birthweight of frozen-thawed embryos was associated with assisted reproductive technology (ART) procedures, such as cryopreservation. In the present study, the mean birthweight of singletons was compared between the ovulatory and hormone replacement therapy (HRT) cycles in order to investigate the primary factor that leads to higher birthweights from frozen-thawed embryo transfer (FET). Methods: This retrospective study was carried out from January 2011 to December 2014 on 2738 singletons who were born at 37-41 weeks' gestation, following ART in a single facility. The mean birthweight of the singletons who were born after a fresh embryo transfer (ET) was compared to the mean birthweight of the singletons who were born after a FET. In the FET cycles, the mean birthweight of the singletons was compared between the ovulatory and HRT cycles. Results: The mean birthweight of the singletons who were born after a FET was significantly higher than that of the singletons who were born after a fresh ET. In the FET cycles, the birthweight from the HRT cycles was significantly higher than that from the ovulatory cycles. Among the HRT cycles, there was no difference between the birthweight of the singleton who were born from a blastocyst transfer and those who were born from a cleavage-stage ET. Conclusion: The primary factor to affect the birthweight of singletons might be the pre/postET hormonal environment of the endometrium and not the stage of the transferred embryo nor the frozen-thawed procedure itself.

Grade and looseness of the inner cell mass may lead to the development of monochorionic diamniotic twins.

OBJECTIVE: To examine the relationship between the inner cell mass (ICM) grade and its morphological configuration on the occurrence of monochorionic diamniotic (M-D) twinning. DESIGN: Retrospective embryo cohort study. SETTING: Private IVF clinic. PATIENT(S): Evaluation of frozen-thawed single blastocyst transfers with hormone replacement treatment in 8,435. This cohort included 71 blastocysts and their ICMs observed by time-lapse photography. INTERVENTION(S): Any changes in configuration of the ICMs observed by time-lapse photography were analyzed retrospectively. MAIN OUTCOME MEASURE(S): The amount of loosening of blastomeres within the ICM was evaluated by time-lapse observations. The number of cells that were involved in the loosening process was also assessed. Both of these parameters were correlated with the type of monozygotic twinning that eventuated. RESULT(S): The M-D twinning incidence resulting from blastocysts with a high grade ICM (grade A) were transferred was 0.38% (3/796), whereas it was significantly higher, 1.38% (34/2,463), when blastocysts with a poorer (B and C) grade ICM were transferred. Among 71 transferred frozen-thawed blastocysts that were studied with time-lapse photography, there were two dichorionic diamniotic and one M-D twins. Careful observations of the embryo that resulted in the one M-D case, revealed that the ICM acquired a looser appearance due to decompaction of at least eight cells. This type of decompaction was not observed in the ICMs of other transferred blastocysts. CONCLUSION(S): The occurrence of M-D twinning may be avoided by excluding blastocysts that contain decompacting ICMs.