Search

Embryo selection is needed to optimize the chances of pregnancy in assisted reproduction technology. This study aimed to validate non-invasive preimplantation genetic testing for aneuploidy (niPGT-A) using a routine IVF laboratory workflow. Can niPGT-A combined with time-lapse morphokinetics provide a better embryo-selection strategy? A total of 118 spent culture mediums (SCMs) from 32 couples were collected. A total of 40 SCMs and 40 corresponding trophectoderm (TE) biopsy samples ( n = 29) or arrested embryos ( n = 11) were assessed for concordance. All embryos were cultured to the blastocyst stage (day 5 or 6) in a single-embryo culture time-lapse incubator. The modified multiple annealing and looping-based amplification cycle (MALBAC) single-cell whole genome amplification method was used to amplify cell-free DNA (cfDNA) from the SCM, which was then sequenced on the Illumina MiSeq system. The majority of insemination methods were conventional IVF. Low cfDNA concentrations were noted in this study. The amplification niPGT-A and conventional PGT-A was 67.7%. Based on this study, performing niPGT-A without altering the daily laboratory procedures cannot provide a precise diagnosis. However, niPGT-A can be applied in clinical IVF, enabling the addition of blastocysts with a better prediction of euploidy for transfer.

Objective: Progesterone elevation on the day of human chorionic gonadotropin (hCG) administration remains one of the most controversial topics in reproductive endocrinology. Factors associated with these increases have not been fully determined. The purpose of our study is to investigate factors associated with extreme progesterone elevation on the day of human chorionic gonadotropin (hCG) administration., Materials and Methods: This retrospective observational, single-center cohort study recruited 2000 fresh in vitro fertilization (IVF) and/or intracytoplasmic sperm injection(ICSI)-embryo transfer cycles from January 2000 to December 2014 in our institution., Results: When cycles were divided into those with progesterone <1.94 ng/mL (n = 1791) and ≧1.94 ng/mL (n = 209) on the day of hCG administration, five factors were positively associated with highly elevated progesterone concentration: protocol (GnRH agonist versus antagonist; odds ratio [OR = 2.786]), number of dominant follicles (OR = 1.098), total dose of follicle stimulating hormone (FSH) used (OR = 1.023), elevated luteinizing hormone (LH) (OR = 1.085) and estradiol (E2; OR = 1.001) concentrations on the day of hCG administration (p < 0.001 each). After omitting the protocol effect, the remaining factors showed limited contributions to highly elevated progesterone (ORs = 0.95-1.2)., Conclusions: The factor showing the greatest association with extreme progesterone elevation was use of the GnRH agonist protocol., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

This study aimed to describe outcomes in four women aged 28-34 years with central cytoplasmic granulation (CCG) of the oocytes who underwent in vitro fertilization/intracytoplasmic sperm injection (ICSI) using gonadotrophin-releasing hormone (GnRH) agonist to replace human chorionic gonadotrophin (hCG) as a trigger of final oocyte maturation. The initial ICSI procedure showed that all four women had CCG of the ooplasm and poor quality embryos. Subsequent ICSI used an antagonist protocol with a GnRH agonist trigger replacing the agonist protocol, plus hCG triggered ovulation. Ooplasm and embryo quality were improved in all four patients. All four became pregnant and gave birth to live infants. This study provides GnRH agonist triggering that may improve ooplasm granularity and embryo quality.

Purpose: To evaluate the relationship between elective blastocyst transfer, freezing strategy, and the abandonment of frozen embryos with a storage time limit of 10 years as specified in the National Assisted Reproduction Act of Taiwan., Methods: This two-phase retrospective cohort study was conducted at a single tertiary center, Kaohsiung Chang Gung Memorial Hospital (KCGMH), in 2019. Participants were selected from a data registry containing 4167 fresh IVF cycles, including phase 1 cycles from 1999 to 2009 and phase 2 cycles from 2010 to 2014, at KCGMH., Results: In phase 1, embryo abandonment was associated with the production of more mature oocytes and embryos, the freezing of more embryos, young female age, blastocyst transfer, and positive pregnancy results. After adjustment for confounding factors, only positive pregnancy results (adjusted odds ratio [aOR] 4.38, 95% confidence interval [CI] 3.17, 6.04), the freezing of ≥ 2 embryos (aOR 3.68, 95% CI 3.10, 4.38), the production of ≥ 6 embryos (aOR 1.68, 95% CI 1.03, 2.73), and the use blastocyst transfer (aOR 2.46, 95% CI 1.64, 3.69) remained significantly associated with embryo abandonment. The factors associated with embryo abandonment or possible abandonment were similar in phase 2., Conclusion: For elective blastocyst stage transfer and a freezing strategy performed according to the Taiwan National Assisted Reproduction Act, a young female age ≤ 35 with positive pregnancy status due to the original IVF treatment, the production of ≥ 6 embryos, and the cryopreservation of ≥ 2 blastocysts may increase the likelihood of abandoning embryos in the future.

Purpose: Reactive oxygen species (ROS) are thought to play a critical role in the success of IVF. The relationships between oxidative stress parameters in culture media and IVF outcomes have not been extensively investigated. The objective of this study is to examine the relationships between early human embryonic parameters and levels of ROS in culture media., Methods: This prospective study was conducted with 2633 spent culture media collected from patients undergoing conventional IVF (n = 101) and intracytoplasmic sperm injection (ICSI) (n = 60). Both fertilization and early culture were performed in universal IVF medium and G series medium. ROS levels were measured by chemiluminescence assays using luminol as the probe on days 1, 3, and 5 and determined the relationships of ROS levels with zygote condition, embryo quality, and clinical pregnancy rate., Results: ROS levels per embryo in culture media on the corresponding days 1, 3, and 5 showed significant correlations between each pair in the total cohort. Similar results were observed in the IVF and ICSI groups, but day 1 and day 3 ROS levels were significantly higher in culture media of IVF than of ICSI embryos. ROS levels in culture medium were not significantly associated with embryo quality, blastocyst formation, or arrest. ROS levels on day 1 were similar in media of normally fertilized zygotes, unfertilized oocytes, and polyspermic zygotes and were not associated with delayed embryonic development, high fragmentation, blastocyst formation, or arrest after prolonged culture. ROS levels in media were not associated with the likelihood of conception., Conclusions: ROS levels in culture media may not be an effective indicator of embryo selection for IVF.

Background: Many factors from the oocyte/sperm or the process of fertilization may affect the zygote formation. The zygote score (Z-score) describes the quality of a human zygote based on its pronuclear morphology, nucleolar precursor bodies, and alignment of polar bodies, and it can be used in the selection process at the zygote stage for embryo transfer or cryopreservation., Objective: The aim of this retrospective cohort study was to investigate the relationship between different controlled ovarian stimulation (COS) protocols and the zygote score (Z-score) and to assess the feasibility of the Z-score for predicting embryo survival in the GnRH-antagonist (GnRH-ant) protocol., Methods: It is a retrospective, single-center cohort study. A total of 3,826 zygotes with normal fertilization were analyzed from 744 in vitro fertilization /intra-cytoplasmic sperm injection (IVF/ICSI) cycles (long protocol n = 392; GnRH-ant n = 352) between Jan 2010 and April 2014 in the IVF unit of Chang-Gung Memorial Hospital Kaohsiung Medical Center., Results: The Z-score distribution differed significantly between these two protocols. The overall Z-score was poorer for zygotes from GnRH-ant cycles (p<0.05). Univariate and multivariate analyses indicated the type of COS protocol is one of the main determinants of Z-score grading. Our study found good-quality day 3 embryo/blastocyst formation and the cumulative embryo survival rate were correlated with the Z-score but not the COS protocol. With the GnRH-ant protocol, the number of Z1 in the transferred cohort embryos was significantly correlated with the clinical pregnancy rate (r = 0.976; p = 0.024) and live birth rate (r = 0.971; p = 0.029). This correlation was not seen with the long protocol., Conclusions: The Z-score distribution for the GnRH antagonist cycles was poorer than that of the long protocol, but the Z-score system is a valuable parameter for predicting embryo viability in the GnRH-ant protocol, providing a strong correlation with the clinical pregnancy rate and live birth rate., Competing Interests: The authors have declared that no competing interests exist.

Objective: To assess whether progesterone elevation is the only factor in determining outcomes of in vitro fertilization (IVF)., Design: Retrospective cohort study., Setting: Infertility clinic at Kaohsiung Chang Gung Memorial Hospital, Taiwan., Patient(s): One thousand five hundred eight women undergoing a total of 1,508 IVF cycles., Intervention(s): None., Main Outcome Measure(s): Clinical pregnancy and live-birth rates., Result(s): Patients were classified into four subgroups according to their progesterone concentration on the day of human chorionic gonadotropin (hCG) triggering. The clinical pregnancy and live-birth rates were statistically significantly associated with the age of the woman, the day of embryo transfer, the progesterone concentration on the day of hCG administration, the number of transferred embryos, and the number of top-quality embryos transferred. However, after omitting the women with the highest progesterone concentration (≥1.94 ng/mL), only four factors-patient age, day of embryo transfer, number of transferred embryos, and number of top-quality embryos transferred-were statistically significantly associated with the clinical pregnancy and live-birth rates., Conclusion(s): Progesterone concentration on the day of hCG administration is not the only factor determining the clinical pregnancy and live-birth rates. Fresh embryos from women should be frozen with extremely high progesterone concentrations. Each patient's general condition and the capacity for frozen-thawed embryo transfer should be considered before implantation., (Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

Objective: To determine whether or not the level of serum anti-Müllerian hormone (AMH) is related to early ovarian aging in young women (< 35 years of age) undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles., Design: Retrospective cohort study., Setting: An IVF laboratory in a university hospital in Taiwan. Patient (s): 70 young women (< 35 years of age) with low level of serum AMH (< 2 ng/ml) and 104 young women with level of serum AMH (≥ 2 ng/ml) who underwent IVF/ICSI cycles between January 2011 and November 2012 were enrolled. Intervention (s): None. Main outcome measure (s): Number of oocytes, fertilization rate, embryo quality, cycle cancellation rate, clinical pregnancy/abortion rate, and perinatal/infant outcomes., Results: The clinical pregnancy rate per transfer was favorable (low AMH group vs. normal AMH group [47.2% and 47.9%]) for women < 35 years of age, including women with a low serum AMH. Similarly, the live birth rate per transfer (low AMH group vs. normal AMH group [37.7% and 35.4%]) and perinatal outcomes were also comparable between the two groups. A significantly higher cycle cancellation was noted in the low AMH group than the normal AMH group (24.2% vs. 7.6%)., Conclusion: Although early ovarian aging should be taken into consideration for young and infertile women with low AMH level than expected, our results suggest that low serum AMH level may suggest early ovarian aging in accelerated oocyte loss only, but may not fully represent "early ovarian aging" based on the favorable outcomes of pregnancy.

Objective: To compare the differences between the right and left ovarian responses in patients undergoing standard in vitro fertilization (IVF) treatment., Design: Prospective, observational study., Setting: A university hospital in Taiwan., Patient(s): Seven hundred eighty-nine consecutive patients undergoing IVF or intracytoplasmic sperm injection., Intervention(s): None., Main Outcome Measure(s): The number of oocytes retrieved, fertilization rates, number of grade 1 zygotes and number of embryos produced, embryo arrest-free rate, and mean number of top-quality embryos on the day of transfer., Result(s): The number of follicles > or = 16 mm in diameter on day 2.5 versus 2.2 of human chorionic gonadotropin (hCG) administration, the number of follicles > or = 10 mm in diameter on hCG day 6.5 versus 6.1, the number of oocytes (3.6 vs. 3.0), the percentage fertilization (79.9 vs. 74.6%), and the mean number of mature oocytes and grade 1 zygotes (2.3 vs. 1.8 and 1.79 vs. 0.93, respectively), and the mean available number of top-quality embryos on transfer day 1.39 versus 0.73 were all statistically significantly greater in the right compared with the left ovary; however, the pregnancy and implantation rates were similar., Conclusion(s): There are statistically significant differences between the right and left ovarian responses in IVF patients with healthy ovaries, and the right ovarian responses are superior to the left ovarian responses., (Crown Copyright 2010. Published by Elsevier Inc. All rights reserved.)

In unselected IVF patients, zona-free and laser zona-assisted hatching produced a comparable effect on blastocyst transfer.

Objective: To test the safety and efficacy of a modified laser-assisted zonal opening method for human embryo biopsy., Study Design: The embryo was treated with a modified method to create an ample perivitelline space between the zona pellucida and underlying blastomeres. This was done to protect the blastomeres from damage by laser treatment of the zona pellucida. Subsequently, the zona pellucida was completely perforated using a diode laser, and the targeted blastomere was aspirated. In vitro embryo development of the 40 biopsied embryos was compared with that of 322 corresponding control embryos without embryo biopsy., Results: Forty targeted blastomeres were successfully extracted from 40 human embryos. The incidence of embryo development to the blastocyst stage was not different between the biopsied and nonbiopsied groups (55% vs. 54.3%). Furthermore, the percentage of top-scoring blastocysts from biopsied embryos was also similar to that of control embryos (77.3% vs. 78.9%)., Conclusion: These preliminary results demonstrate that the modified technique for human embryo biopsy in preimplantation genetic diagnosis is simple and safe.

Background: This study compares the influence of recombinant (r)FSH and urinary (u)FSH stimulation on oocyte and embryo quality in patients undergoing ICSI., Methods and Results: Denuded oocyte maturity in ICSI patients was graded on a scale from metaphase II (MII) to prophase for nuclear maturation of oocytes. The relationships of cumulus-free oocyte maturational profiles with in vitro outcome parameters were evaluated. In the study population with an unknown distribution of FSH receptor polymorphisms, the ovarian response to rFSH stimulation was significantly different from that of uFSH stimulation, including lower number of oocytes retrieved/oocytes in MII, higher fertilization rates and higher good quality embryo rates. In the study population with a similar distribution of FSH receptor polymorphisms, the ovarian responses to rFSH were lower numbers of oocytes in MII, higher fertilization rates and higher good quality embryo rates, but the total number of oocytes retrieved was not influenced, in comparison with ovarian stimulation with uFSH., Conclusions: rFSH stimulation appears to influence oocyte quality and subsequent embryo quality in comparison with uFSH stimulation. FSH receptor polymorphisms seem to be an intrinsic factor influencing the ovarian response to FSH stimulation.

Objective: To examine the implantation potential of embryos from assisted reproductive technology cycles with low embryo production and to assess the effects of clinical variables and embryo scores (ES) on pregnancy outcome., Design: Prospective clinical study., Setting: Assisted reproductive technology unit in a tertiary medical center., Patient(s): From July 1998 to December 2001, 280 cycles in 229 infertile couples produced a limited number of one, two, or three embryos 3 days after oocyte retrieval and underwent fresh embryo transfer (ET)., Intervention(s): Embryos with two or more blastomeres were scored and transferred., Main Outcome Measure(s): ES and implantation rate per ET., Result(s): Of 863 fresh ET cycles during the study period, 32.4% (280) were low embryo producers. Among them, there were no significant differences in average ES of individual embryos in single, dual, or triple ET or in embryos obtained from patients with low or high E2 responses, or young or old age. Embryos derived from conventional IVF had a better ES than those derived from intracytoplasmic sperm injection. The clinical pregnancy rate was strongly correlated with the cumulative ES. Implantation rates were similar among and between groups, with an average rate of 15.9%., Conclusion(s): Embryos of low embryo producers had an inherently low implantation potential that appeared to be unrelated to the number of embryos transferred, female age, ovarian E2 genesis, or fertilization method. The cumulative ES can serve as a predictor of pregnancy.

Purpose: To date, the impact of oocyte maturity at aspiration on the blastocyst formation in vitro has not been fully evaluated. This study was undertaken to assess the influence of oocyte maturity in patients undergoing in vitro fertilization and blastocyst transfer program., Methods: A total of 1278 oocytes derived from 147-IVF cycles were retrospectivly analyzed. Oocyte maturity was graded on a scale from 1 to 5 based on the morphology of the ooplasm, cumulus mass, corona radiata, and membrana granulosa cells., Results: Mature oocytes yielded the highest fertilization rates. Although the cleavage rates were similar in both groups, the percentage of poor morphology, day-3 embryos from the nonmature-oocyte group was significantly higher than from the mature-oocyte group (54.7% vs. 15.5%, P < 0.001). Although good morphology, day-3 embryos were collected from nonmature oocytes, the incidence of these embryos developing to the blastocyst stage was significantly less than from mature oocytes (33.3% vs. 71.2%, P < 0.001). Although blastocyst stage embryos were collected from nonmature oocytes, the incidence of these embryos developing to the top-scoring blastocysts was significantly less than from mature oocytes (58.3% vs. 89.5%, P < 0.001)., Conclusions: These phenomena suggest that oocyte maturity produced in vivo determine the fertilization potential and subsequent blastocyst quality in vitro.

Background: The purpose of this study was to evaluate the ability of using the Z-score alone, or, in combination with the day 3 embryo morphology score, to predict embryo viability at day 5 from a large cohort of embryos derived from patients undergoing treatment with IVF/ICSI., Methods and Results: In this retrospective study, a total of 1894 zygotes from 346 treatment cycles (295 couples) was analysed between January 2001 and May 2002. The Z-scoring system was useful in predicting day 5 embryo survival. The mean +/- SD day 5 embryo survival rates were 78.2 +/- 1.7, 49.0 +/- 2.5, 21.4 +/- 3.2 and 11.8 +/- 5.6% for Z-1, Z-2, Z-3 and Z-4 zygotes groups respectively. Embryos derived from Z-1 scores and grade I day 3 embryo scores showed the best day 5 embryo survival and a very high implantation potential., Conclusions: These data suggest that a combined evaluation of the Z-score and day 3 embryo morphology is highly predictive of embryo outcome after IVF/ICSI. The Z-score could be of great help in the selection of embryos for cultures extended to later stages. The Z-score alone, or preferably in combination with day 3 embryo morphology, is useful in the determination of the most suitable embryos and the number of embryos for transfer, thus achieving the optimal chance of conception while reducing the risk of high order multiple pregnancy.

Objective: To examine the pregnancy potential of frozen-thawed blastocysts that underwent quarter laser-assisted hatching (AH) at the cleaving stage before freezing and to compare clinical and embryo characteristics between the groups that succeeded in and failed to achieve pregnancy., Design: Prospective observational study., Patient(s): Thirty-four of 112 patients with frozen blastocysts requiring transfer of thawed embryos between January 2000 and December 2001., Setting: Assisted reproductive technology unit in a tertiary medical center., Intervention(s): Embryos of patients undergoing blastocyst transfers routinely underwent quarter AH using a nontouch 1.48-microm diode laser. Blastocysts not transferred were cryopreserved using a six-step freezing protocol with glycerol as the cryoprotectant., Main Outcome Measure(s): Postthaw embryo survival and zona pellucida (ZP) maintenance, implantation rate, and clinical pregnancy rate per transfer of thawed blastocysts., Result(s): A total of 118 frozen blastocysts was thawed. Of these, 89 (75.4%) embryos survived and were transferred in 35 cycles. Assisted hatching-manipulated ZP tolerated the freeze-thaw procedures without shape distortion in surviving embryos. Eleven (31.4%) clinical pregnancies with 15 intrauterine gestational sacs occurred, for an implantation rate of 16.9%. Those who succeeded in pregnancy tended to have more embryos available before freezing, more original blastocysts of good quality for freezing, and more surviving blastocysts after thawing for transfer than did those who failed to achieve pregnancy., Conclusion(s): An acceptable clinical pregnancy rate was obtained from transfer of thawed blastocysts that underwent laser AH on the ZP at the day 3 cleaving embryo stage in fresh cycles in selected patients. Embryo characteristics before freezing played major roles in determining implantation potential of thawed embryos.

Purpose: We investigated whether the human oocyte maturity at the removal of cumulus/corona cells affects the embryo outcome in vitro., Methods: A total of 620 oocytes, which subsequently underwent blastocyst culture, were included in this analysis. Oocytes that were in prophase or Metaphase I of meiosis at the removal of cumulus/corona cells were in Group II. Oocytes that were in Metaphase II at the removal of cumulus/corona cells were in Group I., Results: Group I oocytes yielded the highest fertilization rates (96.3% vs. 77.1%, P < 0.001). The incidence of Group II oocytes developing to the blastocyst stage was significantly less than from Group I oocytes (38.1% vs. 86.1%, P < 0.001). The percentage of top-scoring blastocysts from Group I oocytes was higher than that of Group II oocytes (95.4% vs. 76.2%, P < 0.001)., Conclusions: Oocyte maturity at the removal of cumulus/corona cells needs to be considered in selecting good quality blastocysts for embryo transfer.