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OBJECTIVE: To study whether severe male factor infertility (SMF), reflected by oligozoospermia, impacts embryo morphokinetic behavior in low-prognosis women as stratified by the Patient-Oriented Strategies Encompassing IndividualizeD Oocyte Number (POSEIDON) criteria. DESIGN: Cohort study. SETTING: Private university-affiliated in vitro fertilization center. PATIENT(S): A total of 10,366 injected oocytes from 2,272 women who underwent intracytoplasmic sperm injection cycles between March 2019 and April 2022. INTERVENTION(S): Patients were divided into 8 groups according to the POSEIDON criteria (1-4) and the presence or absence of SMF. A control group of normoresponder patients was included. Kinetic markers from the point of insemination were recorded in the EmbryoScope incubator. MAIN OUTCOME MEASURE(S): Morphokinetic milestones and intracytoplasmic sperm injection clinical outcomes. RESULT(S): Embryos from patients in the POSEIDON 1 group showed significantly slower timing to pronuclear appearance, timing to pronuclear fading (tPNf), timing to 2 (t2), 3 (t3), 4 (t4), 6 (t6), and 7 (t7) cells than those from the control group. Known Implantation Diagnosis Score ranking was significantly different between the SMF and non-SMF (nSMF) subgroups in both POSEIDON 1 as well as control groups. Embryos from patients in the POSEIDON 2 group showed significantly slower timing to pronuclear appearance, t4, t6, t7, timing to 8 cells (t8), and timing to morulae than those from the control group. Embryos in the POSEIDON 2 SMF subgroup took longer than those in the POSEIDON 2 nSMF subgroup and those in both control subgroups to achieve tPNf, t2, t3, timing to 5 cells (t5), timing to start blastulation, and timing to blastulation. Known Implantation Diagnosis Score ranking was significantly different between the SMF and nSMF subgroups in both POSEIDON 2 as well as control groups. Embryos from patients in the POSEIDON 3 group showed significantly slower t8 and duration of the second cell cycle (t3-t2) than those from the control group. Known Implantation Diagnosis Score ranking was significantly different across the subgroups. Embryos derived from patients in the POSEIDON 4 group showed significantly slower tPNf, t2, t3, t4, t5, t6, t7, t8, timing to complete t4-t3 synchronous divisions, and timing to complete t8-t5 synchronous divisions than those from the control group. Known Implantation Diagnosis Score ranking was significantly different between the SMF and nSMF subgroups in both POSEIDON 4 as well as control groups. Irrespective of sperm quality, clinical outcomes significantly improved in the control subgroups compared with those in the POSEIDON 2 and 4 subgroups. CONCLUSION(S): Embryos in the SMF groups presented lower Known Implantation Diagnosis Score ranking than those in the nSMF groups in both POSEIDON 1-4 and control groups, suggesting that cumulative differences result in worse morphokinetic development when the algorithm is used.

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The goal for the present study was to investigate the effect of aneuploidy on embryo morphokinetics events in a time-lapse imaging (TLI) system incubator. This retrospective cohort study was performed in a private university-affiliated in vitro fertilization center, between 2019 March and December 2020. Kinetic data were analyzed in 935 embryos, derived from 316 patients undergoing intracytoplasmic sperm injection cycle with preimplantation genetic testing (PGT) for aneuploidy, individually cultured in a TLI incubator until Day 5 of development. Timing of morphokinetic variables, the incidences of multinucleation, and Known Implantation Data Score (KIDScore)-Day 5 were compared between euploid (n = 352) and aneuploid embryos (n = 583). Aneuploid embryos showed significantly longer timing to complete specific morphokinetic parameters compared to euploidy embryos. Euploidy embryos also showed a significantly higher KIDScore when compared with the aneuploidy ones. Our evidence suggests that TLI monitoring may be an adjunct approach to select embryos for PGT; however, cautious investigation is still needed.

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Supplementation of culture media with IGF-1 during in vitro culture of embryos has had controversial results over the years. In the present study, we show that differences previously observed in response to IGF addition might be related to intrinsic heterogeneity of the embryos. In other words, the effects exerted by IGF-1 are dependent on the characteristics of the embryos and their ability to modulate metabolism and overcome stressful conditions, such as the ones found in a non-optimized in vitro culture system. To test this hypothesis, in vitro produced bovine embryos with distinct morphokinetics (fast- and slow-cleavage) were submitted to treatment with IGF-1 and then evaluated for embryo production rates, total cell number, gene expression and lipid profile. Our results show that remarkable differences were found when fast and slow embryos treated with IGF-1 were compared. Fast embryos respond by upregulating genes related to mitochondrial function, stress response, and lipid metabolism, whereas slow embryos presented lower mitochondrial efficiency and lipid accumulation. We conclude that indeed the treatment with IGF-1 selectively affects embryonic metabolism according to early morphokinetics phenotypes, and this information is relevant for decision-making in the design of more appropriate in vitro culture systems.

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The goal for the present study was to investigate whether previous infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) may compromise embryo morphokinetics and implantation. For that, a historical cohort study was performed in a private university-affiliated in vitro fertilization center. The study included 1628 embryos from 88 patients undergoing intracytoplasmic sperm injection (ICSI) cycles. Patients were age-matched in a 1:3 ratio to either a coronavirus disease (COVID) group, including patients with a positive SARS-CoV-2 immunoglobulin test (n = 22 patients, 386 embryos), or a control group, including patients with a negative SARS-CoV-2 immunoglobulin test (n = 66, 1242 embryos). The effect of previous infection with SARS-CoV-2 on morphokinetic events and ICSI outcomes was evaluated. Embryos derived from patients in the COVID group presented longer time to pronuclei appearance and fading, time to form two, three, four and five cells, and time to blastulation. The durations of the third cell cycle and to time to complete synchronous divisions were also significantly increased in the COVID group compared with the control group, whereas known implantation diagnosis score Day 5 ranked significantly lower in the COVID group. No differences were observed between the COVID and control groups on clinical outcomes. In conclusion, patients planning parenthood, who have recovered from COVID-19 infection, must be aware of a possible effect of the infection on embryo development potential.

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With the growing understanding of in vitro fertilization and reproductive technology, the magnitude of studies related to embryonic evolution has also increased. The optimization of embryo selection is crucial to minimize the risk of multiple pregnancies and to guarantee successful implantation and pregnancy. On the second day of culture, the four-cell embryo can be shaped into different arrangements, such as tetrahedral and planar. Previous studies have shown that mammalian embryos have a tetrahedral shape and that any deviation from this ideal configuration can negatively affect blastocyst development. A few studies have also found that planar embryos would be linked to negative predictors of success for reaching the blastocyst stage and its good quality. Therefore, it seems that tetrahedral should be preferred over planar-shaped embryos for embryonic transfers, but there is still little understanding and evidence about this subject. Thus, the objective of the present paper was to review the available literature on study tendencies to compare tetrahedral and planar-shaped embryos considering their effect on implantation and pregnancy results.

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The objective of this study was to investigate the impact of male age, semen quality and days of ejaculatory abstinence on embryo morphokinetics. A total of 1,220 zygotes obtained from 139 couples in a private in vitro fertilisation centre were analysed. The timing of specific events from the point of insemination, such as timings to pronuclei appearance and fading, to two, three, four, five, six, seven and eight cells and to blastulation were recorded. Multivariate linear regression analysis was used to evaluate the influence of paternal factors on embryo morphokinetic events. Paternal age was positively correlated with delayed cell cleavage and blastulation, and negatively associated with implantation rate, and clinical pregnancy and live-birth chances. The ejaculatory abstinence was inversely correlated with the implantation rate. Inverse relationships were observed between semen parameters (sperm count, progressive sperm motility, total motile sperm count and morphology) and the timing of specific events during embryo development. Sperm morphology was also positively associated with implantation rate and pregnancy and live-birth chances. Increased paternal age and ejaculatory abstinence, and poor semen quality correlate with delayed cell cleavage and blastulation and negatively impact intracytoplasmic sperm injection outcomes.

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OBJECTIVES: To evaluate the association between morphology grading and morphokinetic parameters in blastocyst stage embryos cultured in a time-lapse system. METHODS: This retrospective cohort study included patients offered fertility treatment with autologous oocytes in our clinic between October 2017 and May 2019 using a time-lapse system. The embryos were morphologically graded according to the criteria developed by Gardner and Schoolcraft and their morphokinetic parameters were recorded. RESULTS: Our results indicated that the time of pronuclei fading (tPNf), time to cleavage into two (t2), four (t4), and eight (t8) cells, and time to start of blastulation (tB) were significantly different according to the morphological quality of the blastocysts formed. In the early development stage, tPNf, t2 and t4 differed between good (AA, AB, BA, BB) and poor (CC) quality potential blastocysts. The 8-cell stage time separated embryos graded as AA blastocysts in terms of morphology from embryos graded as BB. Earlier tB correlated with higher quality embryos (AA, AB, BA). CONCLUSION: Our results showed that the first kinetic parameters (tPNf, t2, and t4) distinguished top-graded from low-graded blastocysts. Between top-graded blastocysts, t8 separated BB blastocysts from AA blastocysts. And finally, tB also told apart BB blastocysts from AA, AB, and BA blastocysts. These time-related parameters may be applied even in centers without time-lapse systems.

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OBJECTIVE: Our primary aim was to compare the morphology and morphokinetics on inter- and intra-observer agreement for blastocyst with known implantation outcome. Our secondary aim was to validate the morphokinetic parameters' ability to predict pregnancy using a previous published selection algorithm, and to compare this to standard morphology assessments. METHODS: Two embryologists made independent blinded annotations on two occasions using time-lapse images and morphology evaluations using the Gardner Schoolcraft criteria of 99 blastocysts with known implantation outcome. Inter- and intra-observer agreement was calculated and compared using the two methods. The embryos were grouped based on their morphological score, and on their morphokinetic class using a previous published selection algorithm. The implantation rates for each group was calculated and compared. RESULTS: There was moderate agreement for morphology, with agreement on the same embryo score in 55 of 99 cases. The highest agreement rate was found for expansion grade, followed by trophectoderm and inner cell mass. Correlation with pregnancy was inconclusive. For morphokinetics, almost perfect agreement was found for early and late embryo development events, and strong agreement for day-2 and day-3 events. When applying the selection algorithm, the embryo distributions were uneven, and correlation to pregnancy was inconclusive. CONCLUSIONS: Time-lapse annotation is consistent and accurate, but our external validation of a previously published selection algorithm was unsuccessful.

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OBJECTIVE: To validate a morphokinetic implantation model developed for EmbryoScope on embryos with known outcome, compared to standard morphology in a retrospective single center study. METHODS: Morphokinetic annotation of 768 embryos with known outcome between 2013 -2015; corresponding to 116 D3 fresh embryos, 80 D6 frozen blastocysts, and 572 D5 blastocysts, fresh or frozen. The embryos were ranked by the KIDScore into five classes, KID1-5, and grouped into four classes based on standard morphology. Pregnancy rates, clinical pregnancy rates and live birth rates were compared. Combinations of morphology and morphokinetics were evaluated for implantation rates and live births. RESULTS: Live birth rate increased with increasing KIDScore, from 19% for KID1 to 42% for KID5. Of all live births, KID5 contributed with 71%, KID4 with 20%, KID3 with 4%, KID2 with 4%, and KID1 with 2%. For morphology, the corresponding figure was 43% for Top Quality, 47% for Good Quality, 4% for Poor Quality, and 5% for Slow embryos. For day 3 embryos, KID5 embryos had the highest live birth rates, and contributed to 83% of the live births; whereas the second best morphological class had the highest live birth rate and contributed to most of the live births. For blastocysts, the KIDScore and morphology performed equally well. Combining morphology and morphokinetics indicated stronger predictive power for morphokinetics. CONCLUSIONS: Overall, the KIDScore correlates with both implantation and live birth in our clinical setting. Compared to morphology, the KIDScore was superior for day 3 embryos, and equally good for blastocysts at predicting live births.

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OBJECTIVE: This study aimed to correlate male age with embryo morphokinetic parameters on D3 considering the timing and the exact moment of embryo cleavage. METHODS: Time-lapse imaging was used to produce an ideal cleavage curve for the embryos analyzed. The percentage of embryos under the curve was analyzed and correlated with male age. RESULTS: 32.6% of the embryos from patients aged 28-33 years were under the curve; 36.2% of the embryos from patients aged 34-39 years were under the curve; 41.3% of the embryos from patients aged 40-45 years were under the curve; and 26.3% of the embryos fro patients aged 46-57 years were under the curve. CONCLUSIONS: a statistically non-significant decrease was observed in the percentage of embryos under the optimal cleavage curve on D3 in the group of men aged between 40 and 45 years. Further studies looking into embryos in the blastocyst stage (D5 or D6) are required.