P–152 Morphokinetic and maternal profiles of embryos derived from centrally granulated oocytes vary with their ability to implant

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
C Moutier ◽  
A Bartolacci ◽  
D Turchi ◽  
M Lain ◽  
D Pignataro ◽  
...  

Abstract Study question Does oocyte central granularity (CG) impact embryo morphokinetics and does this change with embryo implantation ability and maternal profile? Summary answer Oocyte CG slows fertilization and cleavage morphokinetics in overall derived embryos, but not in those capable to implant, an ability associated with lower maternal age. What is known already Oocyte morphology is easily accessible after pre-ICSI oocyte denudation, but the implications of morphological alterations for oocyte developmental competence are not entirely known. The presence of a centrally located granular area in the ooplasm was previously associated with alterations in the actin cytoskeleton and meiotic spindle, both potentially affecting meiosis completion, fertilization dynamics and embryo mitotic divisions. In fact, we have recently reported lower fertilization rates and delayed pronuclei fading and first cleavage associated with the presence of CG in oocytes subjected to ICSI. Study design, size, duration Retrospective analysis including 1378 control ICSI cycles providing only morphologically normal oocytes (1225 patients) and 220 CG cycles (201 patients) providing normal and CG oocytes, from July 2014 to March 2020. Morphokinetic parameters were compared between embryos from control and CG oocytes, as well as among embryos from control oocytes reaching implantation (C-I) and embryos from CG oocytes achieving (CG-I) or not (CG-NI) implantation. Maternal profiles were compared between CG-I and CG-NI. Participants/materials, setting, methods Oocytes were recovered from patients after controlled ovarian stimulation and ovum pick-up. Following ICSI, embryo culture was performed in a time-lapse incubator with annotation of time of pronuclei fading (tPNf) and cleavage times t2, t3, t4, t5 and t8. Morphokinetic data were retrospectively coupled with implantation outcomes of single transfers and of double transfers achieving double or no implantation. Differences were assessed with Chi-square and Kruskal Wallis tests. Main results and the role of chance Patients providing CG oocytes (n = 201) presented higher maternal age (37.4 ± 4.4 vs. 36.7 ± 4.3; p = 0.005), higher basal FSH (8.52 ± 3.7 vs. 7.62 ± 2.8 IU/L; p = 0.002) and lower AMH levels (2.2 ± 2.2 vs. 2.9 ± 3.1 ng/mL; p < 0.001) compared to control patients (n = 1225). Morphokinetic parameters from tPNf to t4 were faster in embryos derived from oocytes with normal morphology (control; n = 6947) compared to embryos derived from CG oocytes (n = 382; tPNf: 24.0 ± 3.8 vs. 24.6 ± 3.6; t2: 27.0 ± 4.3 vs. 27.6 ± 4.1; t3: 37.0 ± 5.7 vs. 37.4 ± 5.7; t4: 39.4 ± 6.4 vs. 40.1 ± 6.2 hours; p < 0.05). In addition, CG-NI (n = 103) embryos were slower than CG-I (n = 13) and C-I (n = 226) embryos for tPNf, t2, t3, t4 and t8 (p < 0.05), while CG-I did not differ from C-I embryos (p > 0.05; tPNf: 22.4 ± 2.6 vs. 22.0 ± 2.5; t2: 24.9 ± 2.7 vs. 24.6 ± 2.8; t3: 36.0 ± 3.5 vs. 35.4 ± 3.0; t4: 36.7 ± 3.5 vs. 36.7 ± 3.6 hours, for C-I and CG-I, respectively). Finally, patients providing CG-I embryos (n = 10) were younger than those providing CG-NI embryos (n = 65; 31.3 ± 4.6 vs. 38.0 ± 3.9; p < 0.05). Limitations, reasons for caution Our study is subjected to the intrinsic limitations of a retrospective analysis, the results presented could have been affected by variables that are uncontrolled for. Other studies are necessary to assess the impact of CG on clinical outcomes. Wider implications of the findings: The findings indicate that early developmental morphokinetics and maternal age constitute valid parameters for the decision of whether to transfer CG-derived embryos, as well as for the transfer prognosis. Trial registration number Not applicable

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
J Buratini ◽  
G Sivelli ◽  
P Novara ◽  
F Brambillasca ◽  
L Mura ◽  
...  

Abstract Study question Do maternal AMH and basal FSH profiles impact embryo morphokinetics and does this relationship change with maternal age? Summary answer Embryo morphokinetics varies with basal FSH and AMH levels and this relationship changes in advanced maternal age (AMA). What is known already Basal FSH and AMH levels have been utilised as markers of ovarian reserve/response and IVF/ICSI outcomes. Basal FSH better reflects post-IVF/ICSI live birth occurrence in pre-AMA patients, while AMH appears more robust in AMA patients. Whilst plasma AMH levels reflect oocyte yield, recent data suggest that plasma basal FSH and intrafollicular AMH levels specifically reflect oocyte quality. Oocyte and embryo developmental competence is associated with faster fertilisation and embryonic morphokinetics. The assessment of the relationship between developmental morphokinetics and maternal basal FSH and AMH plasma profiles shall contribute for a better understanding of their roles as fertility markers and regulators. Study design, size, duration Retrospective cohort study including 1961 first autologous ICSI cycles performed from 2014 to 2020, providing 10774 embryos grouped according to maternal AMH and basal FSH levels in: CF [concordant favourable; AMH>1 (ng/mL), FSH≤10 (IU/L); n = 8055)]; DFA (discordant with favourable AMH; AMH>1, FSH>10; n = 768); DFF (discordant with favourable FSH; AMH≤1, FSH≤10; n = 1362) and CU (concordant unfavourable; AMH≤1, FSH>10; n = 589). Morphokinetic parameters were compared among groups in total, ≤35 (pre-AMA) and >35 (AMA) years old patients, separately. Participants/materials, setting, methods Patients aged 20 to 45 with FSH and AMH levels measured on menstrual cycle day 2 underwent ovarian stimulation, ovum pick-up and ICSI. Embryos were cultured in a time-lapse incubator (Embryoscope). Fertilisation and cleavage morphokinetic parameters [tPNa (time of pronuclei appearance, tPNf (time of pronuclei fading), t2, t3, t4, t5 and t8] were annotated and compared among AMH/FSH groups with the Kruskal-Wallis nonparametric test, followed by a post hoc multiple comparison with Bonferroni correction. Main results and the role of chance In overall embryos, tPNa, tPNf, t2 and t3 varied between AMH/FSH groups. CF were faster than CU embryos for all these parameters [(mean±SEM; hours); tPNa: 6.8±0.02 vs. 7.1±0.08; tPNf: 24.13±0.04 vs. 24.49±0.14; t2: 27.05±0.05 vs. 27.36±0.16; t3: 36.94±0.07 vs. 37.54±0.22 (p < 0.001)]. In addition, CF were faster than DFA (p < 0.001), but not than DFF embryos, for tPNf (CF: 24.13±0.04; DFF: 24.23±0.14; DFA: 24.57±0.14) and t2 (CF: 27.05±0.05; DFF: 27.22±0.12, DFA: 27.42±0.15). In AMA patients, faster morphokinetics was observed when one or both hormonal values were favourable; tPNf, t2 and t3 were reached earlier in CF compared to CU (tPNf: 24.17±0.05 vs. 24.52±0.15; t2: 27.13±0.06 vs. 27.43±0.17; t3: 37.09±0.08 vs. 37.63±0.24; p < 0.05), but not to DFF (tPNf: 24.22±0.12; t2: 27.27±0.14, t3: 36.92±0.19) or DFA embryos (tPNf: 24.39±0.14; t2: 27.34±0.17, t3: 37.26±0.23). Differently, in pre-AMA patients, faster morphokinetics was associated with favourable basal FSH regardless of AMH levels; tPNa and tPNf were reached earlier in CF compared to DFA and CU (p < 0.005), but not to DFF embryos (tPNA: 6.68±0.03, 6.96±0.13, 7.13±0.14, 7.12±0.19; tPNf: 24.05±0.07, 24.27±0.21, 25.14±0.34, 24.37±0.36; for CF, DFF, DFA and CU, respectively). Limitations, reasons for caution Our study is subjected to the intrinsic limitations of a retrospective analysis and the results could have been affected by variables that were not controlled for. Wider implications of the findings: The findings suggest that lower basal FSH levels are associated with faster early morphokinetics likely reflecting superior oocyte quality in pre-AMA patients. The present data may contribute to improve ART prognostic strategies and provide valuable clues for a better understanding of hormonal regulation of oocyte developmental competence. Trial registration number Not applicable


2015 ◽  
Vol 104 (5) ◽  
pp. 1175-1181.e2 ◽  
Author(s):  
Daniel Bodri ◽  
Takeshi Sugimoto ◽  
Jazmina Yao Serna ◽  
Masae Kondo ◽  
Ryutaro Kato ◽  
...  

2015 ◽  
Vol 27 (1) ◽  
pp. 210
Author(s):  
M. Taniai ◽  
M. Takayama ◽  
O. Dochi ◽  
K. Imai

Bovine IVF embryos are evaluated morphologically using light microscopy just before transfer. However, this evaluation method is subjective, and an objective method with more certainty is needed. Sugimura et al. (PLoS ONE 2012 7, e36627) reported a promising system for selecting healthy IVF bovine embryo by using time-lapse cinematography and 5 prognostic factors. This study was to investigate the efficacy of a 2-step evaluation system of IVF embryos using microscopy for selecting high developmental competence IVF embryos. Cumulus-oocyte complexes (COC) were collected by ovarian follicular aspiration (2 to 5 mm diameter) obtained from a local abattoir. The COC (n = 488) were matured in TCM-199 medium supplemented with 5% calf serum (CS) and 0.02 IU mL–1 of FSH at 38.5°C for 20 h in an atmosphere of 5% CO2 (20 COC 100 µL–1 droplets). After 10 h of gametes co-culture (5.0 × 106 sperm cells mL–1), the presumptive zygotes were cultured in 125 µL of CR1 aa medium supplemented with 5% CS in well of-the-well culture dishes (AS ONE, Japan; 25 zygotes well–1) at 38.5°C in an atmosphere of 5% CO2, 5% O2, and 90% N2 for 9 days. Two-step evaluations of embryos were done at 27 and 55 h post-IVF (hpi). In the first step of evaluation, cleavage patterns at 27 hpi were categorized as mono-cell, 2-cell with even blastomeres and without fragments (normal cleavage), 2-cell with uneven blastomeres, and ≥3 blastomeres. During the second step of evaluation, embryos were classified by their number of blastomeres (2 to 5 cells, 6 to 8 cells, and >8 cells) and the absence or presence of multiple fragments (<20 or >20%) at 55 hpi. The data were analysed by chi-square test. The blastocyst rate (BL%) of embryos cleaved before 27 hpi (56.6%, n = 106) was higher (P < 0.01) than those of embryos cleaved after 27 hpi (37.0%, n = 235). A greater percentage (P < 0.05) of 2-cell embryos with normal cleavage (68.0%, n = 50) developed to blastocysts than from with =3 blastomeres at 27 hpi (40.6%, n = 32). Superior BL% (P < 0.01) was obtained from embryos categorized as 6- to 8-cell stage (58.6%, n = 140) and >8 cell stage (70.6%, n = 25) compared with those embryos at the 2- to 5-cell stage at 55 hpi (26.1%, n = 176). Embryos with no fragments (58.0%, n = 467) had higher BL% (P < 0.01) compared with those with <20% fragments (30.7%, n = 127) and having with >20% fragments (17.5%, n = 25) at 55 hpi. The highest of BL% was observed in embryos showing a normal cleavage to 2-cells with at 27 hpi and having >6 cells with no fragments at 55 hpi (95.2%, n = 21, P < 0.01). These results demonstrate that the 2-step evaluation system at 27 and 55 hpi using microscopy is an effective method for selecting IVF embryos with high developmental competence.


Zygote ◽  
2021 ◽  
pp. 1-8
Author(s):  
Alessandro Bartolacci ◽  
Mariabeatrice Dal Canto ◽  
Maria Cristina Guglielmo ◽  
Laura Mura ◽  
Claudio Brigante ◽  
...  

Abstract Given the importance of embryo developmental competence assessment in reproductive medicine and biology, the aim of this study was to compare the performance of fertilization and cleavage morphokinetics with embryo morphology to predict post-ICSI live birth. Data from embryos cultured in a time-lapse microscopy (TLM) incubator and with known live birth outcomes (LB: embryos achieving live birth, n = 168; NLB: embryos not achieving live birth, n = 1633) were used to generate receiver operating characteristic (ROC) curves based on morphokinetic or morphological scores, and the respective areas under the curve (AUC) were compared. The association between live birth and 12 combinations of four morphokinetic quality degrees (A–D) with three morphological quality degrees (A–C) was assessed using multivariate analysis. Morphokinetic parameters from tPNa to t8 were reached earlier in LB compared with NLB embryos. The ROC curve analysis indicated that morphokinetic information is more accurate than conventional morphology to predict live birth [AUC = 0.64 (95% CI 0.58–0.70) versus AUC = 0.58 (95% CI 0.51–0.65)]. The multivariate analysis was in line with AUCs, revealing that embryos with poor morphokinetics, independently of their morphology, provide lower live birth rates (P < 0.001). A considerable percentage of embryos with top morphology presented poor morphokinetics (20.10%), accompanied by a severely reduced live birth rate in comparison with embryos with top morphology and morphokinetics (P < 0.001). In conclusion, TLM-derived early morphokinetic parameters were more predictive of live-birth achievement following ICSI than conventional morphology.


2015 ◽  
Vol 27 (1) ◽  
pp. 208
Author(s):  
S. Matoba ◽  
T. Somfai ◽  
T. Nagai ◽  
M. Geshi

Previously, an early first cleavage and a second cleavage after IVF with a normal cleavage pattern defined by even blastomeres without fragments or protrusions was found to be a potent marker for the selection of embryos with high developmental competence (Sugimura et al. 2012 PLoS ONE 7, e36627). The aim of this study was to investigate the effects of bulls and X-sorting of sperm on the ability of these simple noninvasive markers to predict the potency of bovine IVF embryos to develop to the blastocyst stage in vitro. Immature oocytes were matured in TCM199 supplemented with 0.02 armour unit mL–1 FSH and 5% calf serum at 38.5°C in 5% CO2 and 95% air for 22 to 23 h. After maturation, oocytes were inseminated with either of non-sorted frozen-thawed sperm from 3 bulls (A–C) or X-sorted sperm of bull A. Putative zygotes were cultured (IVC) in CR1aa medium supplemented with 5% calf serum and 0.25 mg mL–1 linoleic acid albumin at 38.5°C in 5% CO2, 5% O2, and 90% N2 for 216 h. Embryo kinetics were observed individually by time-lapse cinematography (CCM-1.3Z; Astec, Fukuoka, Japan; Sugimura et al. 2010 Biol. Reprod. 83, 970–978). First and second cleavage kinetics and pattern were categorized according to Sugimura et al. (2012). For each bull, blastocyst development from embryos possessing the following 3 selection markers was compared: (marker 1) the first cleavage within 28 h after IVF, (marker 2) marker 1 combined with 2 even blastomeres without fragments or protrusions, and (marker 3) marker 2 combined with the second cleavage within 50 h after IVF with ≥6 even blastomeres without fragments or protrusions, respectively. Data were analysed by the Yates' corrected chi-square test. A total of 823 oocytes were used in at least 3 replications. When non-sorted sperm was used for IVF, there was not difference (P > 0.05) in total blastocyst formation rates on Day 8 (Day 0 = IVF) among bulls (ranging between 49.5 and 60.8%); however, blastocyst formation rate of embryos generated from X-sorted sperm of bull A (39.5%) was lower (P < 0.05) compared with other groups despite of similar cleavage rates. Embryos having marker 3 criteria developed to the blastocysts stage at significantly higher rates than those having marker 1 criteria in case of non-sorted sperm of bulls A, B, C, and X-sorted sperm of bull A (75.9, 87.0, 90.0, and 75.0% v. 59.5, 62.2, 63.6, and 46.3%, respectively). In groups produced from non-sorted sperm of bulls A, B, C, and X-sorted sperm of bull A, blastocyst development rates of embryos with marker 2 criteria (73.7, 75.0, 90.0, and 65.8%, respectively) were higher (P < 0.05) than those of embryos having marker 1 criteria but did not differ significantly from those with marker 3 criteria. Our results reveal that a first cleavage within 28 h after IVF to 2 even blastomeres without fragments or protrusions are potent predictive markers of the developmental competence of bovine embryos to the blastocyst stage regardless of bulls and sperm sorting.Research was partly supported by JSPS KAKENHI (26450388).


2020 ◽  
Vol 35 (1) ◽  
pp. 24-31 ◽  
Author(s):  
C R Sacha ◽  
I Dimitriadis ◽  
G Christou ◽  
K James ◽  
M L Brock ◽  
...  

Abstract STUDY QUESTION Is there an effect of male factor infertility (MFI) on either early or late morphokinetic parameters obtained during embryonic culture to blastocyst stage in a time-lapse imaging (TLI) incubator? SUMMARY ANSWER Neither mild nor severe MFI had an impact on overall time to blastocyst or duration of individual cleavage stages in the total embryo population. WHAT IS KNOWN ALREADY Prior studies have suggested that paternal DNA and sperm quality affect embryo morphokinetic parameters, but the impact of MFI is not fully understood. STUDY DESIGN, SIZE, DURATION This retrospective cohort study, at a major academic fertility centre, included 536 couples (women, ≤44 years of age) undergoing IVF between September 2013 and September 2016. PARTICIPANTS/MATERIALS, SETTING, METHODS Data from 4126 embryos cultured to the blastocyst stage in a TLI-monitored incubator were retrospectively reviewed. Embryos derived from the sperm of men with MFI were compared with those derived from patients with other infertility diagnoses. Generalized fixed and random effects models, t-test and χ2 were used as appropriate. MAIN RESULTS AND THE ROLE OF CHANCE Couples with MFI had a higher rate of ICSI utilization and fewer usable embryos on average, and the men were older compared with couples with other diagnoses. Additionally, the women in MFI couples were younger and had higher antral follicle counts (AFCs) and higher anti-Müllerian hormone (AMH) levels compared with the other women undergoing IVF. When controlling for maternal and paternal ages, AMH and fertilization method (conventional IVF versus ICSI), neither mild nor severe MFI affected duration of individual cleavage stages or overall time to the blastocyst stage, when all or only usable embryos were examined (coefficient 0.44 hours in all embryos, P = 0.57; coefficient 0.39 hours in usable embryos, P = 0.60). Whether the sperm was surgically extracted similarly had no significant effect on embryo morphokinetic parameters. When the fertilization method was assessed independently, ICSI lengthened the overall time to blastocyst stage by 1.66 hours (P = 0.03) on average, primarily due to an increase in duration of the time from 5-cell embryo stage to early blastulation (P5SB). LIMITATIONS, REASONS FOR CAUTION This large cohort study avoided embryo selection bias due to random assignment of embryos to the TLI incubators. However, our findings may not be generalizable to groups under-represented in our clinic population. Future studies should also evaluate the impact of male hormonal status and detailed sperm morphology, such as head versus flagellum defects, on embryo morphokinetic development. WIDER IMPLICATIONS OF THE FINDINGS Our findings suggest that the fertilization method rather than MFI per se impacts time to early blastulation. The clinical implications of this effect on embryo development warrant further investigation. STUDY FUNDING/COMPETING INTEREST(S) There were no sources of funding for this study. There are no competing interests. TRIAL REGISTRATION NUMBER N/A


1996 ◽  
Vol 11 (S2) ◽  
pp. S27-S27
Author(s):  
Sharon E. Mace ◽  
William Kriegsman

Hypothesis: Do paramedics influence the outcome of out of hospital cardiac arrests in a rural setting?Methods: Retrospective analysis of cardio-respiratory arrests of ALS-EMS system in rural Southeastern Alaska for 9 years. There were two patient groups treated by EMT-III or paramedics. EMT-III vs. paramedics differ in training/experience but not technical skills. Statistical analyses were done by chi square.Results: Thirty-seven patients (52%) were treated by paramedics, thirty-four (48%) by EMT-III. Demographics/CPR variables for the groups were not significantly different. Comparing paramedics vs. EMT-III: successful ET placement (87% vs. 62%, p <0.02), successful IV placement (87% vs. 62%, p <0.02), return of spontaneous circulation (ROSC) 46% (17/37 pts.) vs. 18% (6/34 pts.) (p = 0.01), ICU admission 38% (14/37) vs. 15% (5/34) (p = 0.03), hospital discharge 20% (7/35) vs. 9% (3/34) (p = NS). There was no correlation between successful ET placement or IV insertion and outcome.


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