L. Alegre

Analysis of the morphological dynamics of blastocysts after vitrification/warming: defining new predictive variables of implantation

OBJECTIVEnTo describe the morphological dynamics of vitrified/warmed blastocysts and to identify quantitative morphological variables related to implantation. Subsequently, by using the most predictive parameters, to develop a hierarchical model by subdividing vitrified/warmed blastocysts into categories with different implantation potentials.nnnDESIGNnObservational, retrospective, cohort study.nnnSETTINGnUniversity-affiliated private IVF center.nnnPATIENT(S)nThe study included 429 vitrified/warmed blastocysts with known implantation data, which were evaluated by time-lapse imaging. Blastocysts were routinely placed in EmbryoScope (Vitrolife) immediately after warming until transfer.nnnINTERVENTION(S)nNone.nnnMAIN OUTCOME MEASURE(S)nEmbryos were vitrified and warmed by the Cryotop method (KitazatoBiopharma). The studied variables included the initial and minimum thicknesses of zona pellucida (μm), the initial and maximum areas (μm2), the area of inner cell mass (μm2), expansion (whether the embryo reexpands or not after warming), and collapsing or contraction after warming. After defining the optimal ranges according to the consecutive quartiles with the highest probability of implantation, a logistic regression analysis was performed by combining the former variables and the blastocyst morphological classification criteria defined by the Spanish Association of Embryologists into A, B, C, or D categories.nnnRESULT(S)nReexpansion of vitrified/warmed blastocysts correlated strongly with implantation (44.6% for reexpanded vs. 6.5% for the blastocysts that did not reexpand after warming). Throughout the logistic regression analysis, the model identified the maximum blastocyst area, odds ratio (OR) = 0.41 (95% confidence interval [CI], 0.22-0.77), followed by the initial area, OR = 0.62 (95% CI, 0.35-1.08) as the most predictive variables related to implanting embryos. Blastocyst morphology was not considered relevant in our model. The hierarchical tree model subdivided embryos into four categories, A-D, with lowering expected implantation potentials (from 47.3% for A to 14.2% for D).nnnCONCLUSION(S)nThe analysis of warmed blastocysts by time-lapse imaging may provide objective quantitative markers for the blastocyst implantation potential. We propose a hierarchical model to classify vitrified/warmed blastocysts according to their implantation probability. The observed correlations and the proposed algorithm should be validated in a prospective trial to evaluate its efficacy.

Trophectoderm cell cycle length associated with the implantation potential: description of novel embryonic parameters through time-lapse technology

OBJECTIVE: Time-lapse technology allows timing of the most relevant events in embryo development. This increases the probability of selecting the highest quality embryo. However, not all seemingly good quality embryos lead to implantation raising the need of defining new selection variables. A precise morphokinetic evaluation was conducted with the high optical quality incubator Embryoscope Plus_. In order not only improve the embryo selection parameters, but also to introduce new variables, which have not been evaluated so far.

Improvement in reproductive outcome through artificial oocyte activation on fertilization failure cases: a cohort study

MATERIALS AND METHODS: We described the outcome from 273 oocytes from 66 patients who underwent first attempt of ICSI without AOA, TABLE 1. Outcomes by insemination method ICSI IVF P-value High quality total blastulation ratey Mean (SD) (n[702 cycles) 28% (28) (n[381 cycles) 37% (32) <0.05 High quality total blastulation rate -Split groupy Mean (SD) (n[94 cycles) 43% (29) (n[94 cycles) 49% (31) <0.05 First cycles only ICSI IVF P-value High quality total blastulation rate y Mean (SD) (n[365 cycles) 32% (29) (n[251 cycles ) 43% (32) <0.05 High quality total blastulation rate Split-group y Mean (SD) (n[78 cycles) 44% (32) (n[78 cycles) 50% (33) <0.05 y(# day 5 High quality blast + # day 6 High quality blast)y (# 2pn # trans on day 3) FERTILITY & STERILITY_ e221 getting either fertilization failure or low percentage of fertilization (<30%) and were compared with 620 oocytes from the same cohort of patients (84 cycles) in which a new attempt (48 with one cycle and 18 patients with two cycles) were performed with AOA. Study period included between April 2013 and September 2016. The injection of the oocytes by AOAwas carried out by injecting the spermatozoa together with a previous phase of buffered media with Ica (1/3 of pipette in 40x magnification microscope). Later, they were kept for ten minutes in the incubator with fertilization media and ICa in a 37 _ C, 6%CO2 atmosphere. Embryo culture was carried out in standard incubator under culture conditions of 37_C, 6% CO2, 5% O2 atmosphere. Fertilization, pregnancy, implantation and abortion rates were analysed and compared in both groups by X2, t-Student and ANOVA tests when were needed.

Humid vs. dry embryo culture conditions on embryo development: a continuous embryo monitoring assessment

MATERIALS AND METHODS: Embryos were cultured in the time-lapse incubator Geri_ (Genea Biomedx, Australia) with 6 patient-individual chambers. 3 chambers worked under humid conditions (HC) and 3 under dry conditions (DC), 83 and 93 patients respectively. Geri dishes with 80 mL of culture media were covered with an oil overlay, and cultured for 5-6 days. The effects of humidity were assessed retrospectively with regard to blastocyst, pregnancy and miscarriage rates. Its influence over morphokinetic parameters was evaluated by using the time-lapse system. Additionally, a preliminary study regarding the oxidative status of the spent embryo culture media was conducted using the TCL (Thermochemiluminescence) Analyzer TM.

Delayed blastulation is linked with changes at early cleavage stage detected by continuous embryo monitoring

Embryo development events Feature Total no. of events Detected by IVI Detected by bGA 2.0 Detected by both IVI-no annotation bGA 2.0-no annotation PN appear 311 300 (96%) 290 (93%) 279 (90%) 11 (4%) 21 (7%) PN disappear 311 288 (93%) 294 (95%) 272 (87%) 22 (7%) 16 (5%) 2-cell 311 304 (98%) 309 (99%) 302 (97%) 7 (2%) 2 (1%) 3-cell 311 302 (97%) 307 (99%) 298 (96%) 9 (3%) 4 (1%) 4-cell 311 299 (96%) 307 (99%) 297 (95%) 10 (3%) 2 (1%) 5-cell 311 295 (95%) 306 (98%) 293 (94%) 13 (4%) 2 (1%) 6-cell 311 288 (93%) 300 (96%) 281 (90%) 19 (6%) 7 (2%) Morula 311 218 (70%) 276 (89%) 214 (69%) 62 (20%) 4 (1%) Early B 311 188 (60%) 252 (81%) 177 (57%) 75 (24%) 11 (4%) All events 2799 2482 (89%) 2641 (94%) 2413 (86%) 228 (8%) 69 (2%) e218 ASRM Abstracts Vol. 110, No. 4, Supplement, September 2018