A. Tejera

The use of morphokinetics as a predictor of embryo implantation

BACKGROUND Time-lapse observation presents an opportunity for optimizing embryo selection based on morphological grading as well as providing novel kinetic parameters, which may further improve accurate selection of viable embryos. The objective of this retrospective study was to identify the morphokinetic parameters specific to embryos that were capable of implanting. In order to compare a large number of embryos, with minimal variation in culture conditions, we have used an automatic embryo monitoring system. METHODS Using a tri-gas IVF incubator with a built-in camera designed to automatically acquire images at defined time points, we have simultaneously monitored up to 72 individual embryos without removing the embryos from the controlled environment. Images were acquired every 15 min in five different focal planes for at least 64 h for each embryo. We have monitored the development of transferred embryos from 285 couples undergoing their first ICSI cycle. The total number of transferred embryos was 522, of which 247 either failed to implant or fully implanted, with full implantation meaning that all transferred embryos in a treatment implanted. RESULTS A detailed retrospective analysis of cleavage times, blastomere size and multinucleation was made for the 247 transferred embryos with either failed or full implantation. We found that several parameters were significantly correlated with subsequent implantation (e.g. time of first and subsequent cleavages as well as the time between cleavages). The most predictive parameters were: (i) time of division to 5 cells, t5 (48.8-56.6 h after ICSI); (ii) time between division to 3 cells and subsequent division to 4 cells, s2 (≤ 0.76 h) and (iii) duration of cell cycle two, i.e. time between division to 2 cells and division to 3 cells, cc2 (≤ 11.9 h). We also observed aberrant behavior such as multinucleation at the 4 cell stage, uneven blastomere size at the 2 cell stage and abrupt cell division to three or more cells, which appeared to largely preclude implantation. CONCLUSIONS The image acquisition and time-lapse analysis system makes it possible to determine exact timing of embryo cleavages in a clinical setting. We propose a multivariable model based on our findings to classify embryos according to their probability of implantation. The efficacy of this classification will be evaluated in a prospective randomized study that ultimately will determine if implantation rates can be improved by time-lapse analysis.

Successful pregnancy and childbirth after intracytoplasmic sperm injection with calcium ionophore oocyte activation in a globozoospermic patient

OBJECTIVE To check the effectiveness of intracytoplasmic sperm injection (ICSI) combined with assisted oocyte activation (AOA) in a globozoospermic patient. DESIGN Case report. SETTING Instituto Valenciano de Infertilidad, Valencia, Spain. PATIENT(S) A patient with globozoospermia. INTERVENTION(S) ICSI was administered in 14 oocytes. ICSI combined with AOA, in which a small amount of calcium was injected followed by calcium ionophore exposure, was done in 9 oocytes. MAIN OUTCOME MEASURE(S) Fertilization rate and embryo quality was assessed in both groups. RESULT(S) Chemical activation increased fertilization rate (55.6% vs. 35.7%) and the number of embryos with less multinucleation on day 2 (0 vs. 60%). Two embryos generated from AOA were transferred into the uterus (on day 3), resulting in a pregnancy and a healthy newborn. CONCLUSION(S) The AOA with calcium ionophore treatment improved fertilization rate and quality of the embryos, and was found to be an effective method for AOA in this patient with a low fertilization rate after previous ICSI treatment.

Time-dependent O2 consumption patterns determined optimal time ranges for selecting viable human embryos

OBJECTIVE To evaluate correlations between metabolic activity and implantation potential of transferred embryos in a study based on oxygen (O(2)) consumption (OC) measurements, because O(2) uptake is directly related to the capacity of an embryo to produce energy via adenosine triphosphate. DESIGN Retrospective cohort study. SETTING Infertility institute. PATIENT(S) Five hundred seventy-five injected oocytes in 56 first oocyte donation cycles with embryo transfer on day 3. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) We analyzed embryo destination viability and implantation depending on the embryo OC rate obtained from 47,741 measurements (up to 85 measurements per embryo, 2-3 measurements per hour). OC patterns were analyzed in relation to the time elapsed from sperm microinjection, to the final destination of the embryos (transferred, frozen, or discarded), to ongoing pregnancy, and by successful implantation. RESULT(S) OC was found to decrease during embryonic development. OC patterns from 52 hours onward showed the strongest correlation with implantation success. Regarding embryo destination, the same patterns were observed. CONCLUSION(S) OC from individual embryos revealed significant differences, mainly close to the time of transfer, when OC pattern was associated with successful implantation. Therefore, measuring the OC pattern of human embryos culture up to 72 hours could be used to select the embryo with best developmental potential.

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.