Abstract Study question How does the outcomes of ICSI-AOA and child health differ from those reported in the literature? Summary answer ICSI-AOA clinical outcomes, including fertilization, embryo quality, pregnancy, live birth rates, and anomaly rates in children, align with those reported for ICSI alone. What is known already The use of AOA to artificially induce calcium rises by employing calcium ionophores has been shown as an effective approach in overcoming fertilization failure following ICSI, especially in couples facing severe male factor infertility. However, a significant gap exists in reporting live birth outcomes. On the other hand, uncertainties surrounding the health implications for children born through this method necessitate further investigation due to the limited sample sizes in previous reports. This descriptive analysis fills that gap by assessing 1300 cases, providing crucial insights into the clinical outcomes and health status of babies resulting from ICSI-AOA treatment. Study design, size, duration This retrospective study analyzes 1300 cases of ICSI-AOA between 2018 and 2020. Clinical outcomes in this study includes: fertilization, embryo quality, pregnancy, and live birth rates. Questionaries regarding the children health were included from birth to two years. Participants/materials, setting, methods The study included 1300 cases that referred to Isfahan Fertility and Infertility Center (Isfahan, Iran) and underwent ICSI-AOA due to: 1) cases with testicular biopsy (N = 458), 2) cases with 99-100% sperm head abnormal morphology and/or 3) previous low fertilization or total fertilization failure after ICSI (N = 842). In all cases, to induce AOA, oocytes were subjected to a 10-minute exposure of 10 micromolar ionomycin following ICSI. Main results and the role of chance From 1300 couples, 11,756 mature, 2,012 immatures, 193 degenerate, and 76 uninjectable oocytes were obtained. Fertilization were 59.69%, and 52.18% for mature and immature oocytes, resulting in cleavage rates of 96.93% and 93.80%, respectively. On day 3, 25.14% grade A, 31.49% grade B, and 43.36% grade C were observed. In 321 couples, 571 fresh embryos were transferred on day 3, resulting in clinical pregnancy and live birth rates of 20.82%, and 19.55%, respectively. For 32 couples with 44 fresh ET on day 5, rates were 9.38%, and 9.38% respectively. Out of 3270 vitrified embryos on day 3, 1743 were thawed (337 couples) and 1147 were transferred in 643 ET cycles. Clinical pregnancy and live birth rates were 21.61%, and 20.37%, respectively. 1527 embryos remain frozen. Out of 2,974 day 5-embryos, 2,571 did not reach blastocyst, leaving 403 vitrified embryos in 699 cases. Among them, 197 embryos remain frozen (80 cases) while 206 were transferred, with clinical pregnancy and live birth rates of 22.11%, and 21.80%, respectively. Cancellation rate, encompassing failed fertilization, cancellations on multiple days, and after freeze-thawing, was 20.16%. Among 266 live births, seven children showed anomalies, including achondroplasia, combined immunodeficiency, left eye deviation, intrauterine growth restriction, hydrocephaly, epilepsy, and umbilical hernia. Limitations, reasons for caution The main limitations of this study were the lack of comprehensive information on previous cycles, the absence of an ICSI control group, and the limited long-term follow-up on the physical and mental health of children born through the ICSI-AOA method beyond age two. Wider implications of the findings ICSI-AOA yields helpful clinical outcomes in high fertilization, pregnancy, and live birth rates in infertile couples, especially those with severe male factor infertility similar to ICSI in literature. Furthermore, presently, there seem to be no real concerns about health of babies born through this method, aligning with ICSI studies. Trial registration number IR.ACECR.ROYAN.REC.1401.038
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