Optimizing Non-Invasive PGT-A: A Multi-Factorial Approach for Enhanced Accuracy and Seamless Integration Into Clinical IVF.

Purpose: This study aimed to identify factors influencing the accuracy and effectiveness of non-invasive preimplantation genetic testing for aneuploidy (niPGT-A) and develop clinical integration workflows to provide alternative embryo assessment and improve pregnancy outcomes. Methods: A two-phase study analyzed 341 blastocysts and spent culture medium (SCM) from 90 IVF patients. Phase 1 assessed niPGT-A accuracy by comparing SCM/trophectoderm (TE) with whole blastocysts (100 frozen embryos). Phase 2 examined concordance, assisted hatching, and culture-duration effects, focusing on outcomes after frozen embryo transfer using 241 fresh embryos. Nineteen euploid-TE/euploid-SCM and 14 euploid-TE/aneuploid-SCM blastocysts were transferred, and a meta-analysis was conducted using data from 163 euploid embryos across four studies. Results: The optimized niPGT-A workflow achieved a superior PPV of 92.1% and accuracy of 91.3%, outperforming conventional PGT-A. Ploidy and sex concordances were 76.5% and 98.9% among 183 fresh embryos. Assisted hatching improved amplification rates, while culture duration/ploidy did not impact concordance. Meta-analysis showed SCM-guided embryo transfer significantly improved pregnancy outcomes, compared to TE biopsy. Conclusion: This optimized niPGT-A procedure provides a highly accurate chromosomal assessment, seamlessly integrates with IVF workflows. It supports informed embryo transfer decisions, improves outcomes, reduces embryo damage risk, and limits invasive procedures, enhancing clinical management in reproductive medicine.