AUTHOR=Satirapod Chonthicha , Niransuk Pornsri , Tantanavipas Siriluk , Sensorn Insee , Chantratita Wasun , Trachoo Objoon 
  
TITLE=Application of non-invasive preimplantation genetic screening for aneuploidy through spent embryo culture media analysis at 48 and 54 hours after embryo cleavage
  
JOURNAL=British Journal of Biomedical Science
  
VOLUME=Volume 83 - 2026
  
YEAR=2026
  
URL=https://www.frontierspartnerships.org/journals/british-journal-of-biomedical-science/articles/10.3389/bjbs.2026.15905
  
DOI=10.3389/bjbs.2026.15905
  
ISSN=2474-0896
  
ABSTRACT=BackgroundThis study aims to assess the effectiveness of non-invasive preimplantation genetic testing (niPGT) for detecting aneuploidy at two different time points, 48- and 54-hour following the cleavage stage of embryo development.MethodsA cohort of 15 infertile women was enrolled, involving a total of 58 embryos. All embryos underwent sequential culture media procedures and received assisted hatching during the cleavage stage on Day 3. After biopsy examination, conducted either 48 or 54 hours post cleavage, spent culture media (SCM) were gathered and processed for the amplification and quantification of cell-free DNA. This was followed by low-pass whole genome sequencing.ResultsThe cell-free DNA content within SCM remained consistent across both time points post cleavage. The accuracy of niPGT in ploidy detection, in comparison to trophectoderm biopsy, was 53.71%. No significant distinction in ploidy detection accuracy was observed between SCM collected from embryos at 48 hours versus those at 54 hours post cleavage. The overall accuracy for sex determination reached 79.63%.ConclusionsThe concentration of cell-free DNA within SCM was found to be consistent at both 48- and 54-hours after embryo cleavage. However, the accuracy of ploidy determination, when contrasted with the conventional trophectoderm biopsy, did not yield satisfactory outcomes.