Author(s):
Lama Khudari, Marwan Halabi, Sahar Al Fahoum
Email(s):
lama.khudari@damascusuniversity.edu.sy
DOI:
10.52711/0974-360X.2026.00421 
Address:
Lama Khudari1*, Marwan Halabi2, Sahar Al Fahoum1
1Lama Khudari, Dept. of Biochemistry and Microbiology, Faculty of Pharmacy, Damascus University, Damascus, Syrian Arab Republic.
2Marwan Halabi, Dept. of Anatomy, Histology and Embryology, Faculty of Medicine, Damascus University, Damascus, Syrian Arab Republic.
3Sahar Al Fahoum, Dept. of Biochemistry and Microbiology, Faculty of Pharmacy, Damascus University, Damascus, Syrian Arab Republic.
*Corresponding Author
Published In:
Volume - 19,
Issue - 7,
Year - 2026
ABSTRACT:
Background: Preimplantation genetic screening (PGS) is essential in assisted reproductive technology (ART), enabling couples to assess embryos for genetic abnormalities before implantation during in vitro fertilization (IVF). This study investigates and validates a protocol for isolating cell-free fetal DNA (cffDNA) from spent blastocyst media, positioning it as a non-invasive biomarker for evaluating embryo chromosomal integrity. Our research aims to deepen the understanding of embryonic health and viability, which could lead to improved reproductive outcomes. By focusing on the use of cffDNA in spent culture media, this study addresses a significant gap in reproductive medicine, offering new insights into embryo assessment and potentially enhancing the selection process for viable embryos. Methods: This is a methodological cross-sectional study. We selected 21 women under 43 years old and collected 27 SBM samples from blastocyst-stage embryos. We tested various collection variables, including SCM volume, timing (Day 3 vs. Day 5), and storage conditions (fresh vs. frozen). We refined our cffDNA isolation technique and validated it via 24-chromosome aneuploidy screening using Next Generation Sequencing (NGS) on five SCM samples. Results: Our study successfully extracted free embryonic DNA (cffDNA) from media collected on day 5 of blastocyst culture, particularly utilizing fresh spent blastocyst medium (SBM) samples. We identified that the optimal volume for these SBM samples is less than 20 µl, with concentration values ranging from 2.59 to 56 ng/µl. Additionally, we found that the most effective method for isolating cffDNA from SBM is DOP-PCR, which is well-suited for amplifying the entire gDNA. Gel electrophoresis revealed amplification products as smears ranging from 200 bp to 2000 bp, indicating successful amplification. Regarding aneuploidy screening for the five SBM samples, our optimized protocol proved effective. We identified three euploid samples, categorized as normal female (XX), alongside one aneuploid sample (7:1,18:1, X0). One additional sample displayed a noisy NGS profile. Conclusion: Our results emphasize the reliability of our protocol for extracting cffDNA from spent Blastocyst Medium (SBM) and assessing the genetic status of the embryos, providing valuable insights for future research in reproductive genetics.
Cite this article:
Lama Khudari, Marwan Halabi, Sahar Al Fahoum. Optimization of Detecting Embryonic cell free DNA in Spent Blastocyst Media: A Non-invasive Approach for Preimplantation Genetic Screening. Research Journal of Pharmacy and Technology. 2026;19(7):2954-0. doi: 10.52711/0974-360X.2026.00421
Cite(Electronic):
Lama Khudari, Marwan Halabi, Sahar Al Fahoum. Optimization of Detecting Embryonic cell free DNA in Spent Blastocyst Media: A Non-invasive Approach for Preimplantation Genetic Screening. Research Journal of Pharmacy and Technology. 2026;19(7):2954-0. doi: 10.52711/0974-360X.2026.00421 Available on: https://rjptonline.org/AbstractView.aspx?PID=2026-19-7-8
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