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CC BY 4.0 · Glob Med Genet 2024; 11(03): 227-232
DOI: 10.1055/s-0044-1788260
Original Article

Analyzing Cell-free Genomic DNA in Spent Culture Media: Noninvasive Insight into the Blastocysts

Siddhartha Shankar Layek
1   National Dairy Development Board, Anand, Gujarat, India
,
Shrushti Kanani
2   Department of Clinical Embryology, MGM School of Biomedical Sciences, MGM Institute of Health Sciences, Kamothe, Navi Mumbai, Maharashtra, India
,
Shilpa Doultani
3   Department of Zoology, Gujarat University, Navrangpura, Ahmedabad, Gujarat, India
,
Tejas Gohil
4   Sabarmati Ashram Gaushala, Kheda, Gujarat, India
,
Sanket Patil
1   National Dairy Development Board, Anand, Gujarat, India
,
Ananthasayanam Sudhakar
1   National Dairy Development Board, Anand, Gujarat, India
,
Kathan Banubhai Raval
1   National Dairy Development Board, Anand, Gujarat, India
,
Karuppanasamy Kuppusamy
1   National Dairy Development Board, Anand, Gujarat, India
,
Sanjay Gorani
1   National Dairy Development Board, Anand, Gujarat, India
,
Sudharson Raj
4   Sabarmati Ashram Gaushala, Kheda, Gujarat, India
,
Rafiya Sangameshwari
2   Department of Clinical Embryology, MGM School of Biomedical Sciences, MGM Institute of Health Sciences, Kamothe, Navi Mumbai, Maharashtra, India
,
Himali Jadeja
2   Department of Clinical Embryology, MGM School of Biomedical Sciences, MGM Institute of Health Sciences, Kamothe, Navi Mumbai, Maharashtra, India
,
Mini Mol P.
5   Department of Anatomy, MGM Medical College, MGM Institute of Health Sciences, Kamothe, Navi Mumbai, Maharashtra, India
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Abstract

A commonly accepted standard protocol for noninvasive techniques for the genetic evaluation of an embryo remains elusive due to inconclusiveness regarding the volume of spent media to be acquired and the possibility of acquiring the same for subsequent analysis. Single embryo culture is imperative for standardizing noninvasive preimplantation testing using cell-free DNA (cf-DNA) released by individual developing embryos. This study aims to compare the development dynamics of single-drop embryonic culture against with group embryonic culture to establish a standardized protocol for noninvasive Preimplantation Genetic Testing (PGT) in bovine. A total of 239 cumulus–oocyte complexes were aspirated and subjected to in vitro maturation and fertilization. Among these, 120 embryos of day 3 were transferred to single-drop culture until the blastocyst stage. The single-drop culture drops were prepared using microdrops of 30 μL. At the blastocyst stage, spent media from all single-drop embryos were utilized for extracting cell-free genomic DNA to standardize the protocol. The blastocyst rate indicates no significant difference between the two culture methods, suggesting that single-drop culture is suitable for the process. Additionally, the extracted spent media yielded sufficient quantities of cf-DNA, supporting its potential use for PGT (p < 0.05). These findings support the hypothesis that single-drop embryo culture is a viable method for cf-DNA extraction and confirm the potential of using DNA fragments from spent media as a reliable source for noninvasive PGT.

Keywords

single embryo culture - cf-DNA - spent media - noninvasive PGT - blastocysts

Ethical Approval

Ethics approval was granted by the MGM Institutional Ethical Committee.




Publication History

Article published online:
23 July 2024

© 2024. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

 

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