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Updated: Sep 18, 2025
Chromosome Screening of Human Preimplantation Embryos by Using Spent Culture Medium: Sample Collection and Chromosomal Ploidy Analysis
Published on: September 7, 2021
Library concentration of cell-free DNA in spent culture medium: a potential indicator for clinical outcomes of
Jin Huang1, Yaxin Yao2, Jialin Jia1
1Centre for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China; National Clinical Research Center for Obstetrics and Gynecology, Peking University Third Hospital, Beijing, People's Republic of China.; State Key Laboratory of Female Fertility Promotion, Department of Obstetrics and Gynecology, Peking University Third Hospital, Peking, People's Republic of China; Key Laboratory of Assisted Reproduction (Peking University), Ministry of Education, Beijing, People's Republic of China; Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive, Technology, Beijing, People's Republic of China.
Research Question:
Can the library concentration of cell-free DNA (cfDNA) in spent culture medium (SCM) be a potential indicator of pregnancy outcomes after blastocyst transfer?
Design:
This retrospective observational study included 341 infertile couples undergoing intracytoplasmic sperm injection in which the female partner was over 35 years old. The blastocysts obtained from the enrolled participants were cultured individually and the associated SCM collected. Single-cell whole-genome amplification and DNA library preparation were performed, and the cfDNA concentration and the copy-number variation results for each sample analysed. A follow-up study was conducted after the blastocyst transfers. The association between library concentration and clinical outcome was analysed.
Results:
A total of 644 single vitrified-warmed blastocyst transfers were performed in 341 patients. The blastocysts were divided into low- and high-concentration groups according to the DNA library concentration, with a threshold of 1 ng/µl in the SCM. Considering all 644 embryo transfers, the rates of clinical pregnancy (43.65% versus 31.54%, P = 0.003), ongoing pregnancy (37.56% versus 23.49%, P < 0.001) and live birth (37.56% versus 23.04%, P < 0.001) were significantly higher in the low-concentration group versus the high-concentration group. For the 341 first-embryo transfers, rates of ongoing pregnancy (39.52% versus 28.57%, P = 0.038) and live birth (39.52% versus 28.11%, P = 0.030), but not clinical pregnancy (47.58% versus 37.33%, P = 0.064), were significantly higher.
Conclusions:
Library concentration may be a potentially important predictive indicator for blastocyst transfer. Given its shorter testing duration and potential to improve pregnancy outcomes, the use of library concentration to guide the transfer priority of embryos in fresh cycles could be considered.
