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The integrity of cfDNA in follicular fluid and spent medium from embryo culture is associated with embryo grade in patients undergoing in vitro fertilization

  • Embryo Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

This study was conducted to verify if the cfDNA integrity (cfDI) in follicular fluid and subsequent spent embryo medium (SEM) could serve as potential non-invasive biomarker for high-grade embryo selection during IVF/ICSI.

Methods

Thirty-two follicular fluids, 32 subsequent corresponding cleavage embryo SEM, and 23 subsequent blastocyst SEM were collected from 11 patients undergoing IVF/ICSI. CfDI was measured by ALU gene amplicons with different sizes by qPCR, as the ratio of long to short fragments.

Results

CfDI in follicular fluid corresponding to subsequent high-grade cleavage embryos and blastocysts was significantly lower than that related to low-grade embryos (p = 0.018). Conversely, cfDI in SEM was significantly and positively correlated with high-grade embryos at both stages (p = 0.009). ROC curves of the analysis of cfDI in follicular fluid showed great potential in predicting subsequent embryogenesis and embryo grade (AUC > 0.927). Regardless of the cleavage embryo grade by morphology, cfDI in day 3 SEM could predict if the cleavage embryo could develop to a high-grade blastocyst (AUC = 0.820). A concordant shift pattern of cfDI from follicular fluid to subsequent day 3 SEM and day 5 SEM was found in 81.82% participants featured by various clinical characteristics.

Conclusion

CfDI in follicular fluid and SEM was significantly correlated with embryogenesis and embryo grade and could serve as a potential non-invasive biomarker in high-grade embryo selection. Direct qPCR was proved as a labor-saving and sensitive method for the analysis of cfDI in low volume of SEM.

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Acknowledgements

We would like to thank the doctors in the Clinical Center of Reproductive Medicine, State Key Laboratory of Reproductive Medicine in the First Affiliated Hospital involved in the sample and data collection for their support in this study.

Funding

This study was supported by the National Natural Science Foundation of China (61801108) and the Natural Science Foundation of Jiangsu Province (BK20211166, BK20201148).

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Authors and Affiliations

  1. School of Medicine, Southeast University, Nanjing, China

    Min Pan

  2. State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China

    Huajuan Shi, Zhiyu Liu & Qinyu Ge

  3. Clinical Center of Reproductive Medicine, State Key Laboratory of Reproductive Medicine, First Affiliated Hospital, Nanjing Medical University, Nanjing, China

    Juan Dong & Lingbo Cai

Authors
  1. Min Pan
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  2. Huajuan Shi
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  4. Juan Dong
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Contributions

Min Pan and Qinyu Ge led the conception and design of the study. Lingbo Cai collected the follicular fluid and SEM samples and clinical data. Min Pan and Huajuan Shi carried out the evaluation of cfDI in samples. Zhiyu Liu carried out the direct qPCR experiments. Min Pan and Qinyu Ge conducted the data analysis, construction of tables and figures, and article revision. Lingbo Cai was involved in study conception and critical revision of the article. All authors approved the final article.

Corresponding authors

Correspondence to Lingbo Cai or Qinyu Ge.

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Ethics approval was obtained from the Clinical Center of Reproductive Medicine, First Affiliated Hospital, Nanjing in 2020.

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The authors declare competing interests.

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Pan, M., Shi, H., Liu, Z. et al. The integrity of cfDNA in follicular fluid and spent medium from embryo culture is associated with embryo grade in patients undergoing in vitro fertilization. J Assist Reprod Genet 38, 3113–3124 (2021). https://doi.org/10.1007/s10815-021-02357-0

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