Cell-free DNA in Embryo Culture Media as Non-invasive Biomarker of the Quality of Embryo Cleavage
Annual Research & Review in Biology,
Page 12-18
DOI:
10.9734/arrb/2023/v38i230569
Abstract
Background: The success of in vitro-fertilization (IVF) cycles is determined in large part by the quality of embryo cleavage, which in turn, is dependent on the quality of the embryo culture media (CM). Many factors can influence the quality of embryo CM, one of which is the levels of Cell Free Deoxyribonucleic acid (DNA). Understanding the association between Cell-free DNA levels in embryo CM and the quality of embryo cleavage could help improve the quality of IVF techniques.
Methods: This prospective study was conducted with 96 spent CM from patients undergoing IVF cycle, in order to determine relationships of Cell-free DNA levels in embryo CM with embryo cleavage quality on day 3. After intracytoplasmic sperm injection (ICSI), 48 embryos were evaluated on day 3 of their development, according to their cell number. Day 2 and day 3 CM corresponding to each one of the embryos was analyzed, by quantitative PCR, for estimation of Cell-free DNA levels.
Results: The results revealed a significant increase in Cell-free DNA levels on day 2 CM corresponding to 4 to 6 cell embryos compared to those corresponding to 7 to 8 cell embryos (p=0.04). As for day 3 CM, the results showed no significant difference between the Cell-Free DNA levels in CM of 7-8 and those of 4-6 cell embryos (p=0.4). Also, cell free DNA levels in embryo CM, were significantly higher on day 2 compared to day 3 for both 7-8 and 4-6 cell embryos (p=0.03; p=0.04).
Conclusion: We conclude that cell-free DNA levels in CM might be associated with delayed embryo cleavage.
Keywords:
- Cell-free DNA
- embryo cleavage
- embryo culture
- in vitro fertilization
How to Cite
Jamil, M., Debbarh, H., Jelloul, H., Kabit, A., Ennaji, M., Zarqaoui, M., Hissane, E. M., Senhaji, W., Louanjli, N., & Cadi, R. (2023). Cell-free DNA in Embryo Culture Media as Non-invasive Biomarker of the Quality of Embryo Cleavage. Annual Research & Review in Biology, 38(2), 12-18. https://doi.org/10.9734/arrb/2023/v38i230569
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Yu CH, Zhang RP, Li J. A predictive model for high-quality blastocyst based on blastomere number, fragmentation, and symmetry. Journal of Assisted Reproduction and Genetics. 2018;35:809-816.
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Milewski R, Ajduk A. Time-lapse imaging of cleavage divisions in embryo quality assessment. Reproduction. 2017;154(2):R37-R53.
Lundin K, Bergh C, Hardarson T. Early embryo cleavage is a strong indicator of embryo quality in human IVF. Human Reproduction. 2001;16(12):2652-2657.
Shamonki MI, Jin H, Haimowitz Z, Liu L. Proof of concept: preimplantation genetic screening without embryo biopsy through analysis of cell-free DNA in spent embryo culture media. Fertility and Sterility. 2016;106(6):1312-1318.
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Rubio C, et al. Embryonic cell-free DNA versus trophectoderm biopsy for aneuploidy testing: Concordance rate and clinical implications. Fertility and Sterility. 2019;112(3):510-519.
Scalici E, Traver S, Molinari N, Mullet T, Monforte M, Vintejoux E, Hamamah S. Cell-free DNA in human follicular fluid as a biomarker of embryo quality. Human Reproduction. 2014;29(12) :2661-2669.
Kustanovich A, Schwartz R, Peretz T, Grinshpun A. Life and death of circulating cell-free DNA. Cancer Biology & Therapy. 2019;20(8):1057-1067.
Bronkhorst AJ, Aucamp J, Pretorius PJ. Cell-free DNA: preanalytical variables. Clinica Chimica Acta. 2015;450:243-253.
Johnson KC, Verhaak RG. Serum cell-free DNA epigenetic biomarkers aid glioma diagnostics and monitoring. Neuro-oncology. 2021;23(9):1423-1424.
Duvvuri B, Lood C. Cell-free DNA as a biomarker in autoimmune rheumatic diseases. Frontiers in Immunology. 2019; 10:502.
Johnston DG, Hambly R, Kearney N, Tobin DJ, Kirby B. Cell-free DNA is elevated in the serum of patients with hidradenitis suppurativa. The Journal of Dermatology; 2022.
Szilágyi M, et al. Circulating cell-free nucleic acids: Main characteristics and clinical application. International Journal of Molecular Sciences. 2020;21(18):6827.
Hu Z, Chen H, Long Y, Li P, Gu Y. The main sources of circulating cell-free DNA: Apoptosis, necrosis and active secretion. Critical Reviews in Oncology/ Hematology. 2021;157:103166.
Tsuji N, Agbor-Enoh S. Cell-free DNA beyond a biomarker for rejection: Biological trigger of tissue injury and potential therapeutics. The Journal of Heart and Lung Transplantation. 2021; 40(6):405-413.
Hammond ER, Shelling AN, Cree LM. Nuclear and mitochondrial DNA in blastocoele fluid and embryo culture medium: Evidence and potential clinical use. Human Reproduction. 2016;31(8): 1653-1661.
Voet T, Vanneste E, Vermeesch JR. The human cleavage stage embryo is a cradle of chromosomal rearrangements. Cytogenetic and Genome Research. 2011;133(2-4):160-168.
Fadeel B, Orrenius S. Apoptosis: a basic biological phenomenon with wide‐ranging implications in human disease. Journal of Internal Medicine. 2005;258(6):479-517.
Heitzer E, Auinger L, Speicher MR. Cell-free DNA and apoptosis: How dead cells inform about the living. Trends in Molecular Medicine. 2020;26(5):519-528.
Ramos-Ibeas P, Gimeno I, Cañón-Beltrán K, Gutiérrez-Adán A, Rizos D, Gómez E. Senescence and apoptosis during in vitro embryo development in a bovine model. Frontiers in Cell and Developmental Biology. 2020;8:619902.
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Rostami A, Lambie M, Caberry WY, Stambolic V, Waldron JN, Bratman SV. Senescence, necrosis, and apoptosis govern circulating cell-free DNA release kinetics. Cell Reports. 2020;31(13): 107830.
Shitara A, et al. Cell-free DNA in spent culture medium effectively reflects the chromosomal status of embryos following culturing beyond implantation compared to trophectoderm biopsy. Plos One. 2021;16 (2):e0246438.
Kong X, Yang S, Gong F, Lu C, Zhang S, Lu G, Lin G. The relationship between cell number, division behavior and developmental potential of cleavage stage human embryos: A time-lapse study. Plos One. 2016;11(4):e0153697.
Magli MC, Gianaroli L, Ferraretti AP, Lappi M, Ruberti A, Farfalli V. Embryo morphology and development are dependent on the chromosomal complement. Fertility and Sterility. 2007;87 (3):534-541.
Jaroudi S, SenGupta S. DNA repair in mammalian embryos. Mutation Research/ Reviews in Mutation Research. 2007; 635(1):53-77.
Amir H, et al. Time-lapse imaging reveals delayed development of embryos carrying unbalanced chromosomal translocations. Journal of Assisted Reproduction and Genetics. 2019;36:315-324.
Jamil M, et al. Lipid Peroxidation Status in Embryo Culture Media: The Impact on Fertilization and Embryo Quality during IVF Cycles. Annual Research & Review in Biology, 2022 37(12), 61-74
Rahman T, Hosen I, Islam MT, Shekhar HU. Oxidative stress and human health. Advances in Bioscience and Biotechnology. 2012;3:997-1019.
Hajam YA, et al. Oxidative stress in human pathology and aging: Molecular mechanisms and perspectives. Cells. 2022;11(3):552.
Amir H, et al. Time-lapse imaging reveals delayed development of embryos carrying unbalanced chromosomal translocations. Journal of Assisted Reproduction and Genetics. 2019;36:315-324
Reed ML, Hamic A, Thompson DJ, Caperton CL. Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: A sibling embryo study. Fertility and Sterility. 2009;92(5):1783-1786.
Mantikou E, Youssef MA, van Wely M, van der Veen F, Al-Inany HG, Repping S, Mastenbroek S. Embryo culture media and IVF/ICSI success rates: A systematic review. Human Reproduction Update. 2013;19(3):210-220.
Sunde A, et al. Time to take human embryo culture seriously. Human Reproduction. 2016;31(10):2174-2182.
Machtinger R, Racowsky C. Culture systems: Single step. Embryo Culture: Methods and Protocols. 2012;199-209.
Yu CH, Zhang RP, Li J. A predictive model for high-quality blastocyst based on blastomere number, fragmentation, and symmetry. Journal of Assisted Reproduction and Genetics. 2018;35:809-816.
Desai NN, Goldstein J, Rowland DY, Goldfarb JM. Morphological evaluation of human embryos and derivation of an embryo quality scoring system specific for day 3 embryos: A preliminary study. Human Reproduction. 2000;15(10): 2190-2196.
Prados FJ, Debrock S, Lemmen JG, Agerholm I. The cleavage stage embryo. Human Reproduction. 2012;27(suppl_1): i50-i71.
Milewski R, Ajduk A. Time-lapse imaging of cleavage divisions in embryo quality assessment. Reproduction. 2017;154(2):R37-R53.
Lundin K, Bergh C, Hardarson T. Early embryo cleavage is a strong indicator of embryo quality in human IVF. Human Reproduction. 2001;16(12):2652-2657.
Shamonki MI, Jin H, Haimowitz Z, Liu L. Proof of concept: preimplantation genetic screening without embryo biopsy through analysis of cell-free DNA in spent embryo culture media. Fertility and Sterility. 2016;106(6):1312-1318.
Brouillet S, Martinez G, Coutton C, Hamamah S. Is cell-free DNA in spent embryo culture medium an alternative to embryo biopsy for preimplantation genetic testing? A systematic review. Reproductive Biomedicine Online. 2020;40(6):779-796.
Rubio C, et al. Embryonic cell-free DNA versus trophectoderm biopsy for aneuploidy testing: Concordance rate and clinical implications. Fertility and Sterility. 2019;112(3):510-519.
Scalici E, Traver S, Molinari N, Mullet T, Monforte M, Vintejoux E, Hamamah S. Cell-free DNA in human follicular fluid as a biomarker of embryo quality. Human Reproduction. 2014;29(12) :2661-2669.
Kustanovich A, Schwartz R, Peretz T, Grinshpun A. Life and death of circulating cell-free DNA. Cancer Biology & Therapy. 2019;20(8):1057-1067.
Bronkhorst AJ, Aucamp J, Pretorius PJ. Cell-free DNA: preanalytical variables. Clinica Chimica Acta. 2015;450:243-253.
Johnson KC, Verhaak RG. Serum cell-free DNA epigenetic biomarkers aid glioma diagnostics and monitoring. Neuro-oncology. 2021;23(9):1423-1424.
Duvvuri B, Lood C. Cell-free DNA as a biomarker in autoimmune rheumatic diseases. Frontiers in Immunology. 2019; 10:502.
Johnston DG, Hambly R, Kearney N, Tobin DJ, Kirby B. Cell-free DNA is elevated in the serum of patients with hidradenitis suppurativa. The Journal of Dermatology; 2022.
Szilágyi M, et al. Circulating cell-free nucleic acids: Main characteristics and clinical application. International Journal of Molecular Sciences. 2020;21(18):6827.
Hu Z, Chen H, Long Y, Li P, Gu Y. The main sources of circulating cell-free DNA: Apoptosis, necrosis and active secretion. Critical Reviews in Oncology/ Hematology. 2021;157:103166.
Tsuji N, Agbor-Enoh S. Cell-free DNA beyond a biomarker for rejection: Biological trigger of tissue injury and potential therapeutics. The Journal of Heart and Lung Transplantation. 2021; 40(6):405-413.
Hammond ER, Shelling AN, Cree LM. Nuclear and mitochondrial DNA in blastocoele fluid and embryo culture medium: Evidence and potential clinical use. Human Reproduction. 2016;31(8): 1653-1661.
Voet T, Vanneste E, Vermeesch JR. The human cleavage stage embryo is a cradle of chromosomal rearrangements. Cytogenetic and Genome Research. 2011;133(2-4):160-168.
Fadeel B, Orrenius S. Apoptosis: a basic biological phenomenon with wide‐ranging implications in human disease. Journal of Internal Medicine. 2005;258(6):479-517.
Heitzer E, Auinger L, Speicher MR. Cell-free DNA and apoptosis: How dead cells inform about the living. Trends in Molecular Medicine. 2020;26(5):519-528.
Ramos-Ibeas P, Gimeno I, Cañón-Beltrán K, Gutiérrez-Adán A, Rizos D, Gómez E. Senescence and apoptosis during in vitro embryo development in a bovine model. Frontiers in Cell and Developmental Biology. 2020;8:619902.
Tjoa ML, Cindrova-Davies T, Spasic-Boskovic O, Bianchi DW, Burton GJ. Trophoblastic oxidative stress and the release of cell-free feto-placental DNA. The American Journal of Pathology. 2006;169(2):400-4.
Rostami A, Lambie M, Caberry WY, Stambolic V, Waldron JN, Bratman SV. Senescence, necrosis, and apoptosis govern circulating cell-free DNA release kinetics. Cell Reports. 2020;31(13): 107830.
Shitara A, et al. Cell-free DNA in spent culture medium effectively reflects the chromosomal status of embryos following culturing beyond implantation compared to trophectoderm biopsy. Plos One. 2021;16 (2):e0246438.
Kong X, Yang S, Gong F, Lu C, Zhang S, Lu G, Lin G. The relationship between cell number, division behavior and developmental potential of cleavage stage human embryos: A time-lapse study. Plos One. 2016;11(4):e0153697.
Magli MC, Gianaroli L, Ferraretti AP, Lappi M, Ruberti A, Farfalli V. Embryo morphology and development are dependent on the chromosomal complement. Fertility and Sterility. 2007;87 (3):534-541.
Jaroudi S, SenGupta S. DNA repair in mammalian embryos. Mutation Research/ Reviews in Mutation Research. 2007; 635(1):53-77.
Amir H, et al. Time-lapse imaging reveals delayed development of embryos carrying unbalanced chromosomal translocations. Journal of Assisted Reproduction and Genetics. 2019;36:315-324.
Jamil M, et al. Lipid Peroxidation Status in Embryo Culture Media: The Impact on Fertilization and Embryo Quality during IVF Cycles. Annual Research & Review in Biology, 2022 37(12), 61-74
Rahman T, Hosen I, Islam MT, Shekhar HU. Oxidative stress and human health. Advances in Bioscience and Biotechnology. 2012;3:997-1019.
Hajam YA, et al. Oxidative stress in human pathology and aging: Molecular mechanisms and perspectives. Cells. 2022;11(3):552.
Amir H, et al. Time-lapse imaging reveals delayed development of embryos carrying unbalanced chromosomal translocations. Journal of Assisted Reproduction and Genetics. 2019;36:315-324
Reed ML, Hamic A, Thompson DJ, Caperton CL. Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: A sibling embryo study. Fertility and Sterility. 2009;92(5):1783-1786.
Mantikou E, Youssef MA, van Wely M, van der Veen F, Al-Inany HG, Repping S, Mastenbroek S. Embryo culture media and IVF/ICSI success rates: A systematic review. Human Reproduction Update. 2013;19(3):210-220.
Sunde A, et al. Time to take human embryo culture seriously. Human Reproduction. 2016;31(10):2174-2182.
Machtinger R, Racowsky C. Culture systems: Single step. Embryo Culture: Methods and Protocols. 2012;199-209.
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