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Insights gained from a cleavage-stage embryo freeze-all policy in in vitro maturation cycles

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

Abstract

Purpose

Cycles of in vitro maturation (IVM) of oocytes show asynchrony between embryo development and endometrial receptivity. Hence, elective embryo vitrification/warming (V/W) for embryo transfer (FET) is routinely performed. However, clinical outcomes after IVM are lower compared to conventional ovarian stimulation, mainly due to lower embryo quality. Vitrification at cleavage stage, rather than blastocyst stage, is used to optimize embryo utilization while maintaining acceptable pregnancy rates. The aim of this study is to ascertain the vulnerability to V/W of IVM-derived cleavage-stage embryos and to identify characteristics that predict pregnancy.

Methods

In this single-center retrospective cohort study, 442 day-3 IVM-derived embryos from PCOS patients were investigated. Cell survival upon warming, cell cycle progression during overnight culture and clinical outcome in 425 FET cycles were analyzed.

Results

From 442 V/W embryos, 85% were fully intact. Cell loss reduced the cell cycle progression after overnight culture of V/W embryos (p = 0.047) and tended to lower clinical pregnancy rates (16% vs 23%, p = 0.22) compared to intact embryos. Better fresh embryo quality was associated with enhanced cell cycle resumption after overnight culture (p < 0.0001). Cell cycle resumption was required for pregnancy to occur. Additionally, the extent of cell cycle progression (OR = 0.439, CI = 0.24–0.78, p < 0.001) and the number of available top-quality embryos (OR = 0.174, CI = 0.04–0.32, p = 0.01) were indicative for success.

Conclusions

A clinical pregnancy rate of 23% per FET with V/W IVM-derived cleavage-stage embryo was obtained with cell cycle resumption after overnight culture as dominant predictive factor.

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Data availability

Data are available upon request.

References

  1. Mostinckx L, Goyens E, Mackens S, Roelens C, Boudry L, Uvin V, et al. Clinical outcomes from ART in predicted hyperresponders: in vitro maturation of oocytes versus conventional ovarian stimulation for IVF/ICSI. Hum Reprod. 2024;39:586–94.

    CAS  PubMed  Google Scholar 

  2. Galvão A, Segers I, Smitz J, Tournaye H, De Vos M. In vitro maturation (IVM) of oocytes in patients with resistant ovary syndrome and in patients with repeated deficient oocyte maturation. J Assist Reprod Genet. 2018;35:2161–71.

    PubMed  PubMed Central  Google Scholar 

  3. Segers I, Bardhi E, Mateizel I, Van Moer E, Schots R, Verheyen G, et al. Live births following fertility preservation using in-vitro maturation of ovarian tissue oocytes. Hum Reprod. 2020;35:2026–36.

    PubMed  Google Scholar 

  4. De Vos M, Grynberg M, Ho TM, Yuan Y, Albertini DF, Gilchrist RB. Perspectives on the development and future of oocyte IVM in clinical practice. J Assist Reprod Genet. 2021;38:1265–80.

    PubMed  PubMed Central  Google Scholar 

  5. Braam SC, Ho VNA, Pham TD, Mol BW, van Wely M, Vuong LN. In-vitro maturation versus IVF: a cost-effectiveness analysis. Reprod Biomed Online. 2021;42:143–9.

    PubMed  Google Scholar 

  6. Gilchrist RB, Smitz J. Oocyte in vitro maturation: physiological basis and application to clinical practice. Fertil Steril. 2023;119:524–39.

    PubMed  Google Scholar 

  7. Vuong LN, Pham TD, Ho TM, De Vos M. Outcomes of clinical in vitro maturation programs for treating infertility in hyper responders: a systematic review. Fertil Steril. 2023;119:540–9.

    CAS  PubMed  Google Scholar 

  8. Pham HH, Le AH, Nguyen AM, Ha UD, Nguyen TC, Pham TD, et al. Cumulative live birth rate after oocyte in vitro maturation with a pre-maturation step in women with polycystic ovary syndrome or high antral follicle count. J Assist Reprod Genet. 2023;40:827–35.

    PubMed  PubMed Central  Google Scholar 

  9. Piechota S, Marchante M, Giovannini A, Paulsen B, Potts KS, Rockwell G, et al. Human-induced pluripotent stem cell-derived ovarian support cell co-culture improves oocyte maturation in vitro after abbreviated gonadotropin stimulation. Hum Reprod. 2023;38:2456–69.

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Ho VNA, Braam SC, Pham TD, Mol BW, Vuong LN. The effectiveness and safety of in vitro maturation of oocytes versus in vitro fertilization in women with a high antral follicle count. Hum Reprod. 2019;34:1055–64.

    PubMed  Google Scholar 

  11. Zheng X, Guo W, Zeng L, Zheng D, Yang S, Xu Y, et al. In vitro maturation without gonadotropins versus in vitro fertilization with hyperstimulation in women with polycystic ovary syndrome: a non-inferiority randomized controlled trial. Hum Reprod. 2022;37:242–53.

    CAS  PubMed  Google Scholar 

  12. Glujovsky D, QuinteiroRetamar AM, Alvarez Sedo CR, Ciapponi A, Cornelisse S, Blake D. Cleavage-stage versus blastocyst-stage embryo transfer in assisted reproductive technology. Cochrane Database Syst Rev. 2022;5:CD002118.

    PubMed  Google Scholar 

  13. De Vos A, Van Landuyt L, Santos-Ribeiro S, Camus M, Van de Velde H, Tournaye H, et al. Cumulative live birth rates after fresh and vitrified cleavage-stage versus blastocyst-stage embryo transfer in the first treatment cycle. Hum Reprod. 2016;31:2442–9.

    PubMed  Google Scholar 

  14. De Croo I, Colman R, De Sutter P, Stoop D, Tilleman K. No difference in cumulative live birth rates between cleavage versus blastocyst transfer in patients with four or fewer zygotes: results from a retrospective study. Hum Reprod Open. 2022;2022:hoac031.

    PubMed  PubMed Central  Google Scholar 

  15. Ortega-Hrepich C, Stoop D, Guzmán L, Van Landuyt L, Tournaye H, Smitz J, et al. A “freeze-all” embryo strategy after in vitro maturation: a novel approach in women with polycystic ovary syndrome? Fertil Steril. 2013;100:1002–7.

    PubMed  Google Scholar 

  16. Ortega-Hrepich C, Drakopoulos P, Bourgain C, Van Vaerenbergh I, Guzman L, Tournaye H, et al. Aberrant endometrial steroid receptor expression in in-vitro maturation cycles despite hormonal luteal support: A pilot study. Reprod Biol. 2019;19:210–7.

    PubMed  Google Scholar 

  17. Van Landuyt L, Van de Velde H, De Vos A, Haentjens P, Blockeel C, Tournaye H, et al. Influence of cell loss after vitrification or slow-freezing on further in vitro development and implantation of human Day 3 embryos. Hum Reprod. 2013;28:2943–9.

    PubMed  Google Scholar 

  18. Maheshwari A, Pandey S, Amalraj Raja E, Shetty A, Hamilton M, Bhattacharya S. Is frozen embryo transfer better for mothers and babies? Can cumulative meta-analysis provide a definitive answer? Hum Reprod Update. 2018;24:35–58.

    PubMed  Google Scholar 

  19. Pinborg A, Løssl K. Efficiency and drawbacks of frozen embryo transfers from artificial versus natural cycles. Lancet. 2024;404:219–20.

    PubMed  Google Scholar 

  20. Di Guardo F, Racca A, Coticchio G, Borini A, Drakopoulos P, Mackens S, et al. Impact of cell loss after warming of human vitrified day 3 embryos on obstetric outcome in single frozen embryo transfers. J Assist Reprod Genet. 2022;39:2069–75.

    PubMed  PubMed Central  Google Scholar 

  21. Aihaiti R, Shen Z, Wu X, Niu Z. Pregnancy complications and birth outcomes in women with polycystic ovary syndrome undergoing frozen embryo transfer. Fertil Steril. 2024;S0015–0282(24):00613–7.

    Google Scholar 

  22. Teede HJ, Tay CT, Laven J, Dokras A, Moran LJ, Piltonen TT, et al. Recommendations from the 2023 international evidence-based guideline for the assessment and management of polycystic ovary syndrome. Fertil Steril. 2023;120:767–93.

    PubMed  Google Scholar 

  23. Mackens S, Mostinckx L, Drakopoulos P, Segers I, Santos-Ribeiro S, Popovic-Todorovic B, et al. Early pregnancy loss in patients with polycystic ovary syndrome after IVM versus standard ovarian stimulation for IVF/ICSI. Hum Reprod. 2020;35:2763–73.

    CAS  PubMed  Google Scholar 

  24. Van Landuyt L, Polyzos NP, De Munck N, Blockeel C, Van de Velde H, Verheyen G. A prospective randomized controlled trial investigating the effect of artificial shrinkage (collapse) on the implantation potential of vitrified blastocysts. Hum Reprod. 2015;30:2509–18.

    PubMed  Google Scholar 

  25. Jiang S, Jin W, Zhao X, Xi Q, Chen L, Gao Y, et al. The impact of blastomere loss on pregnancy and neonatal outcomes of vitrified-warmed Day3 embryos in single embryo transfer cycles. J Ovarian Res. 2022;15:62.

    CAS  PubMed  PubMed Central  Google Scholar 

  26. De Vos A, Staessen C, De Rycke M, Verpoest W, Haentjens P, Devroey P, et al. Impact of cleavage-stage embryo biopsy in view of PGD on human blastocyst implantation: a prospective cohort of single embryo transfers. Hum Reprod. 2009;24:2988–96.

    PubMed  Google Scholar 

  27. Wissing ML, Bjerge MR, Olesen AIG, Hoest T, Mikkelsen AL. Impact of PCOS on early embryo cleavage kinetics. Reprod Biomed Online. 2014;28:508–14.

    CAS  PubMed  Google Scholar 

  28. Roesner S, Dietrich JE, Weigert J, Montag M, Toth B, Strowitzki T. Time-lapse imaging reveals differences in growth dynamics of embryos after in vitro maturation compared with conventional stimulation. Fertil Steril. 2017;107:606-612.e3.

    PubMed  Google Scholar 

  29. Walls ML, Hart R, Keelan JA, Ryan JP. Structural and morphologic differences in human oocytes after in vitro maturation compared with standard in vitro fertilization. Fertil Steril. 2016;106:1392-1398.e5.

    PubMed  Google Scholar 

  30. Edgar DH, Bourne H, Jericho H, McBain JC. The developmental potential of cryopreserved human embryos. Mol Cell Endocrinol. 2000;169:69–72.

    CAS  PubMed  Google Scholar 

  31. Nagy ZP, Taylor T, Elliott T, Massey JB, Kort HI, Shapiro DB. Removal of lysed blastomeres from frozen-thawed embryos improves implantation and pregnancy rates in frozen embryo transfer cycles. Fertil Steril. 2005;84:1606–12.

    PubMed  Google Scholar 

  32. Tšuiko O, Catteeuw M, Zamani Esteki M, Destouni A, Bogado Pascottini O, Besenfelder U, et al. Genome stability of bovine in vivo-conceived cleavage-stage embryos is higher compared to in vitro-produced embryos. Hum Reprod. 2017;32:2348–57.

    PubMed  Google Scholar 

  33. Spits C, Guzman L, Mertzanidou A, Jacobs K, Ortega-Hrepich C, Gilchrist RB, et al. Chromosome constitution of human embryos generated after in vitro maturation including 3-isobutyl-1-methylxanthine in the oocyte collection medium. Hum Reprod. 2015;30:653–63.

    CAS  PubMed  Google Scholar 

  34. Kuhtz J, Romero S, De Vos M, Smitz J, Haaf T, Anckaert E. Human in vitro oocyte maturation is not associated with increased imprinting error rates at LIT1, SNRPN, PEG3 and GTL2. Hum Reprod. 2014;29:1995–2005.

    CAS  PubMed  Google Scholar 

  35. Saenz-de-Juano MD, Ivanova E, Romero S, Lolicato F, Sánchez F, Van Ranst H, et al. DNA methylation and mRNA expression of imprinted genes in blastocysts derived from an improved in vitro maturation method for oocytes from small antral follicles in polycystic ovary syndrome patients. Hum Reprod. 2019;34:1640–9.

    CAS  PubMed  Google Scholar 

  36. Mostinckx L, Segers I, Belva F, Buyl R, Santos-Ribeiro S, Blockeel C, et al. Obstetric and neonatal outcome of ART in patients with polycystic ovary syndrome: IVM of oocytes versus controlled ovarian stimulation. Hum Reprod. 2019;34:1595–607.

    CAS  PubMed  Google Scholar 

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Acknowledgements

The authors would like to thank the full medical and laboratory staff of Brussels IVF for their continuous efforts to maintain and improve the day-to-day infertility care with a special shout-out to our IVM team members: Dr. Caroline Roelens, Dr. Shari Mackens, Dr. Liese Boudry, Dr. Valerie Uvin, Dr. Analisa Racca, Dr Emma Goyens, Griet Meersdom, Griet Eylenbosch, Miranda Verdoodt, Yurda Sert, Yves Guns, Ahmed Afennas, Liesbeth Praet, Emiel De Vroede, and Danijel Jankovic.

Funding

The study was funded by Brussels IVF, UZ-Brussel.

Author information

Authors and Affiliations

  1. Brussels IVF, Universitair Ziekenhuis Brussel, Vrije Universiteit Brussel, Brussels, Belgium

    Ingrid Segers, Lisbet Van Landuyt, Maria-Petra Agius, Linde Mostinckx, Celine Schoemans, Koen Wouters, Neelke De Munck & Michel De Vos

  2. Department of Obstetrics and Gynecology, Mater Dei Hospital, Msida, Malta

    Maria-Petra Agius

  3. Support for Quantitative and Qualitative Research, Vrije Universiteit Brussel, Brussels, Belgium

    Wilfried Cools

  4. GRAD Research Group, Vrije Universiteit Brussel, Brussels, Belgium

    Neelke De Munck

  5. Follicle Biology Laboratory, Vrije Universiteit Brussel, Brussels, Belgium

    Michel De Vos

Authors
  1. Ingrid Segers
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  2. Lisbet Van Landuyt
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  3. Maria-Petra Agius
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  4. Linde Mostinckx
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  6. Celine Schoemans
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  7. Koen Wouters
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  8. Neelke De Munck
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  9. Michel De Vos
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Contributions

IS and MDV were responsible for the concept of the manuscript, IS and MPA were responsible for data retrieval, IS was responsible for data analysis and the initial draft of the article, and WC performed statistical analysis. All authors contributed to the interpretation of the data and editing of the manuscript, critically reviewed the content, and approved the final version.

Corresponding author

Correspondence to Ingrid Segers.

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Ethical approval

Ethical approval for this study was granted by the institutional Ethics Committee of UZ-Brussel (BUN: 1432022000283).

Conflict of interest

The authors declare no competing interests.

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Segers, I., Van Landuyt, L., Agius, MP. et al. Insights gained from a cleavage-stage embryo freeze-all policy in in vitro maturation cycles. J Assist Reprod Genet (2025). https://doi.org/10.1007/s10815-025-03469-7

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