Skip to main content
Log in

Accumulation of Cleavage-Stage Embryos by Vitrification may Compromise Embryonic Developmental Potential in Preimplantation Genetic Testing

  • Embryology: Original Article
  • Published:
Reproductive Sciences Aims and scope Submit manuscript

Abstract

It was suggested that the embryo pooling was an alternative for patients with insufficient number of embryos for preimplantation genetic testing (PGT) in a single ovarian stimulation cycle. However, limited study noticed whether it is an efficient strategy to pool cleavage-stage embryos by vitrification. This study included 71 cycles with vitrified-warmed and fresh embryos simultaneously for PGT between May 2016 and May 2021. The embryos from the same patients were split into two groups based on the origin: warming group and fresh group. Embryo development, sequencing results, clinical and neonatal outcomes were compared. The results showed that the rate of high-quality embryos in the warming group was significantly higher than that in the fresh group (64.53% versus 52.61%, P = 0.011); however, the available blastocyst rate in this group was significantly lower than that in the fresh group (47.29% versus 57.83%, P = 0.026). There were 96 and 144 blastocysts that underwent trophectoderm (TE) biopsy in warming and fresh groups, respectively. The high-quality blastocyst rate was significantly lower in the warming group compared to the fresh group (57.29% versus 70.14%, P = 0.041). The rates of genetic transferable blastocyst were comparable between the two groups (P = 0.956). There were no statistical differences in terms of embryo implantation, clinical pregnancy, miscarriage rates, and neonatal outcomes between the two groups. In conclusion, this study demonstrated that the cleavage-stage embryo pooling strategy might be unfavorable for the maintenance of embryonic development potential. If not necessary, it is not recommended to pool cleavage-stage embryos for PGT.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Handyside AH, Kontogianni EH, Hardy K, Winston RM. Pregnancies from biopsied. human preimplantation embryos sexed by Y-specific DNA amplification. Nature. 1990;344:768–70. https://doi.org/10.1038/344768a0.

    Article  CAS  PubMed  Google Scholar 

  2. Scott KL, Hong KH, Scott RT. Selecting the optimal time to perform biopsy for preimplantation genetic testing. Fertil Steril. 2013;100:608–14. https://doi.org/10.1016/j.fertnstert.2013.07.004.

    Article  PubMed  Google Scholar 

  3. Scott RT, Upham KM, Forman EJ, Zhao T, Treff NR. Cleavage-stage biopsy significantly impairs human embryonic implantation potential while blastocyst biopsy does not: a randomized and paired clinical trial. Fertil Steril. 2013;100:624–30. https://doi.org/10.1016/j.fertnstert.2013.04.039.

    Article  PubMed  Google Scholar 

  4. Xiong S, Liu WW, Wang J, Liu JX, Gao Y, Wu LH, et al. Trophectoderm biopsy protocols may impact the rate of mosaic blastocysts in cycles with pre-implantation genetic testing for aneuploidy. J Assist Reprod Genet. 2021; 04. https://doi.org/10.1007/s10815-021-02137-w.

  5. Chamayou S, Sicali M, Alecci C, Ragolia C, Liprino A, Nibali D, et al. The accumulation of vitrified oocytes is a strategy to increase the number of euploid available blastocysts for transfer after preimplantation genetic testing. J Assist Reprod Genet. 2017;34:479–86. https://doi.org/10.1007/s10815-016-0868-0.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Xiaokun Hu, Chenhui D, DuoDuo Z, Wen Z, Jing W, Yanhong Z, et al. Embryo pooling: a promising strategy for managing insufficient number of embryos in preimplantation genetic diagnosis. Gynecol Endocrinol. 2017;33:867–71. https://doi.org/10.1080/09513590.2017.1347778.

    Article  Google Scholar 

  7. Chatziparasidou A, Nijs M, Moisidou M, Chara O, Ioakeimidou C, Pappas C, et al. Accumulation of oocytes and/or embryos by vitrification: a new strategy for managing poor responder patients undergoing preimplantation diagnosis. F1000Res. 2013;2:240. https://doi.org/10.12688/f1000research.2-240.v2.

    Article  PubMed  Google Scholar 

  8. Orris JJ, Taylor TH, Gilchrist JW, Hallowell SV, Glassner MJ, Wininger JD. The utility of embryo banking in order to increase the number of embryos available for preimplantation genetic screening in advanced maternal age patients. J Assist Reprod Genet. 2010;27:729–33. https://doi.org/10.1007/s10815-010-9474-8.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Mansoori GA. Kinetics of water loss from cells at subzero centigrade temperatures. Cryobiology. 1975;12:34–45.

    Article  CAS  Google Scholar 

  10. Kleinhans FW, Mazur P. Comparison of actual vs. synthesized ternary phase diagrams for solutes of cryobiological interest. Cryobiology. 2007;54:212–22. https://doi.org/10.1016/j.cryobiol.2007.01.007.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Pinborg A, Henningsen AA, Loft A, Malchau SS, Forman J, Andersen AN. Large baby syndrome in singletons born after frozen embryo transfer (FET): is it due to maternal factors or the cryotechnique? Hum Reprod. 2014;29:618–27. https://doi.org/10.1093/humrep/det440.

    Article  CAS  PubMed  Google Scholar 

  12. Stanger J, Wong J, Conceicao J, Yovich J. Vitrification of human embryos previously cryostored by either slow freezing or vitrification results in high pregnancy rates. Reprod Biomed Online. 2012;24:314–20. https://doi.org/10.1016/j.rbmo.2011.11.013.

    Article  CAS  PubMed  Google Scholar 

  13. Taylor TH, Patrick JL, Gitlin SA, Michael Wilson J, Crain JL, Griffin DK, et al. Outcomes of blastocysts biopsied and vitrified once versus those cryopreserved twice for euploid blastocyst transfer. Reprod Biomed Online. 2014;29:59–64. https://doi.org/10.1016/j.rbmo.2014.03.001.

    Article  PubMed  Google Scholar 

  14. Cimadomo D, Rienzi L, Romanelli V, Alviggi E, Levi-Setti PE, Albani E, et al. Inconclusive chromosomal assessment after blastocyst biopsy: prevalence, causative factors and outcomes after re-biopsy and re-vitrification A multicenter experience. Hum Reprod. 2018;33:1839–46. https://doi.org/10.1093/humrep/dey282.

    Article  PubMed  Google Scholar 

  15. Wang M, Jiang JH, Xi QS, Li D, Ren XL, Li Z, et al. Repeated cryopreservation process impairs embryo implantation potential but does not affect neonatal outcomes. Reprod Biomed Online. 2021;42:75–82. https://doi.org/10.1016/j.rbmo.2020.11.007.

    Article  CAS  PubMed  Google Scholar 

  16. Zheng XY, Chen Y, Yan J, Wu YQ, Zhuang XJ, Lin SL, et al. Effect of repeated cryopreservation on human embryo developmental potential. Reprod Biomed Online. 2017;35:627–32. https://doi.org/10.1016/j.rbmo.2017.08.016.

    Article  PubMed  Google Scholar 

  17. Ding X, Yang J, Li L, Yang N, Lan L, Huang G, Ye H. Fertility outcomes in women after controlled ovarian stimulation with gonadotropin releasing hormone agonist long protocol: fresh versus frozen embryo transfer. BMC Pregnancy Childbirth. 2021;21:207. https://doi.org/10.1186/s12884-021-03698-5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Jingyu Li, Shun X, Yanhua Z, Chong Li, Wei H, Guoning H. Effect of the re-vitrification of embryos at different stages on embryonic developmental potential. Front Endocrinol. 2021;12:653310. https://doi.org/10.3389/fendo.2021.653310.

    Article  Google Scholar 

  19. Xiong S, Han W, Liu JX, Zhang XD, Liu WW, Liu H, et al. Effects of cumulus cells removal after 6 h co-incubation of gametes on the outcomes of human IVF. J Assist Reprod Genet. 2011;28:1205–11. https://doi.org/10.1007/s10815-011-9630-9.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Gardner DK, Lane M. Culture and selection of viable blastocysts: a feasible proposition for human IVF? Hum Reprod Update. 1997;3:367–82. https://doi.org/10.1093/humupd/3.4.367.

    Article  CAS  PubMed  Google Scholar 

  21. Xiong S, Liu JX, Gao Y, Liu WW, Wu LH, Han W, et al. Shortened equilibration time can compromise clinical outcomes in human embryo vitrification. Hum Fertil (Camb). 2016;19:114–9. https://doi.org/10.1080/14647273.2016.1186848.

    Article  PubMed  Google Scholar 

  22. Capalbo A, Rienzi L, Cimadomo D, Maggiulli R, Elliott T, Wright G, et al. Correlation between standard blastocyst morphology, euploidy and implantation: an observational study in two centers involving 956 screened blastocysts. Hum Reprod. 2014;29:1173–81. https://doi.org/10.1093/humrep/deu033.

    Article  PubMed  Google Scholar 

  23. Hu XK, Ding CH, Zhang DD, Zhou W, Wang J, Zeng YH, et al. Embryo pooling: a promising strategy for managing insufficient number of embryos in preimplantation genetic diagnosis. Gynecol Endocrinol. 2017;33:867–71. https://doi.org/10.1080/09513590.2017.1347778.

    Article  PubMed  Google Scholar 

  24. Wilding M, Terribile M, Parisi I, Nargund G. Thaw, biopsy and refreeze strategy for PGT-A on previously cryopreserved embryos. Facts Views Vis Obgyn. 2019;11:223–7.

    CAS  PubMed  Google Scholar 

  25. Anick DV, Lisbet VL, Martine DR, Pieter V, Greta V, Andrea B, et al. Multiple vitrification-warming and biopsy procedures on human embryos: clinical outcome and neonatal follow-up of children. Hum Reprod. 2020;35:2488–96. https://doi.org/10.1093/humrep/deaa236.

    Article  Google Scholar 

  26. Lan Yu, Jia C, Lan Y, Song R, Zhou L, Li Y, et al. Analysis of embryo intactness and developmental potential following slow freezing and vitrification. Syst Biol Reprod Med. 2017;63:285–93. https://doi.org/10.1080/19396368.2017.1362060.

    Article  CAS  PubMed  Google Scholar 

  27. Vanderzwalmen P, Ectors F, Grobet L, Prapas Y, Panagiotidis Y, Vanderzwalmen S, et al. Aseptic vitrification of blastocysts from infertile patients, egg donors and after IVM. Reprod Biomed Online. 2009;19:700–7. https://doi.org/10.1016/j.rbmo.2009.09.011.

    Article  CAS  PubMed  Google Scholar 

  28. Kader A, Choi A, Sharma RK, Falcone T, Agarwal A. Effect of varying equilibration time in a two-step vitrification method on the post-warming DNA integrity of mouse blastocysts. Fertil Steril. 2010;93:2640–5. https://doi.org/10.1016/j.fertnstert.2009.07.991.

    Article  PubMed  Google Scholar 

  29. Chatzimeletiou K, Morrison EE, Vanderzwalmen P, Prapas N, Prapas Y, Basil C, et al. Cytoskeletal analysis of human blastocysts by confocal laser scanning microscopy following vitrification. Hum Reprod. 2012;27:106–13. https://doi.org/10.1093/humrep/der344.

    Article  PubMed  Google Scholar 

  30. Zhang SP, Tan K, Gong F, Gu YF, Tan YQ, Lu CF, et al. Blastocysts can be rebiopsied for preimplantation genetic diagnosis and screening. Fertil Steril. 2014;102:1641–5. https://doi.org/10.1016/j.fertnstert.2014.09.018.

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

Our thanks should be given to all the embryologists and biologists in charge of embryo biopsy, cryopreservation, and NGS analysis.

Funding

This work was supported by the Chongqing Health Committee (grant number: 2019MSXM067, 2020MSXM086 and 2021MSXM108).

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Wei Han or Guoning Huang.

Ethics declarations

Competing Interests

The authors declare no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Shun Xiong and Xiangwei Hao are contributed equally to the research and preparation of the manuscript.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Xiong, S., Hao, X., Gao, Y. et al. Accumulation of Cleavage-Stage Embryos by Vitrification may Compromise Embryonic Developmental Potential in Preimplantation Genetic Testing. Reprod. Sci. 29, 1930–1938 (2022). https://doi.org/10.1007/s43032-022-00880-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s43032-022-00880-8

Keywords

Navigation