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Blastomere biopsy for PGD delays embryo compaction and blastulation: a time-lapse microscopic analysis

  • Assisted Reproduction Technologies
  • Published:
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Abstract

Purpose

The purpose of the study was to explore the effect of blastomere biopsy for preimplantation genetic diagnosis (PGD) on the embryos’ dynamics, further cleavage, development, and implantation.

Methods

The study group included 366 embryos from all PGD treatments (September 2012 to June 2014) cultured in the EmbryoScope™ time-lapse monitoring system. The control group included all intracytoplasmic sperm injection (ICSI) embryos cultured in EmbryoScope™ until day 5 during the same time period (385 embryos). Time points of key embryonic events were analyzed with an EmbryoViewer™.

Results

Most (88 %) of the embryos were biopsied at ≥8 cells. These results summarize the further dynamic development of the largest cohort of biopsied embryos and demonstrate that blastomere biopsy of cleavage-stage embryos significantly delayed compaction and blastulation compared to the control non-biopsied embryos. This delay in preimplanation developmental events also affected postimplantation development as observed when the dynamics of non-implanted embryos (known implantation data (KID) negative) were compared to those of implanted embryos (KID positive).

Conclusion

Analysis of morphokinetic parameters enabled us to explore how blastomere biopsy interferes with the dynamic sequence of developmental events. Our results show that biopsy delays the compaction and the blastulation of the embryos, leading to a decrease in implantation.

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References

  1. Gutierrez-Mateo C, Sanchez-Garcia JF, Fischer J, Tormasi S, Cohen J, Munne S, et al. Preimplantation genetic diagnosis of single-gene disorders: experience with more than 200 cycles conducted by a reference laboratory in the United States. Fertil Steril. 2009;92(5):1544–56.

    Article  CAS  PubMed  Google Scholar 

  2. Simpson JL. Preimplantation genetic diagnosis to improve pregnancy outcomes in subfertility. Best Pract Res Clin Obstet Gynaecol. 2012;26(6):805–15.

    Article  PubMed  Google Scholar 

  3. Adler A, Lee HL, McCulloh DH, Ampeloquio E, Clarke-Williams M, Wertz BH, et al. Blastocyst culture selects for euploid embryos: comparison of blastomere and trophectoderm biopsies. Reprod Biomed Online. 2014;28(4):485–91.

    Article  PubMed  Google Scholar 

  4. Scott Jr 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(3):624–30.

    Article  PubMed  Google Scholar 

  5. Scott KL, Hong KH, Scott Jr RT. Selecting the optimal time to perform biopsy for preimplantation genetic testing. Fertil Steril. 2013;100(3):608–14.

    Article  PubMed  Google Scholar 

  6. Barbash-Hazan S, Frumkin T, Malcov M, Yaron Y, Cohen T, Azem F, et al. Preimplantation aneuploid embryos undergo self-correction in correlation with their developmental potential. Fertil Steril. 2009;92(3):890–6.

    Article  PubMed  Google Scholar 

  7. Xu K, Montag M. New perspectives on embryo biopsy: not how, but when and why? Semin Reprod Med. 2012;30(4):259–66.

    Article  PubMed  Google Scholar 

  8. Cimadomo D, Capalbo A, Ubaldi FM, Scarica C, Palagiano A, Canipari R, et al. The impact of biopsy on human embryo developmental potential during preimplantation genetic diagnosis. Biomed Res Int. 2016;2016:7193075.

    Article  PubMed  PubMed Central  Google Scholar 

  9. De Rycke M, Belva F, Goossens V, Moutou C, SenGupta SB, Traeger-Synodinos J, et al. ESHRE PGD Consortium data collection XIII: cycles from January to December 2010 with pregnancy follow-up to October 2011. Hum Reprod (Oxford, England). 2015;30(8):1763–89.

    Article  Google Scholar 

  10. Shapiro BS, Daneshmand ST, Garner FC, Aguirre M, Hudson C. Freeze-all at the blastocyst or bipronuclear stage: a randomized clinical trial. Fertil Steril. 2015;104(5):1138–44.

    Article  PubMed  Google Scholar 

  11. Mastenbroek S, Twisk M, van der Veen F, Repping S. Preimplantation genetic screening: a systematic review and meta-analysis of RCTs. Hum Reprod Update. 2011;17(4):454–66.

    Article  CAS  PubMed  Google Scholar 

  12. Ajduk A, Zernicka-Goetz M. Quality control of embryo development. Mol Asp Med. 2013;34(5):903–18.

    Article  Google Scholar 

  13. Ugajin T, Terada Y, Hasegawa H, Velayo CL, Nabeshima H, Yaegashi N. Aberrant behavior of mouse embryo development after blastomere biopsy as observed through time-lapse cinematography. Fertil Steril. 2010;93(8):2723–8.

    Article  PubMed  Google Scholar 

  14. Duncan FE, Stein P, Williams CJ, Schultz RM. The effect of blastomere biopsy on preimplantation mouse embryo development and global gene expression. Fertil Steril. 2009;91(4 Suppl):1462–5.

    Article  PubMed  Google Scholar 

  15. Tarin JJ, Conaghan J, Winston RM, Handyside AH. Human embryo biopsy on the 2nd day after insemination for preimplantation diagnosis: removal of a quarter of embryo retards cleavage. Fertil Steril. 1992;58(5):970–6.

    Article  CAS  PubMed  Google Scholar 

  16. Chawla M, Fakih M, Shunnar A, Bayram A, Hellani A, Perumal V, et al. Morphokinetic analysis of cleavage stage embryos and its relationship to aneuploidy in a retrospective time-lapse imaging study. J Assist Reprod Genet. 2015;32(1):69–75.

    Article  PubMed  Google Scholar 

  17. Meseguer M, Herrero J, Tejera A, Hilligsoe KM, Ramsing NB, Remohi J. The use of morphokinetics as a predictor of embryo implantation. Hum Reprod (Oxford, England). 2011;26(10):2658–71.

    Article  Google Scholar 

  18. Rubio I, Galan A, Larreategui Z, Ayerdi F, Bellver J, Herrero J, et al. Clinical validation of embryo culture and selection by morphokinetic analysis: a randomized, controlled trial of the EmbryoScope. Fertil Steril. 2014;102(5):1287–94.e5.

    Article  PubMed  Google Scholar 

  19. Meseguer M, Rubio I, Cruz M, Basile N, Marcos J, Requena A. Embryo incubation and selection in a time-lapse monitoring system improves pregnancy outcome compared with a standard incubator: a retrospective cohort study. Fertil Steril. 2012;98(6):1481–9.e10.

    Article  PubMed  Google Scholar 

  20. Herrero J, Tejera A, Albert C, Vidal C, de los Santos MJ, Meseguer M. A time to look back: analysis of morphokinetic characteristics of human embryo development. Fertil Steril. 2013;100(6):1602–9.e1-4.

    Article  PubMed  Google Scholar 

  21. Chamayou S, Patrizio P, Storaci G, Tomaselli V, Alecci C, Ragolia C, et al. The use of morphokinetic parameters to select all embryos with full capacity to implant. J Assist Reprod Genet. 2013;30(5):703–10.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Herrero J, Meseguer M. Selection of high potential embryos using time-lapse imaging: the era of morphokinetics. Fertil Steril. 2013;99(4):1030–4.

    Article  PubMed  Google Scholar 

  23. Wu L, Han W, Zhang X, Wang J, Liu W, Xiong S, et al. A retrospective analysis of morphokinetic parameters according to the implantation outcome of IVF treatment. Eur J Obstet Gynecol Reprod Biol. 2016;197:186–90.

    Article  PubMed  Google Scholar 

  24. Park H, Bergh C, Selleskog U, Thurin-Kjellberg A, Lundin K. No benefit of culturing embryos in a closed system compared with a conventional incubator in terms of number of good quality embryos: results from an RCT. Hum Reprod (Oxford, England). 2015;30(2):268–75.

    Article  CAS  Google Scholar 

  25. Mizobe Y, Oya N, Iwakiri R, Yoshida N, Sato Y, Miyoshi K et al. Effects of early cleavage patterns of human embryos on subsequent in vitro development and implantation. Fertil Steril. 2016.

  26. Yang Z, Zhang J, Salem SA, Liu X, Kuang Y, Salem RD, et al. Selection of competent blastocysts for transfer by combining time-lapse monitoring and array CGH testing for patients undergoing preimplantation genetic screening: a prospective study with sibling oocytes. BMC Med Genet. 2014;7:38.

    Google Scholar 

  27. Rienzi L, Capalbo A, Stoppa M, Romano S, Maggiulli R, Albricci L, et al. No evidence of association between blastocyst aneuploidy and morphokinetic assessment in a selected population of poor-prognosis patients: a longitudinal cohort study. Reprod Biomed Online. 2015;30(1):57–66.

    Article  CAS  PubMed  Google Scholar 

  28. Kaser DJ, Racowsky C. Clinical outcomes following selection of human preimplantation embryos with time-lapse monitoring: a systematic review. Hum Reprod Update. 2014;20(5):617–31.

    Article  PubMed  Google Scholar 

  29. Armstrong S, Arroll N, Cree LM, Jordan V, Farquhar C. Time-lapse systems for embryo incubation and assessment in assisted reproduction. Cochrane Database Syst Rev. 2015;2:Cd011320.

    Google Scholar 

  30. Kirkegaard K, Hindkjaer JJ, Ingerslev HJ. Human embryonic development after blastomere removal: a time-lapse analysis. Human Reprod (Oxford, England). 2012;27(1):97–105.

    Article  Google Scholar 

  31. Cruz M, Garrido N, Herrero J, Perez-Cano I, Munoz M, Meseguer M. Timing of cell division in human cleavage-stage embryos is linked with blastocyst formation and quality. Reprod Biomed Online. 2012;25(4):371–81.

    Article  PubMed  Google Scholar 

  32. Staessen C, Verpoest W, Donoso P, Haentjens P, Van der Elst J, Liebaers I, et al. Preimplantation genetic screening does not improve delivery rate in women under the age of 36 following single-embryo transfer. Hum Reprod (Oxford, England). 2008;23(12):2818–25.

    Article  CAS  Google Scholar 

  33. Debrock S, Melotte C, Spiessens C, Peeraer K, Vanneste E, Meeuwis L, et al. Preimplantation genetic screening for aneuploidy of embryos after in vitro fertilization in women aged at least 35 years: a prospective randomized trial. Fertil Steril. 2010;93(2):364–73.

    Article  PubMed  Google Scholar 

  34. Hardarson T, Hanson C, Lundin K, Hillensjo T, Nilsson L, Stevic J, et al. Preimplantation genetic screening in women of advanced maternal age caused a decrease in clinical pregnancy rate: a randomized controlled trial. Hum Reprod (Oxford, England). 2008;23(12):2806–12.

    Article  CAS  Google Scholar 

  35. Pratt HP, Ziomek CA, Reeve WJ, Johnson MH. Compaction of the mouse embryo: an analysis of its components. J Embryol Exp Morphol. 1982;70:113–32.

    CAS  PubMed  Google Scholar 

  36. Clayton L, Stinchcombe SV, Johnson MH. Cell surface localisation and stability of uvomorulin during early mouse development. Zygote (Cambridge, England). 1993;1(4):333–44.

    Article  CAS  Google Scholar 

  37. Sefton M, Johnson MH, Clayton L, McConnell JM. Experimental manipulations of compaction and their effects on the phosphorylation of uvomorulin. Mol Reprod Dev. 1996;44(1):77–87.

    Article  CAS  PubMed  Google Scholar 

  38. Pey R, Vial C, Schatten G, Hafner M. Increase of intracellular Ca2+ and relocation of E-cadherin during experimental decompaction of mouse embryos. Proc Natl Acad Sci U S A. 1998;95(22):12977–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Rink K, Delacretaz G, Salathe RP, Senn A, Nocera D, Germond M, et al. Non-contact microdrilling of mouse zona pellucida with an objective-delivered 1.48-microns diode laser. Lasers Surg Med. 1996;18(1):52–62.

    Article  CAS  PubMed  Google Scholar 

  40. Taylor TH, Gilchrist JW, Hallowell SV, Hanshew KK, Orris JJ, Glassner MJ, et al. The effects of different laser pulse lengths on the embryo biopsy procedure and embryo development to the blastocyst stage. J Assist Reprod Genet. 2010;27(11):663–7.

    Article  PubMed  PubMed Central  Google Scholar 

  41. Malter HE, Cohen J. Blastocyst formation and hatching in vitro following zona drilling of mouse and human embryos. Gamete Res. 1989;24(1):67–80.

    Article  CAS  PubMed  Google Scholar 

  42. Cohen J, Feldberg D. Effects of the size and number of zona pellucida openings on hatching and trophoblast outgrowth in the mouse embryo. Mol Reprod Dev. 1991;30(1):70–8.

    Article  CAS  PubMed  Google Scholar 

  43. Schmoll F, Schneider H, Montag M, Wimmers K, Rink K, Schellander K. Effects of different laser-drilled openings in the zona pellucida on hatching of in vitro-produced cattle blastocysts. Fertil Steril. 2003;80 Suppl 2:714–9.

    Article  PubMed  Google Scholar 

  44. Dahdouh EM, Balayla J, Audibert F, Wilson RD, Audibert F, Brock JA, et al. Technical update: preimplantation genetic diagnosis and screening. J Obstet Gynaecol Canada. 2015;37(5):451–63.

    Article  Google Scholar 

  45. Kokkali G, Traeger-Synodinos J, Vrettou C, Stavrou D, Jones GM, Cram DS, et al. Blastocyst biopsy versus cleavage stage biopsy and blastocyst transfer for preimplantation genetic diagnosis of beta-thalassaemia: a pilot study. Hum Reprod (Oxford, England). 2007;22(5):1443–9.

    Article  CAS  Google Scholar 

  46. Gross L. Built-in timer delays differentiation. PLoS Biol. 2012;10(1):e1001254.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

Embryologists Ariela Carmon, Shiri Asaf, Roni Rahav, and Alina Shpitz are thanked for embryo assessment and documentation. Dana Hadar Fruchter, statistical advisor, is thanked for statistical analysis; Sharon Cooper, statistical advisor, is thanked for descriptive statistics, Esther Eshkol for editorial assistance.

Author information

Authors and Affiliations

  1. IVF Lab and Wolfe PGD-Stem Cell Lab, Racine IVF Unit, Lis Maternity Hospital, Tel-Aviv Sourasky Medical Center, 6 Weizman Street, Tel Aviv, Israel

    Liron Bar-El, Yael Kalma, Mira Malcov, Tamar Schwartz, Shaul Raviv, Tania Cohen, Hadar Amir, Yoni Cohen, Adi Reches, Ami Amit & Dalit Ben-Yosef

  2. Department of Cell Biology and Development, Sackler Faculty of Medicine, Tel-Aviv University, Tel Aviv, Israel

    Dalit Ben-Yosef

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  1. Liron Bar-El
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  2. Yael Kalma
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  8. Yoni Cohen
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Corresponding author

Correspondence to Dalit Ben-Yosef.

Additional information

Capsule Morphokinetic analysis demonstrated that blastomere biopsy interferes with the dynamic sequence of developmental events, delaying compaction and blastulation of the embryos and leading to a decrease in implantation rate.

This work was performed in partial fulfillment of the M.D. thesis requirements of the Sackler Faculty of Medicine, Tel Aviv University.

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Bar-El, L., Kalma, Y., Malcov, M. et al. Blastomere biopsy for PGD delays embryo compaction and blastulation: a time-lapse microscopic analysis. J Assist Reprod Genet 33, 1449–1457 (2016). https://doi.org/10.1007/s10815-016-0813-2

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  • DOI: https://doi.org/10.1007/s10815-016-0813-2

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