Comparison of two protocols of blastocyst biopsy submitted to preimplantation genetic testing for aneuploidies: a randomized controlled trial

  • Haibin Zhao
  • Wenrong Tao
  • Mei Li
  • Hui Liu
  • Keliang Wu
  • Shuiying MaEmail author
Gynecologic Endocrinology and Reproductive Medicine



To compare the effectiveness of two protocols of blastocyst biopsy submitted to preimplantation genetic testing for aneuploidies (PGT-A).


This is a randomized controlled trial of a cohort of 221 patients undergoing PGT-A. 106 female patients aged ≤ 40 years with no less than 8 mature oocytes retrieved and ≥ 3 good-quality embryos on day 3 were randomly assigned to the day-3 hatching-based TE biopsy. The remaining 115 females aged ≤ 40 years with ≥ 8 MII oocytes obtained and no less than 3 high-quality embryos on day 3 were assigned to the TE biopsy without hatching group (also called the new biopsy group). The primary outcome was measured by a live birth after the first embryo transfer.


The live birth rate did not differ significantly between the two groups (50.00% vs. 59.26%, P > 0.05, OR 1.46; 95% CI 0.78–2.70). There was no significant between-group difference in the rates of implantation, clinical pregnancy, and miscarriage. However, the frozen blastocyst rate was significantly lower in the day-3 hatching-based TE biopsy compared with the new biopsy group (47.54% vs. 53.96%, P < 0.05, OR 1.29; 95% CI 1.08–1.56).


Our study provides strong evidence that the new blastocyst biopsy method exhibits advantages over day-3 hatching-based TE biopsy method. Using this method, we were able to obtain more blastocysts to perform trophectoderm biopsy in patients subjected to PGT-A.


Preimplantation genetic testing for aneuploidies (PGT-A) Blastocyst biopsy Embryonic aneuploidy Next-generation sequencing Frozen embryo transfer 


Author contributions

LH: data management/analysis, manuscript writing/editing, ZHB: data management/analysis, manuscript writing/editing, YGL: data collection, LM: manuscript editing, MSY: manuscript editing, ZHZ: data collection, WKL: study design, manuscript writing/editing.


This study is funded by National Natural Science Foundation of China (no. 81601256) and Merck Serono China Research Fund for Fertility.

Compliance with ethical standards

Conflict of interest

All the authors declare no conflict of interest with respect to the authorship and/or publication of this article.

Ethical approval

All procedures performed in this study involving human participants were in accordance with the ethical standards of the Reproductive Medicine, Shandong University Research Ethics Committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in this study.


  1. 1.
    Kuliev A, Verlinsky Y (2004) Meiotic and mitotic nondisjunction: lessons from preimplantation genetic diagnosis. Hum Reprod Update 10:401–407. CrossRefGoogle Scholar
  2. 2.
    Pellestor F, Andreo B, Anahory T, Hamamah S (2006) The occurrence of aneuploidy in human: lessons from the cytogenetic studies of human oocytes. Eur J Med Genet 49:103–116. CrossRefGoogle Scholar
  3. 3.
    Centers for Disease Control and Prevention (2009) National summary report. Assisted reproductive technology (ART) report. CDC, AtlantaGoogle Scholar
  4. 4.
    Hassold T, Hunt P (2001) To err (meiotically) is human: the genesis of human aneuploidy. Nat Rev 2:280–291. CrossRefGoogle Scholar
  5. 5.
    Forman EJ, Tao X, Ferry KM, Taylor D, Treff NR, Scott RT (2012) Single embryo transfer with comprehensive chromosome screening results in improved ongoing pregnancy rates and decreased miscarriage rates. Hum Reprod 27:1217–1222. CrossRefGoogle Scholar
  6. 6.
    Fragouli E, Wells D (2011) Aneuploidy in the human blastocyst. Cytogenet Genome Res 133:149–159. CrossRefGoogle Scholar
  7. 7.
    Hardarson T, Hanson C, Lundin K, Hillensjo T, Nilsson L, Stevic J, Reismer E, Borg K, Wikland M, Bergh C (2008) Preimplantation genetic screening in women of advanced maternal age caused a decrease in clinical pregnancy rate: a randomized controlled trial. Hum Reprod 23:2806–2812. CrossRefGoogle Scholar
  8. 8.
    Mastenbroek S, Twisk M, Van Echten-Arends J, Sikkema-Raddatz B, Korevaar JC, Verhoeve HR, Vogel NE, Arts EG, De Vries JW, Bossuyt PM, Buys CH, Heineman MJ, Repping S, Van Der Veen F (2007) In vitro fertilization with preimplantation genetic screening. N Engl J Med 357:9–17. CrossRefGoogle Scholar
  9. 9.
    Mastenbroek S, Twisk M, Van Der Veen F, Repping S (2011) Preimplantation genetic screening: a systematic review and meta analysis of RCTs. Hum Reprod Update 17:454–466. CrossRefGoogle Scholar
  10. 10.
    Harton GL, Munne S, Surrey M, Grifo J, Kaplan B, McCulloh DH, Griffin DK, Wells D (2013) Diminished effect of maternal age on implantation after preimplantation genetic diagnosis with array comparative genomic hybridization. Fertil Steril 100:1695–1703. CrossRefGoogle Scholar
  11. 11.
    Schoolcraft WB, Fragouli E, Stevens J, Munne S, Katz-Jaffe MG, Wells D (2010) Clinical application of comprehensive chromosomal screening at the blastocyst stage. Fertil Steril 94:1700–1706. CrossRefGoogle Scholar
  12. 12.
    Scott RT Jr, Upham KM, Forman EJ, Hong KH, Scott KL, Taylor D, Tao X, Treff NR (2013) Blastocyst biopsy with comprehensive chromosome screening and fresh embryo transfer significantly increases in vitro fertilization implantation and delivery rates: a randomized controlled trial. Fertil Steril 100:697–703. CrossRefGoogle Scholar
  13. 13.
    Yang Z, Liu J, Collins GS, Salem SA, Liu X, Lyle SS (2012) Selection of single blastocysts for fresh transfer via standard morphology assessment alone and with array CGH for good prognosis IVF patients: results from a randomized pilot study. Mol Cytogenet 5:24. CrossRefGoogle Scholar
  14. 14.
    McArthur SJ, Leigh D, Marshall JT, de Boer KA, Jansen RP (2005) Pregnancies and live births after trophectoderm biopsy and preimplantation genetic testing of human blastocysts. Fertil Steril 84:1628–1636. CrossRefGoogle Scholar
  15. 15.
    De Boer KA, McArthur S, Murray C, Jansen R (2002) First live birth following blastocyst biopsy and PGD analysis. RBM Online 4:35. Google Scholar
  16. 16.
    De Boer KA, Catt JW, Jansen RP, Leigh D, McArthur S (2004) Moving to blastocyst biopsy for preimplantation genetic diagnosis and single embryo transfer at Sydney IVF. Fertil Steril 82(2):295–298. CrossRefGoogle Scholar
  17. 17.
    Puissant F, Van RM, Barlow P (1987) Embryo scoring as a prognostic tool in IVF treatment. Hum Reprod 2:705–708CrossRefGoogle Scholar
  18. 18.
    Wu KL, Zhao HB, Liu H, Li M, Ma S, Li C, Liu C, Chen ZJ (2014) Day 3 ET, single blastocyst transfer (SBT) or frozen-thawed embryo transfer (FET): which is preferable for high responder patients in IVF/ICSI cycles? J Assist Reprod Genet 31:275–278. CrossRefGoogle Scholar
  19. 19.
    Gardner DK, Lane M (1997) Culture and selection of viable blastocysts: a feasible proposition for human IVF? Hum Reprod Update 3:367–382CrossRefGoogle Scholar
  20. 20.
    Cimadomo D, Capalbo A, Ubaldi FM, Rienzi L (2016) Laser-assisted zona opening and trophectoderm biopsy at the blastocyst stage: a video guide. Curr Trends Clin Embryol 3(2):73–74. CrossRefGoogle Scholar
  21. 21.
    Capalbo A, Rienzi L, Cimadomo D, Maggiulli R, Elliott T, Wright G, Nagy ZP, Ubaldi FM (2014) Correlation between standard blastocyst morphology, euploidy and implantation: an observational study in two centers involving 956 screened blastocysts. Hum Reprod 29(6):1173–1181. CrossRefGoogle Scholar
  22. 22.
    Treff NR, Forman EJ, Scott RT (2013) Next-generation sequencing for preimplantation genetic diagnosis. Fertil Steril 99:e17–e18. CrossRefGoogle Scholar
  23. 23.
    Wells D (2014) Next-generation sequencing: the dawn of a new era for preimplantation genetic diagnostics. Fertil Steril 101:1250–1251. CrossRefGoogle Scholar
  24. 24.
    Mukaida T, Nakamura S, Tomiyama T, Wada S, Kasai M, Takahashi K (2001) Successful birth after transfer of vitrified human blastocysts with use of a cryoloop containerless technique. Fertil Steril 76:618–620CrossRefGoogle Scholar
  25. 25.
    Alfarawati S, Fragouli E, Colls P, Stevens J, Gutierrez-Mateo C, Schoolcraft WB, Katz-Jaffe MG, Wells D (2011) The relationship between blastocyst morphology, chromosomal abnormality, and embryo gender. Fertil Steril 95(2):520–524. CrossRefGoogle Scholar
  26. 26.
    Irani M, Reichman D, Robles A, Melnick A, Davis O, Zaninovic N, Xu K, Rosenwaks Z (2017) Morphologic grading of euploid blastocysts influences implantation and ongoing pregnancy rates. Fertil Steril 107:664–670. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Center for Reproductive Medicine, Shandong Provincial Hospital Affiliated to Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive GeneticsChina The Key laboratory of Reproductive Endocrinology (Shandong University), Ministry of Education Shandong Provincial Key Laboratory of Reproductive MedicineJinanPeople’s Republic of China

Personalised recommendations