Skip to main content
SpringerLink
Log in

Day 5 vs day 6 single euploid blastocyst frozen embryo transfers: which variables do have an impact on the clinical pregnancy rates?

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

Abstract

Objective

To determine which variables affect most the clinical pregnancy rate with positive fetal heartbeat (CPR FHB+) when frozen embryo transfer (FET) cycles are performed with day 5 (D5) or day 6 (D6) euploid blastocysts.

Design and method

A single center retrospective study was performed from March 2017 till February 2021 including all single FET cycles with euploid D5 or D6 blastocysts and transferred in natural cycles (NC) or hormone replacement therapy (HRT) cycles. Trophectoderm (TE) and inner cell mass (ICM) qualities were recorded before biopsy.

Results

A total of 1102 FET cycles were included, 678 with D5 and 424 with D6 blastocysts. Pregnancy rate (PR), clinical PR (CPR), and CPR FHB+ were significantly higher with D5 blastocysts (PR: 70.7% vs 62.0%, OR = 0.68 [0.53–0.89], p = 0.004; CPR: 63.7% vs 54.2%, OR = 0.68 [0.52–0.96], p = 0.002 and CPR FHB+: 57.8% vs 49.8%, OR = 0.72 [0.53–0.96], p = 0.011). However, miscarriage rate (12.5% vs 11.4%, OR = 0.78 [0.48–1.26], p = 0.311) did not differ. From a multivariate logistic regression model, endometrial thickness (OR = 1.11 [1.01–1.22], p = 0.028), patient’s age (OR = 1.03 [1.00–1.05], p = 0.021), BMI (OR = 0.97 [0.94–0.99], p = 0.023), and ICM grade C (OR = 0.23 [0.13–0.43], p < 0.001) were significant in predicting CPR FHB+.

Conclusion

Although clinical outcomes are higher with D5 blastocysts, CPR FHB+ is more affected by endometrial thickness, patient age, BMI, and ICM grade C rather than biopsy day or endometrial preparation protocol.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  1. Capalbo A, Rienzi L, Cimadomo D, Maggiulli R, Elliott T, Wright G, Nagy ZP, Ubaldi FM. Correlation between standard blastocyst morphology, euploidy and implantation: an observational study in two centers involving 956 screened blastocysts. Hum Reprod. 2014;29(6):1173–81. https://doi.org/10.1093/humrep/deu033.

    Article  PubMed  Google Scholar 

  2. Alpha Scientists in Reproductive Medicine and ESHRE Special Interest Group of Embryology. The Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting. Hum Reprod. 2011;26(6):1270–83. https://doi.org/10.1093/humrep/der037.

    Article  Google Scholar 

  3. Tiegs AW, Sun L, Patounakis G, Scott RT. Worth the wait? Day 7 blastocysts have lower euploidy rates but similar sustained implantation rates as day 5 and day 6 blastocysts. Hum Reprod. 2019;34(9):1632–9. https://doi.org/10.1093/humrep/dez138 Erratum in: Hum Reprod. 2019;34(12):2559-2560.

    Article  CAS  PubMed  Google Scholar 

  4. Tannus S, Cohen Y, Henderson S, Al Ma'mari N, Shavit T, Son WY, Dahan MH. Fresh transfer of day 5 slow-growing embryos versus deferred transfer of vitrified, fully expanded day 6 blastocysts: which is the optimal approach? Hum Reprod. 2019;34(1):44–51. https://doi.org/10.1093/humrep/dey351.

    Article  PubMed  Google Scholar 

  5. Shapiro BS, Daneshmand ST, Restrepo H, Garner FC, Aguirre M, Hudson C. Matched-cohort comparison of single-embryo transfers in fresh and frozen-thawed embryo transfer cycles. Fertil Steril. 2013;99(2):389–92. https://doi.org/10.1016/j.fertnstert.2012.09.044.

    Article  PubMed  Google Scholar 

  6. Bourdon M, Pocate-Cheriet K, Finet de Bantel A, Grzegorczyk-Martin V, Amar Hoffet A, Arbo E, Poulain M, Santulli P. day 5 versus day 6 blastocyst transfers: a systematic review and meta-analysis of clinical outcomes. Hum Reprod. 2019;34(10):1948–64. https://doi.org/10.1093/humrep/dez163.

    Article  PubMed  PubMed Central  Google Scholar 

  7. Bakkensen JB, Brady P, Carusi D, Romanski P, Thomas AM, Racowsky C. Association between blastocyst morphology and pregnancy and perinatal outcomes following fresh and cryopreserved embryo transfer. J Assist Reprod Genet. 2019;36(11):2315–24. https://doi.org/10.1007/s10815-019-01580-0.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Roque M, Bedoschi G, Cecchino GN, Esteves SC. Fresh versus frozen blastocyst transfer. Lancet. 2019;394(10205):1227–8. https://doi.org/10.1016/S0140-6736(19)31393-5.

    Article  PubMed  Google Scholar 

  9. Shi Y, Sun Y, Hao C, Zhang H, Wei D, Zhang Y, Zhu Y, Deng X, Qi X, Li H, Ma X, Ren H, Wang Y, Zhang D, Wang B, Liu F, Wu Q, Wang Z, Bai H, et al. Transfer of fresh versus frozen embryos in ovulatory women. N Engl J Med. 2018;378(2):126–36. https://doi.org/10.1056/NEJMoa1705334.

    Article  PubMed  Google Scholar 

  10. Stormlund S, Sopa N, Zedeler A, Bogstad J, Prætorius L, Nielsen HS, Kitlinski ML, Skouby SO, Mikkelsen AL, Spangmose AL, Jeppesen JV, Khatibi A, la Cour FN, Ziebe S, Polyzos NP, Bergh C, Humaidan P, Andersen AN, Løssl K, Pinborg A. Freeze-all versus fresh blastocyst transfer strategy during in vitro fertilization in women with regular menstrual cycles: multicentre randomised controlled trial. BMJ. 2020;370:m2519. https://doi.org/10.1136/bmj.m2519.

    Article  PubMed  PubMed Central  Google Scholar 

  11. Desai N, Ploskonka S, Goodman L, Attaran M, Goldberg JM, Austin C, Falcone T. Delayed blastulation, multinucleation, and expansion grade are independently associated with live-birth rates in frozen blastocyst transfer cycles. Fertil Steril. 2016;106(6):1370–8. https://doi.org/10.1016/j.fertnstert.2016.07.1095.

    Article  PubMed  Google Scholar 

  12. Sciorio R, Thong KJ, Pickering SJ. Increased pregnancy outcome after day 5 versus day 6 transfers of human vitrified-warmed blastocysts. Zygote. 2019;27(5):279–84. https://doi.org/10.1017/S0967199419000273.

    Article  CAS  PubMed  Google Scholar 

  13. Ferreux L, Bourdon M, Sallem A, Santulli P, Barraud-Lange V, Le Foll N, Maignien C, Chapron C, de Ziegler D, Wolf JP, Pocate-Cheriet K. Live birth rate following frozen-thawed blastocyst transfer is higher with blastocysts expanded on day 5 than on day 6. Hum Reprod. 2018;33(3):390–8. https://doi.org/10.1093/humrep/dey004.

    Article  PubMed  Google Scholar 

  14. Sunkara SK, Siozos A, Bolton VN, Khalaf Y, Braude PR, El-Toukhy T. The influence of delayed blastocyst formation on the outcome of frozen-thawed blastocyst transfer: a systematic review and meta-analysis. Hum Reprod. 2010;25(8):1906–15. https://doi.org/10.1093/humrep/deq143.

    Article  PubMed  Google Scholar 

  15. El-Toukhy T, Wharf E, Walavalkar R, Singh A, Bolton V, Khalaf Y, Braude P. Delayed blastocyst development does not influence the outcome of frozen-thawed transfer cycles. BJOG. 2011;118(13):1551–6. https://doi.org/10.1111/j.1471-0528.2011.03101.x.

    Article  CAS  PubMed  Google Scholar 

  16. Reignier A, Lammers J, Barriere P, Freour T. Can time-lapse parameters predict embryo ploidy? A systematic review. Reprod Biomed Online. 2018;36(4):380–7. https://doi.org/10.1016/j.rbmo.2018.01.001.

    Article  PubMed  Google Scholar 

  17. Barash OO, Ivani KA, Willman SP, Rosenbluth EM, Wachs DS, Hinckley MD, Pittenger Reid S, Weckstein LN. Association between growth dynamics, morphological parameters, the chromosomal status of the blastocysts, and clinical outcomes in IVF PGS cycles with single embryo transfer. J Assist Reprod Genet. 2017;34(8):1007–16. https://doi.org/10.1007/s10815-017-0944-0.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Irani M, O'Neill C, Palermo GD, Xu K, Zhang C, Qin X, Zhan Q, Clarke RN, Ye Z, Zaninovic N, Rosenwaks Z. Blastocyst development rate influences implantation and live birth rates of similarly graded euploid blastocysts. Fertil Steril. 2018;110(1):95–102.e1. https://doi.org/10.1016/j.fertnstert.2018.03.032.

    Article  PubMed  Google Scholar 

  19. Kaye L, Will EA, Bartolucci A, Nulsen J, Benadiva C, Engmann L. Pregnancy rates for single embryo transfer (SET) of day 5 and day 6 blastocysts after cryopreservation by vitrification and slow freeze. J Assist Reprod Genet. 2017;34(7):913–9. https://doi.org/10.1007/s10815-017-0940-4.

    Article  PubMed  PubMed Central  Google Scholar 

  20. El-Damen A, Elkhatib I, Bayram A, Arnanz A, Abdala A, Samir S, Lawrenz B, De Munck N, Fatemi HM. Does blastocyst mitochondrial DNA content affect miscarriage rate in patients undergoing single euploid frozen embryo transfer? J Assist Reprod Genet. 2021;38(3):595–604. https://doi.org/10.1007/s10815-020-02050-8.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Greco E, Litwicka K, Arrivi C, Varricchio MT, Caragia A, Greco A, Minasi MG, Fiorentino F. The endometrial preparation for frozen-thawed euploid blastocyst transfer: a prospective randomized trial comparing clinical results from natural modified cycle and exogenous hormone stimulation with GnRH agonist. J Assist Reprod Genet. 2016;33(7):873–84. https://doi.org/10.1007/s10815-016-0736-y.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Ghobara T, Gelbaya TA, Ayeleke RO. Cycle regimens for frozen-thawed embryo transfer. Cochrane Database Syst Rev. 2017;7(7):CD003414. https://doi.org/10.1002/14651858.CD003414.pub3.

    Article  PubMed  Google Scholar 

  23. Glujovsky D, Pesce R, Sueldo C, Quinteiro Retamar AM, Hart RJ, Ciapponi A. Endometrial preparation for women undergoing embryo transfer with frozen embryos or embryos derived from donor oocytes. Cochrane Database Syst Rev. 2020;10:CD006359. https://doi.org/10.1002/14651858.CD006359.pub3 Accessed 19 July 2021.

    Article  PubMed  Google Scholar 

  24. Groenewoud ER, Cantineau AE, Kollen BJ, Macklon NS, Cohlen BJ. What is the optimal means of preparing the endometrium in frozen-thawed embryo transfer cycles? A systematic review and meta-analysis. Hum Reprod Update. 2013;19(5):458–70. https://doi.org/10.1093/humupd/dmt030 Erratum in: Hum Reprod Update. 2017 Mar 1;23(2):255-261.

    Article  PubMed  Google Scholar 

  25. Gardner DK, Schoolcraft WB. In vitro culture of human blastocyst. In: Jansen R, Mortimer D, editors. Towards reproductive certainty: infertility and genetics beyond 1999. Carnforth: Parthenon Press; 1999. p. 378–88.

    Google Scholar 

  26. Kuwayama M. Highly efficient vitrification for cryopreservation of human oocytes and embryos: the Cryotop method. Theriogenology. 2007;67:73–80.

    Article  CAS  PubMed  Google Scholar 

  27. Shu Y, Watt J, Gebhardt J, Dasig J, Appling J, Behr B. The value of fast blastocoele re-expansion in the selection of a viable thawed blastocyst for transfer. Fertil Steril. 2009;91(2):401–6. https://doi.org/10.1016/j.fertnstert.2007.11.083.

    Article  PubMed  Google Scholar 

  28. Fatemi HM, Kyrou D, Bourgain C, Van den Abbeel E, Griesinger G, Devroey P. Cryopreserved-thawed human embryo transfer: spontaneous natural cycle is superior to human chorionic gonadotropin-induced natural cycle. Fertil Steril. 2010;94(6):2054–8. https://doi.org/10.1016/j.fertnstert.2009.11.036.

    Article  CAS  PubMed  Google Scholar 

  29. Davis O, Zaninovic N, Xu K, Rosenwaks Z. Morphologic grading of euploid blastocysts influences implantation and ongoing pregnancy rates. Fertil Steril. 2017;107(3):664–70. https://doi.org/10.1016/j.fertnstert.2016.11.012.

    Article  PubMed  Google Scholar 

  30. Lee CI, Chen CH, Huang CC, Cheng EH, Chen HH, Ho ST, Lin PY, Lee MS, Lee TH. Embryo morphokinetics is potentially associated with clinical outcomes of single-embryo transfers in preimplantation genetic testing for aneuploidy cycles. Reprod BioMed Online. 2019;39(4):569–79. https://doi.org/10.1016/j.rbmo.2019.05.020.

    Article  CAS  PubMed  Google Scholar 

  31. Minasi MG, Colasante A, Riccio T, Ruberti A, Casciani V, Scarselli F, Spinella F, Fiorentino F, Varricchio MT, Greco E. Correlation between aneuploidy, standard morphology evaluation and morphokinetic development in 1730 biopsied blastocysts: a consecutive case series study. Hum Reprod. 2016;31(10):2245–54. https://doi.org/10.1093/humrep/dew183.

    Article  PubMed  Google Scholar 

  32. Park DS, Kim JW, Chang EM, Lee WS, Yoon TK, Lyu SW. Obstetric, neonatal, and clinical outcomes of day 6 vs. day 5 vitrified-warmed blastocyst transfers: retrospective cohort study with propensity score matching. Front Endocrinol (Lausanne). 2020;11:499. https://doi.org/10.3389/fendo.2020.00499.

    Article  Google Scholar 

  33. Ji H, Zhou Y, Cao S, Zhang J, Ling X, Zhao C, Shen R. Effect of embryo developmental stage, morphological grading, and ploidy status on live birth rate in frozen cycles of single blastocyst transfer. Reprod Sci. 2021;28(4):1079–91. https://doi.org/10.1007/s43032-020-00381-6.

    Article  PubMed  Google Scholar 

  34. Li YX, Wang J, Sun TZ, Lv MQ, Ge P, Li HN, Zhou DX. Pregnancy outcomes after day 5 versus day 6 blastocyst-stage embryo transfer: a systematic review and meta-analysis. J Obstet Gynaecol Res. 2020;46(4):595–605. https://doi.org/10.1111/jog.14188.

    Article  PubMed  Google Scholar 

  35. Annan JJ, Gudi A, Bhide P, Shah A, Homburg R. Biochemical pregnancy during assisted conception: a little bit pregnant. J Clin Med Res. 2013;5(4):269–74. https://doi.org/10.4021/jocmr1008w.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Chen L, Diao Z, Wang J, Xu Z, Zhang N, Fang J, Lin F. The effects of the day of trophectoderm biopsy and blastocyst grade on the clinical and neonatal outcomes of preimplantation genetic testing-frozen embryo transfer cycles. Zygote. 2021;28:1–6. https://doi.org/10.1017/S0967199421000435.

    Article  CAS  Google Scholar 

  37. Ai J, Jin L, Zheng Y, Yang P, Huang B, Dong X. The morphology of inner cell mass is the strongest predictor of live birth after a frozen-thawed single embryo transfer. Front Endocrinol (Lausanne). 2021;12:621221. https://doi.org/10.3389/fendo.2021.621221.

    Article  Google Scholar 

  38. Zhao YY, Yu Y, Zhang XW. Overall blastocyst quality, trophectoderm grade, and inner cell mass grade predict pregnancy outcome in euploid blastocyst transfer cycles. Chin Med J. 2018;131(11):1261–7. https://doi.org/10.4103/0366-6999.232808.

    Article  PubMed  PubMed Central  Google Scholar 

  39. Lou H, Li N, Guan Y, Zhang Y, Hao D, Cui S. Association between morphologic grading and implantation rate of euploid blastocyst. J Ovarian Res. 2021;14(1):18. https://doi.org/10.1186/s13048-021-00770-8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Boynukalin FK, Gultomruk M, Cavkaytar S, Turgut E, Findikli N, Serdarogullari M, Coban O, Yarkiner Z, Rubio C, Bahceci M. Parameters impacting the live birth rate per transfer after frozen single euploid blastocyst transfer. PLoS One. 2020;15(1):e0227619. https://doi.org/10.1371/journal.pone.0227619.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Kimelman D, Confino R, Okeigwe I, Lambe-Steinmiller J, Confino E, Shulman LP, Zhang JX, Pavone ME. Assessing the impact of delayed blastulation using time lapse morphokinetics and preimplantation genetic testing in an IVF patient population. J Assist Reprod Genet. 2019;36(8):1561–9. https://doi.org/10.1007/s10815-019-01501-1.

    Article  PubMed  PubMed Central  Google Scholar 

  42. Nazem TG, Sekhon L, Lee JA, Overbey J, Pan S, Duke M, Briton-Jones C, Whitehouse M, Copperman AB, Stein DE. The correlation between morphology and implantation of euploid human blastocysts. Reprod BioMed Online. 2019;38(2):169–76. https://doi.org/10.1016/j.rbmo.2018.10.007.

    Article  PubMed  Google Scholar 

  43. Guzman L, Nuñez D, López R, Inoue N, Portella J, Vizcarra F, Noriega-Portella L, Noriega-Hoces L, Munné S. The number of biopsied trophectoderm cells may affect pregnancy outcomes. J Assist Reprod Genet. 2019;36(1):145–51. https://doi.org/10.1007/s10815-018-1331-1.

    Article  PubMed  Google Scholar 

  44. Casper RF, Yanushpolsky EH. Optimal endometrial preparation for frozen embryo transfer cycles: window of implantation and progesterone support. Fertil Steril. 2016;105(4):867–72. https://doi.org/10.1016/j.fertnstert.2016.01.006.

    Article  CAS  PubMed  Google Scholar 

  45. Lawrenz B, Coughlan C, Fatemi HM. Individualized luteal phase support. Curr Opin Obstet Gynecol. 2019;31(3):177–82. https://doi.org/10.1097/GCO.0000000000000530.

    Article  PubMed  Google Scholar 

  46. Lawrenz B, Coughlan C, Melado L, Fatemi HM. The ART of frozen embryo transfer: back to nature! Gynecol Endocrinol. 2020;36(6):479–83. https://doi.org/10.1080/09513590.2020.1740918.

    Article  PubMed  Google Scholar 

  47. Poletto KQ, Lobo MP, Giovanucci M, Approbato MS, Castro EC. Pregnancy rates from natural and artificial cycles of women submitted to frozen embryo transfers: a metanalysis. JBRA Assist Reprod. 2019;23(3):268–72. https://doi.org/10.5935/1518-0557.20190018.

    Article  PubMed  PubMed Central  Google Scholar 

  48. Kim MK, Park JK, Jeon Y, et al. Correlation between morphologic grading and euploidy rates of blastocysts, and clinical outcomes in in vitro fertilization preimplantation genetic screening. J Korean Med Sci. 2019;34(4):e27. Published 2019 Jan 10. https://doi.org/10.3346/jkms.2019.34.e27.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Maxwell SM, Grifo JA. Should every embryo undergo preimplantation genetic testing for aneuploidy? A review of the modern approach to in vitro fertilization. Best Pract Res Clin Obstet Gynaecol. 2018;53:38–47. https://doi.org/10.1016/j.bpobgyn.2018.07.005.

    Article  PubMed  Google Scholar 

  50. Zegers-Hochschild F, Adamson GD, Dyer S, Racowsky C, de Mouzon J, Sokol R, Rienzi L, Sunde A, Schmidt L, Cooke ID, Simpson JL, van der Poel S. The International Glossary on Infertility and Fertility Care, 2017. Fertil Steril. 2017;108(3):393–406. https://doi.org/10.1016/j.fertnstert.2017.06.005.

    Article  PubMed  Google Scholar 

Download references

Acknowledgements

We thank our biostatistician Ms. Gioia Altobelli for the statistical analysis and critical review of the manuscript.

Author information

Authors and Affiliations

  1. ART Fertility Clinics, Al Ain, Abu Dhabi, United Arab Emirates

    Andrea Abdala, Ibrahim Elkhatib, Aşina Bayram, Ana Arnanz, Ahmed El-Damen, Laura Melado, Barbara Lawrenz, Human M. Fatemi & Neelke De Munck

  2. Biomedicine and Biotechnology Department, University of Alcalá, Alcalá de Henares, Madrid, Spain

    Ana Arnanz

  3. Obstetrical Department, Women’s University Hospital Tuebingen, Tuebingen, Germany

    Barbara Lawrenz

Authors
  1. Andrea Abdala
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ibrahim Elkhatib
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Aşina Bayram
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ana Arnanz
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ahmed El-Damen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Laura Melado
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Barbara Lawrenz
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Human M. Fatemi
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Neelke De Munck
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

AAb, ND, HF: conception and design of the study; AAb, ND, IE, AAr, AD, AB, LM, BL, HF: oocyte retrieval; ND, IE, AAr, AD, AB, AAb: IVF and ICSI procedure and embryo evaluations; ND, IE, AAr, AD, AB, AAb: blastocyst biopsy, tubing and freezing; ND, IE, AB, AAr, LM, BL, HF: embryo transfers; AAb, GA: construction of the database and data interpretation; AAb: drafting the manuscript; ND, IE, AAr, AD, AB, LM, BL, HF: critical review and final approval of the manuscript.

Corresponding author

Correspondence to Andrea Abdala.

Ethics declarations

Conflict of interest

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.

Supplementary materials

ESM 1

(DOCX 42 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Abdala, A., Elkhatib, I., Bayram, A. et al. Day 5 vs day 6 single euploid blastocyst frozen embryo transfers: which variables do have an impact on the clinical pregnancy rates?. J Assist Reprod Genet 39, 379–388 (2022). https://doi.org/10.1007/s10815-021-02380-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-021-02380-1

Keywords

Search

Navigation