Comparative Embryo Development Outcomes following Extending Embryo Culture to Day 6: A Retrospective Cohort Study

Document Type : Original Article

Authors

1 Department of Obstetrics and Gynecology, University of Ottawa, Ottawa, Canada

2 Ottawa Fertility Centre, Ottawa, Canada

3 Ottawa Hospital Research Institute, Ottawa, Canada

4 Faculty of Medicine, University of Ottawa, Ottawa, Canada

Abstract

Background: Past studies have shown that culturing slow-growing embryos from day 5 to day 6 may increase vitrification yield. This study aims to evaluate if the proportion of embryos eligible for vitrification increases by growing embryos not vitrified by day 5 to day 6.
Materials and Methods: In this retrospective cohort study, a Canadian tertiary-care clinic-based cohort was identified between August 2019 and December 2020. In vitro fertilization (IVF) cycles involving autologous oocytes with at least one viable day 5 embryo were selected for inclusion. We compared embryo developmental outcomes of IVF cycles performed before and after an embryo cryopreservation policy change. Prior to March 2020, good-quality day 5 blastocysts of any stage were eligible for vitrification, and after that date, good-quality expanded blastocysts on either day 5 or day 6 were eligible. The primary outcome is the comparative proportion of embryos eligible for vitrification. The secondary outcome is to identify embryo, maternal and cycle factors that are predictive of day 6 vitrification.
Results: A total of 3,438 viable embryos across 679 consecutive IVF cycles were included in this study. After the policy change, we found similar mean proportions of blastocysts eligible for cryopreservation (46.9% per IVF cycle in group 2 vs. 44.4% in group 1, mean difference 0.025, 95% confidence interval -0.021 to 0.071, P=0.28). The mean number of cryopreserved embryos were significantly higher in group 2 (mean 2.2 vs. 1.7 embryos, P=0.007). Factors that predicated an embryo’s progression to day 6 included: younger age of egg provider, presence of an early blastocyst on day 5, and cycles involving surgically-retrieved sperm.
Conclusion: A cryopreservation policy change to include good-quality full and expanded day 6 blastocysts while avoiding to vitrify early blastocysts on day 5 yielded comparable proportions of embryos eligible for vitrification per IVF cycle.

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  1. Strauss J, Barbieri R. Yen & Jaffe's Reproductive Endocrinology. 8th ed. Philadelphia: Elsevier; 2018.
  2. Taylor TH, Patrick JL, Gitlin SA, Wilson JM, Crain JL, Griffin DK. Comparison of aneuploidy, pregnancy and live birth rates between day 5 and day 6 blastocysts. Reprod Biomed Online. 2014; 29(3): 305-310.
  3. 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(6): 1173-1181.
  4. Shapiro BS, Richter KS, Harris DC, Daneshmand ST. A comparison of day 5 and day 6 blastocyst transfers. Fertil Steril. 2001; 75(6): 1126-1130.
  5. Barrenetxea G, Lopez de Larruzea A, Ganzabal T, Jimenez R, Carbonero K, Mandiola M. Blastocyst culture after repeated failure of cleavage-stage embryo transfers: a comparison of day 5 and day 6 transfers. Fertil Steril. 2005; 83(1): 49-53.
  6. Shapiro BS, Daneshmand ST, Garner FC, Aguirre M, Ross R. Contrasting patterns in in vitro fertilization pregnancy rates among fresh autologous, fresh oocyte donor, and cryopreserved cycles with the use of day 5 or day 6 blastocysts may reflect differences in embryo-endometrium synchrony. Fertil Steril. 2008; 89(1): 20-26.
  7. Tubbing A, Shaw-Jackson C, Ameye L, Colin J, Rozenberg S, Autin C. Increased live births after day 5 versus day 6 transfers of vitrified-warmed blastocysts. J Assist Reprod Genet. 2018; 35(3): 417-424.
  8. Bourdon M, Pocate-Cheriet K, Finet de Bantel A, Grzegorczyk-Martin V, Amar Hoffet A, Arbo E, et al. Day 5 versus Day 6 blastocyst transfers: a systematic review and meta-analysis of clinical outcomes. Hum Reprod. 2019; 34(10): 1948-1964.
  9. Bilgory A, Kalma Y, Kopel R, Azem F. Transfer of day 6 frozen-thawed blastocysts on day 5 compared with day 6: catching up with the window of implantation-a retrospective study. Reprod Sci. 2021; 28(8): 2208-2215.
  10. 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): 1632-1639.
  11. Tannus S, Cohen Y, Henderson S, Al Ma'mari N, Shavit T, Son WY, et al. 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.
  12. El-Toukhy T, Wharf E, Walavalkar R, Singh A, Bolton V, Khalaf Y, et al. Delayed blastocyst development does not influence the outcome of frozen-thawed transfer cycles. BJOG. 2011; 118(13): 1551-1556.
  13. Elgindy E, Elsedeek MS. Day 5 expanded blastocysts transferred on same day have comparable outcome to those left for more extended culture and transferred on day 6. J Assist Reprod Genet. 2012; 29(10): 1111-1115.
  14. Wirleitner B, Schuff M, Stecher A, Murtinger M, Vanderzwalmen P. Pregnancy and birth outcomes following fresh or vitrified embryo transfer according to blastocyst morphology and expansion stage, and culturing strategy for delayed development. Hum Reprod. 2016; 31(8): 1685-1695.
  15. Gardner DK, Schoolcraft WB. In vitro culture of human blastocyst. In: Jansen, R, Mortimer D, editors, Towards reproductive certainty: infertility and genetics beyond. Carnforth: Parthenon Press; 377-388.
  16. Ivec M, Kovacic B, Vlaisavljevic V. Prediction of human blastocyst development from morulas with delayed and/or incomplete compaction. Fertil Steril. 2011; 96(6): 1473-1478.
  17. Kort JD, Lathi RB, Brookfield K, Baker VL, Zhao Q, Behr BR. Aneuploidy rates and blastocyst formation after biopsy of morulae and early blastocysts on day 5. J Assist Reprod Genet. 2015; 32(6): 925-930.
  18. Haas J, Meriano J, Bassil R, Barzilay E, Zilberberg E, Casper RF. Developmental potential of slow-developing embryos: day-5 morulae compared with day-5 cavitating morulae. Fertil Steril. 2019; 111(1): 105-111.
  19. Sallem A, Santulli P, Barraud-Lange V, Le Foll N, Ferreux L, Maignien C, et al. Extended culture of poor-quality supernumerary embryos improves ART outcomes. J Assist Reprod Genet. 2018; 35(2): 311-319.
  20. Tsai NC, Su YT, Lin YJ, Chiang HJ, Huang FJ, Kung FT, et al. Developmental potential of surplus morulas with delayed and/or incomplete compaction after freezing-thawing procedures. Reprod Biol Endocrinol. 2019; 17(1): 87.
  21. Wiemer KE, Dale B, Hu Y, Steuerwald N, Maxson WS, Hoffman DI. Blastocyst development in co-culture: development and morphological aspects. Hum Reprod. 1995; 10(12): 3226-3232.
  22. Boostanfar R, Jain JK, Slater CC, Tourgeman DE, Francis MM, Paulson RJ. The prognostic significance of day 3 embryo cleavage stage on subsequent blastocyst development in a sequential culture system. J Assist Reprod Genet. 2001; 18(10): 548-550.
  23. Shapiro BS, Harris DC, Richter KS. Predictive value of 72-hour blastomere cell number on blastocyst development and success of subsequent transfer based on the degree of blastocyst development. Fertil Steril. 2000; 73(3): 582-586.
  24. Hong YH, Kim HK, Nho EJ, Youm HW, Kim SK, Lee JR, et al. Predictors of blastocyst formation rate in elective day 5 transfer cycle. J Obstet Gynaecol. 2020; 40(6): 863-868.
  25. Scholtes MC, Zeilmaker GH. Blastocyst transfer in day-5 embryo transfer depends primarily on the number of oocytes retrieved and not on age. Fertil Steril. 1998; 69(1): 78-83.
  26. Yin H, Jiang H, He R, Wang C, Zhu J, Luan K. The effects of fertilization mode, embryo morphology at day 3, and female age on blastocyst formation and the clinical outcomes. Syst Biol Reprod Med. 2015; 61(1): 50-56.
  27. Wang C, Feng G, Zhang B, Shu J, Zhou H, Gan X, et al. Influence of the insemination method on the outcomes of elective blastocyst culture. Clin Exp Reprod Med. 2017; 44(2): 85-89.
  28. Miller JE, Smith TT. The effect of intracytoplasmic sperm injection and semen parameters on blastocyst development in vitro. Hum Reprod. 2001; 16(5): 918-924.
  29. Xu ZP, Sun HX, Hu YL, Zhang NY, Wang B. Effects of fertilization methods and sperm sources on the developmental capacity of surplus embryos. Zhonghua Nan Ke Xue. 2009; 15(10): 901-905.
  30. Westphal LM, Hinckley MD, Behr B, Milki AA. Effect of ICSI on subsequent blastocyst development and pregnancy rates. J Assist Reprod Genet. 2003; 20(3): 113-116.
  31. Van Landuyt L, De Vos A, Joris H, Verheyen G, Devroey P, Van Steirteghem A. Blastocyst formation in in vitro fertilization versus intracytoplasmic sperm injection cycles: influence of the fertilization procedure. Fertil Steril. 2005; 83(5): 1397-1403.
  32. Janny L, Menezo YJ. Evidence for a strong paternal effect on human preimplantation embryo development and blastocyst formation. Mol Reprod Dev. 1994; 38(1): 36-42.
  33. Kaing A, Kroener LL, Tassin R, Li M, Liu L, Buyalos R, et al. Earlier day of blastocyst development is predictive of embryonic euploidy across all ages: essential data for physician decision-making and counseling patients. J Assist Reprod Genet. 2018; 35(1): 119-125.