Journal of Assisted Reproduction and Genetics

, Volume 33, Issue 9, pp 1247–1252 | Cite as

Closed vitrification of human oocytes and blastocysts: outcomes from a series of clinical cases

  • Debra A. GookEmail author
  • Boon Choo
  • Harold Bourne
  • Kelly Lewis
  • David H. Edgar
Gamete Biology



High survival rates and clinical outcomes similar to those from fresh oocytes and blastocysts have been observed with open oocyte vitrification systems. It has been suggested that the extremely fast cooling rates that are only achieved with open systems are necessary for human oocyte and blastocyst vitrification. However, there is a potential risk of introducing contamination with open systems. The aim of this study was to assess whether similar survival and subsequent implantation rates could be achieved using a closed vitrification system for human oocytes and blastocysts.


Initially, donated immature oocytes that were matured in vitro were vitrified using the cryoprotectants ethylene glycol (EG) + dimethyl sulphoxide (DMSO) + sucrose and either a closed system (Rapid-i®) or an open system (Cryolock). The closed system was subsequently introduced clinically for mature oocyte cryopreservation cases and blastocyst vitrification.


Using in vitro matured oocytes, a similar survival was achieved with the open system of 92.4 % (73/79) and with the closed system of 89.7 % (35/39). For clinical oocyte closed vitrification, high survival rate of 90.5 % (374/413) and an implantation rate of 32.7 % (18/55) from the transfer of day 2 embryos was achieved, which is similar to fresh day 2 embryo transfers. Blastocysts have also been successfully cryopreserved using the Rapid-i closed vitrification system with 94 % of blastocysts having an estimated ≥75 % of cells intact and a similar implantation rate (31.5 %) to fresh single blastocyst transfers.


Closed vitrification can achieve high survival and similar implantation rates to fresh for both oocytes and blastocysts.


Closed vitrification Human Oocyte Blastocyst Implantation rate Births 


  1. 1.
    Li Z, Wang YA, Ledger W, Edgar DH, Sullivan EA. Clinical outcomes following cryopreservation of blastocysts by vitrification or slow freezing: a population-based cohort study. Hum Reprod. 2014;29(12):2794–801.CrossRefPubMedGoogle Scholar
  2. 2.
    Edgar DH, Gook DA. A critical appraisal of cryopreservation (slow cooling versus vitrification) of human oocytes and embryos. Hum Reprod Update. 2012;18:536–54.CrossRefPubMedGoogle Scholar
  3. 3.
    Cobo A, de los Santos MJ, Castello D, Gamiz P, Campos P, Remohi J. Outcomes of vitrified early cleavage-stage and blastocyst-stage embryos in a cryopreservation program: evaluation of 3,150 warming cycles. Fertil Steril. 2012;98(5):1138–46. e1.CrossRefPubMedGoogle Scholar
  4. 4.
    Cobo A, Meseguer M, Remohi J, Pellicer A. Use of cryo-banked oocytes in an ovum donation programme: a prospective, randomized, controlled, clinical trial. Hum Reprod. 2010;25(9):2239–46.CrossRefPubMedGoogle Scholar
  5. 5.
    Cobo A, Remohi J, Chang CC, Nagy ZP. Oocyte cryopreservation for donor egg banking. Reprod Biomed Online. 2011;23(3):341–6.CrossRefPubMedGoogle Scholar
  6. 6.
    Liebermann J. Vitrification of human blastocysts: an update. Reprod Biomed Online. 2009;19 Suppl 4:4328.PubMedGoogle Scholar
  7. 7.
    Devine K, Connell MT, Richter KS, Ramirez CI, Levens ED, DeCherney AH, et al. Single vitrified blastocyst transfer maximizes liveborn children per embryo while minimizing preterm birth. Fertil Steril. 2015;103(6):1454–60. e1.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Bielanski A, Nadin-Davis S, Sapp T, Lutze-Wallace C. Viral contamination of embryos cryopreserved in liquid nitrogen. Cryobiology. 2000;40:110–6.CrossRefPubMedGoogle Scholar
  9. 9.
    Tedder RS, Zuckerman MA, Goldstone AH, Hawkins AE, Fielding A, Briggs EM, et al. Hepatitis B transmission from contaminated cryopreservation tank. Lancet. 1995;346(8968):137–40.CrossRefPubMedGoogle Scholar
  10. 10.
    Bielanski A, Vajta G. Risk of contamination of germplasm during cryopreservation and cryobanking in IVF units. Hum Reprod. 2009;24:2457–67.CrossRefPubMedGoogle Scholar
  11. 11.
    Cobo A, Romero JL, Perez S, de Los Santos MJ, Meseguer M, Remohi J. Storage of human oocytes in the vapor phase of nitrogen. Fertil Steril. 2010;94(5):1903–7.CrossRefPubMedGoogle Scholar
  12. 12.
    Parmegiani L, Accorsi A, Cognigni GE, Bernardi S, Troilo E, Filicori M. Sterilization of liquid nitrogen with ultraviolet irradiation for safe vitrification of human oocytes or embryos. Fertil Steril. 2010;94(4):1525–8.CrossRefPubMedGoogle Scholar
  13. 13.
    Parmegiani L, Accorsi A, Bernardi S, Arnone A, Cognigni GE, Filicori M. A reliable procedure for decontamination before thawing of human specimens cryostored in liquid nitrogen: three washes with sterile liquid nitrogen (SLN2). Fertil Steril. 2012;98(4):870–5.CrossRefPubMedGoogle Scholar
  14. 14.
    Vajta G, Rienzi L, Ubaldi FM. Open versus closed systems for vitrification of human oocytes and embryos. Reprod Biomed Online. 2015;30(4):325–33.CrossRefPubMedGoogle Scholar
  15. 15.
    Seki S, Mazur P. Effect of warming rate on the survival of vitrified mouse oocytes and on the recrystallization of intracellular ice. Biol Reprod. 2008;79(4):727–37.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Seki S, Mazur P. The dominance of warming rate over cooling rate in the survival of mouse oocytes subjected to a vitrification procedure. Cryobiology. 2009;59(1):75–82.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Vanderzwalmen P, Zech N, Prapas Y, Panagiotidis Y, Papatheodorou A, Lejeune B, et al. Closed carrier device: a reality to vitrify oocytes and embryos in aseptic conditions. Gynecol Obstet Fertil. 2010;38(9):541–6.CrossRefPubMedGoogle Scholar
  18. 18.
    Papatheodorou A, Vanderzwalmen P, Panagiotidis Y, Prapas N, Zikopoulos K, Georgiou I, et al. Open versus closed oocyte vitrification system: a prospective randomized sibling-oocyte study. Reprod Biomed Online. 2013;26(6):595–602.CrossRefPubMedGoogle Scholar
  19. 19.
    Chen Y, Zheng X, Yan J, Qiao J, Liu P. Neonatal outcomes after the transfer of vitrified blastocysts: closed versus open vitrification system. Reprod Biol Endocrinol. 2013;11:107.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Iwahata H, Hashimoto S, Inoue M, Inoue T, Ito K, Nakaoka Y, et al. Neonatal outcomes after the implantation of human embryos vitrified using a closed-system device. J Assist Reprod Genet. 2015;32(4):521–6.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Panagiotidis Y, Vanderzwalmen P, Prapas Y, Kasapi E, Goudakou M, Papatheodorou A, et al. Open versus closed vitrification of blastocysts from an oocyte-donation programme: a prospective randomized study. Reprod Biomed Online. 2013;26(5):470–6.CrossRefPubMedGoogle Scholar
  22. 22.
    Desai NN, Goldberg JM, Austin C, Falcone T. The new Rapid-i carrier is an effective system for human embryo vitrification at both the blastocyst and cleavage stage. Reprod Biol Endocrinol. 2013;11:41.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Hashimoto S, Amo A, Hama S, Ohsumi K, Nakaoka Y, Morimoto Y. A closed system supports the developmental competence of human embryos after vitrification : closed vitrification of human embryos. J Assist Reprod Genet. 2013;30(3):371–6.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Paffoni A, Guarneri C, Ferrari S, Restelli L, Nicolosi AE, Scarduelli C, et al. Effects of two vitrification protocols on the developmental potential of human mature oocytes. Reprod Biomed Online. 2011;22:292–8.CrossRefPubMedGoogle Scholar
  25. 25.
    Bourne H, Richings N, Harari O, Watkins W, Speirs AL, Johnston WIH, et al. The use of intracytoplasmic sperm injection for the treatment of severe and extreme male infertility. Reprod Fertil Dev. 1995;7:237–45.CrossRefPubMedGoogle Scholar
  26. 26.
    Lawler C, Baker HW, Edgar DH. Relationships between timing of syngamy, female age and implantation potential in human in vitro-fertilised oocytes. Reprod Fertil Dev. 2007;19(3):482–7.CrossRefPubMedGoogle Scholar
  27. 27.
    Edgar DH, Bourne H, Speirs AL, McBain JC. A quantitative analysis of the impact of cryopreservation on the implantation potential of human early cleavage stage embryos. Hum Reprod. 2000;15:175–9.CrossRefPubMedGoogle Scholar
  28. 28.
    Edgar DH, Karani J, Gook DA. Increasing dehydration of human cleavage-stage embryos prior to slow cooling significantly increases cryosurvival. Reprod Biomed Online. 2009;19(4):521–5.CrossRefPubMedGoogle Scholar
  29. 29.
    Nagy ZP, Chang CC, Shapiro DB, Bernal DP, Elsner CW, Mitchell-Leef D, et al. Clinical evaluation of the efficiency of an oocyte donation program using egg cryo-banking. Fertil Steril. 2009;92(2):520–6.CrossRefPubMedGoogle Scholar
  30. 30.
    Ubaldi F, Anniballo R, Romano S, Baroni E, Albricci L, Colamaria S, et al. Cumulative ongoing pregnancy rate achieved with oocyte vitrification and cleavage stage transfer without embryo selection in a standard infertility program. Hum Reprod. 2010;25(5):1199–205.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Garcia JI, Noriega-Portella L, Noriega-Hoces L. Efficacy of oocyte vitrification combined with blastocyst stage transfer in an egg donation program. Hum Reprod. 2011;26:782–90.CrossRefPubMedGoogle Scholar
  32. 32.
    Trokoudes KM, Pavlides C, Zhang X. Comparison outcome of fresh and vitrified donor oocytes in an egg-sharing donation program. Fertil Steril. 2011;95(6):1996–2000.CrossRefPubMedGoogle Scholar
  33. 33.
    Rienzi L, Cobo A, Paffoni A, Scarduelli C, Capalbo A, Vajta G, et al. Consistent and predictable delivery rates after oocyte vitrification: an observational longitudinal cohort multicentric study. Hum Reprod. 2012;27:1606–12.CrossRefPubMedGoogle Scholar
  34. 34.
    Sole M, Santalo J, Boada M, Clua E, Rodriguez I, Martinez F, et al. How does vitrification affect oocyte viability in oocyte donation cycles? A prospective study to compare outcomes achieved with fresh versus vitrified sibling oocytes. Hum Reprod. 2013;28(8):2087–92.CrossRefPubMedGoogle Scholar
  35. 35.
    Stoop D, De Munck N, Jansen E, Platteau P, Van den Abbeel E, Verheyen G, et al. Clinical validation of a closed vitrification system in an oocyte-donation programme. Reprod Biomed Online. 2012;24:180–5.CrossRefPubMedGoogle Scholar
  36. 36.
    De Munck N, Verheyen G, Van Landuyt L, Stoop D, Van de Velde H. Survival and post-warming in vitro competence of human oocytes after high security closed system vitrification. J Assist Reprod Genet. 2013;30(3):361–9.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Seki S, Mazur P. Ultra-rapid warming yields high survival of mouse oocytes cooled to −196 degrees C in dilutions of a standard vitrification solution. PLoS ONE. 2012;7(4):e36058.CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Gook DA, Edgar DH. Implantation rates of embryos generated from slow cooled human oocytes from young women are comparable to those of fresh and frozen embryos from the same age group. J Assist Reprod Genet. 2011;28(12):1171–6.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Debra A. Gook
    • 1
    • 2
    Email author
  • Boon Choo
    • 1
  • Harold Bourne
    • 1
  • Kelly Lewis
    • 1
  • David H. Edgar
    • 1
    • 2
  1. 1.Reproductive Services and Melbourne IVFRoyal Women’s HospitalParkvilleAustralia
  2. 2.Department of Obstetrics and GynaecologyUniversity of MelbourneParkvilleAustralia

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