Skip to main content
SpringerLink
Log in

Hydroxypropyl cellulose supplementation in vitrification solutions: a prospective study with donor oocytes

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

Abstract

Purpose

Hydroxypropyl cellulose (HPC), a polysaccharide that forms a viscous gel under low temperatures, is a promising substitute of the blood-derived macromolecules traditionally used in cryopreservation solutions. The performance of a protein-free, fully synthetic set of vitrification and warming solutions was assessed in a matched pair analysis with donor oocytes.

Methods

A prospective study including 219 donor MII oocytes was carried out, comparing the laboratory outcomes of oocytes vitrified with HPC-based solutions and their fresh counterparts. The primary performance endpoint was the fertilization rate. Secondary parameters assessed were embryo quality on days 2 and 3.

Results

70/73 (95.9%) vitrified MII oocytes exhibited morphologic survival 2 h post-warming, with 49 (70.0%) presented normal fertilization, compared to 105 of 146 (71.9%) MII fresh oocytes. Similar embryo quality was observed in both groups. A total of 18 embryos implanted, out of 38 embryos transferred (47.3%), resulting in 13 newborns.

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

Similar content being viewed by others

References

  1. Rall WF, Fahy GM. Ice-free cryopreservation of mouse embryos at −196°C by vitrification. Nature. 1985;313:573–5.

    Article  CAS  PubMed  Google Scholar 

  2. Bakhach J. The cryopreservation of composite tissues: principles and recent advancement on cryopreservation of different type of tissues. Organogenesis. 2009;5(3):19–126.

    Google Scholar 

  3. Baudot A, Alger L, Boutron P. Glass-forming tendency in the system water-dimethyl sulfoxide. Cryobiology. 2000;40(2):151–8.

    Article  CAS  PubMed  Google Scholar 

  4. Benson EE. Cryopreservation theory; plant cryopreservation: a practical guide. Springer 2008

  5. Boutron P, Mehl P, Kaufmann A, Angibaud P. Glass-forming tendency and stability of the amorphous state in the aqueous solutions of linear polyalcohols with four carbons. I. Binary systems water-polyalcohol. Cryobiology. 1986;23(5):453–69.

    Article  CAS  PubMed  Google Scholar 

  6. Risco R, Elmoazzen H, Doughty M, He X, Toner M. Thermal performance of quartz capillaries for vitrification. Cryobiology. 2007;55(3):222–9.

    Article  CAS  PubMed  Google Scholar 

  7. Seki S, Mazur P. Ultra-rapid warming yields high survival of mouse oocytes cooled to −196 °C in dilutions of a standard vitrification solution. PLoS One. 2012;7:e36058.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Toner M, Carvalho EG, Karel M. Thermodynamics and kinetics of intracellular ice formation during freezing of biological cells. J App Phys. 1990;67:1582–93.

    Article  Google Scholar 

  9. Yavin S, Arav A. Measurement of essential physical properties of vitrification solutions. Theriogenology. 2007;67:81–9.

    Article  CAS  PubMed  Google Scholar 

  10. Wowk B. Thermodynamic aspects of vitrification. Cryobiology. 2010;60(1):11–22.

    Article  CAS  PubMed  Google Scholar 

  11. Cobo A, Serra V, Garrido N, Olmo I, Pellicer A, Remohí J. Obstetric and perinatal outcome of babies born from vitrified oocytes. Fertil Steril. 2014;102(4):1006–15.

    Article  PubMed  Google Scholar 

  12. Rienzi L, Ubaldi F, Iacobelli M, Ferrero S, Minasi MG, Martinez F, et al. Day 3 embryo transfer with combined evaluation at the pronuclear and cleavage stages compares favorably with day 5 blastocyst transfer. Hum Reprod. 2002;17:1852–5.

    Article  PubMed  Google Scholar 

  13. Solé M, Santaló J, Boada M, Clua E, Rodríguez I, Martínez 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;28:2087–92.

    Article  PubMed  Google Scholar 

  14. Practice Committee of the American Society for Reproductive Medicine, & Practice Committee of the Society for Assisted Reproductive Technology. Mature oocyte cryopreservation: a guideline. Fertil Steril. 2013;99(1):37–43.

    Article  Google Scholar 

  15. Ruffing NA, Steponkus PL, Pitt RE, Parks JE. Osmometric behavior, hydraulic conductivity, and incidence of intracellular ice formation in bovine oocytes at different developmental stages. Cryobiology. 1993;30:562–80.

    Article  CAS  PubMed  Google Scholar 

  16. Van den Abbeel E, Schneider U, Liu J, Agca Y, Critser JK, Van Steirteghem A. Osmotic responses and tolerance limits to changes in external osmolalities, and oolemma permeability characteristics, of human in vitro matured MII oocytes. Hum Reprod. 2007;22:1959–72.

    Article  PubMed  Google Scholar 

  17. Fahy GM. The relevance of cryoprotectant “toxicity” to cryobiology. Cryobiology. 1986;23:1–13.

    Article  CAS  PubMed  Google Scholar 

  18. Széll A, Shelton JN. Osmotic and cryoprotective effects of glycerolsucrose solutions on day-3 mouse embryos. J Reprod Fertil. 1987;80:309–16.

    Article  PubMed  Google Scholar 

  19. 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 

  20. Stachecki JJ, Garrisi J, Sabino S, Caetano JP, Wiemer K, Cohen J. A new safe, simple, and successful vitrification method for bovine and human blastocysts. Reprod Biomed Online. 2008;17:360–7.

    Article  PubMed  Google Scholar 

  21. Vanderzwalmen P, Connan D, Grobet L, Wirleitner B, Remy B, Vanderzwalmen S, et al. Lower intracellular concentration of cryoprotectants after vitrification than after slow freezing despite exposure to higher concentration of cryoprotectant solutions. Hum Reprod. 2013;28(8):2101–10.

    Article  CAS  PubMed  Google Scholar 

  22. Arav A. Cryopreservation of oocytes and embryos. 40th Anniversary Special Issue. Theriogenology. 2014;81:96–102.

    Article  CAS  PubMed  Google Scholar 

  23. Vajta G, Rienzi L, Cobo A, Yovich J. Embryo culture: can we perform better than nature? Reprod Biomed Online. 2010;20(4):453–69.

    Article  PubMed  Google Scholar 

  24. Shaw JM, Kuleshova LL, Macfarlane DR, Trounson AO. Vitrification properties of solutions of ethylene glycol in saline containing PVP, Ficoll, or dextran. Cryobiology. 1997;35:219–29.

    Article  CAS  PubMed  Google Scholar 

  25. Mori C, Yabuuchi A, Ezoe K, Murata N, Takayama Y, Okimura T, et al. Hydroxypropyl cellulose as an option for supplementation of cryoprotectant solutions for embryo vitrification in human assisted reproductive technologies. Reprod Biomed Online. 2015;30(6):613–21.

    Article  CAS  PubMed  Google Scholar 

  26. Inoue F. Hydroxypropyl cellulose as a macromolecular supplement for cryopreservation by vitrification of bovine ooytes and blastocysts and human oocytes. Fertil Steril. 2011;96(3):S212–3.

    Article  Google Scholar 

  27. Kuwayama M. Efficiency of non-protein solutions using hydroxypropyl cellulose on survival of bovine and human oocytes and embryos after vitrification. Fertil Steril. 2013;100(3):S174.

    Article  Google Scholar 

  28. Coello A, Campos P, Remohí J, Meseguer M, Cobo A. A combination of hydroxypropyl cellulose and trehalose as supplementation for vitrification of human oocytes: a retrospective cohort study. J Assist Reprod Genet. 2015;33(3):413–21.

    Article  Google Scholar 

  29. Clua E, Tur R, Coroleu B, Boada M, Barri PN, Veiga A. Analysis of factors associated with multiple pregnancy in an oocyte donation programme. Reprod Biomed Online. 2010;21:694–9.

    Article  PubMed  Google Scholar 

  30. Risco R, Hebles M, Saa AM, Vilches-Ferron AM, Sanchez-Martin P, Lucena E, et al. Safespeed technology—the myth of ultrahigh cooling rates: a close device and a serum-free media for the vitrification human oocytes/embryos with the highest recovery rates. Hum Reprod. 2013;28(S1):i180.

    Google Scholar 

  31. Kuwayama M, Vajta G, Kato O, Leibo SP. Highly efficient vitrification method for cryopreservation of human oocytes. Reprod Biomed Online. 2005;11:300–8.

    Article  PubMed  Google Scholar 

  32. Alpha SIRM. The Alpha consensus meeting on cryopreservation key performance indicators and benchmarks: proceedings of an expert meeting. Reprod Biomed Online. 2012;25(2):146.

    Article  Google Scholar 

  33. Cobo A, Kuwayama M, Pérez S, Ruiz A, Pellicer A, Remohí J. Comparison of concomitant outcome achieved with fresh and cryopreserved donor oocytes vitrified by the Cryotop method. Fertil Steril. 2008;89:1657–64.

    Article  PubMed  Google Scholar 

  34. Maggiulli R, Scarica C, Dovere L, Ievoli E, Ubaldi FM, Rienzi L. Efficiency of human oocyte and embryo vitrification. Curr Trends Clin Embriol. 2015;1:2–11.

    Google Scholar 

  35. Rienzi L, Romano S, Albricci L, Maggiulli R, Capalbo A, Baroni E, et al. Embryo development of fresh ‘versus’ vitrified metaphase II oocytes after ICSI: a prospective randomized sibling-oocyte study. Hum Reprod. 2010;25(1):66–73.

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

We would like to give special thanks to all the staff of the clinic that participated in the study. We would also like to express our gratitude for the excellent peer-review process the manuscript was subjected to.

Author information

Authors and Affiliations

  1. Clínica Ginemed, C/ Farmacéutico Murillo Herrera n 3-5, 41010, Seville, Spain

    Miguel Gallardo, María Hebles, Beatriz Migueles, Mónica Dorado, Laura Aguilera, Mercedes González, Paloma Piqueras, Lorena Montero, Pascual Sánchez-Martín & Fernando Sánchez-Martín

  2. Seville Engineering School, University of Seville, Camino de los Descubrimientos s/n, 41092, Seville, Spain

    Miguel Gallardo, Alejandro Lucas & Ramón Risco

  3. National Accelerators Centre, Calle Thomas Alva Edison, 7, 41092, Seville, Spain

    Ramón Risco

Authors
  1. Miguel Gallardo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. María Hebles
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Beatriz Migueles
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mónica Dorado
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Laura Aguilera
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mercedes González
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Paloma Piqueras
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Alejandro Lucas
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Lorena Montero
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Pascual Sánchez-Martín
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Fernando Sánchez-Martín
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Ramón Risco
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ramón Risco.

Ethics declarations

The study was conducted as described in the protocol, which was developed in accordance to the principles of Helsinki and the national Policy of Good Clinical Practice (ISO 14155:2011), and was approved by the internal review board of the institutional research committee. All recipients were informed thoroughly about the protocol of the study and signed an informed consent form.

Additional information

Capsule The satisfactory results of this direct comparison with fresh controls strongly support the effectiveness of HPC supplementation of vitrification solutions, confirming its role as a substitute for protein supplementation.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Gallardo, M., Hebles, M., Migueles, B. et al. Hydroxypropyl cellulose supplementation in vitrification solutions: a prospective study with donor oocytes. J Assist Reprod Genet 34, 417–422 (2017). https://doi.org/10.1007/s10815-016-0841-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-016-0841-y

Keywords

Search

Navigation