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Chapter 8 Human Oocytes Slow-Rate Freezing: Methodology

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Cryopreservation of Mammalian Gametes and Embryos

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1568))

Abstract

Cryopreservation of human oocytes is an important technique for the treatment of human infertility, as it deals successfully with legal, ethical, and moral issues related to embryo cryopreservation (Coticchio et al., Human Fertil (Camb) 4:152–157, 2001; Tucker et al., Eur J Obstet Gynecol Reprod Biol 113: 524–527, 2004) and maintains reproductive potential in women with diseases or conditions that may compromise reproductive capacity. Here, we describe an oocyte cryopreservation technique involving slow freezing–rapid thawing with differential sucrose concentration (0.2 M in the freezing stage—0.3 M in thawing stage) (Bianchi et al., Reprod Biomed Online. 14:64–71, 2007), describing the technique in detail, from the preparation of the solutions through to the performance of the technique and, finally, to a number of helpful hints for optimum results.

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References

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

    Article  Google Scholar 

  2. Coticchio G, Garetti S, Bonu MA, Borini A (2001) Cryopreservation of human oocytes. Hum Fertil (Camb) 4:152–157

    Article  CAS  Google Scholar 

  3. Tucker M, Morton P, Liebermann J (2004) Human oocyte cryopreservation: a valid alternative to embryo cryopreservation? Eur J Obstet Gynecol Reprod Biol 113(Suppl 1):524–527

    Google Scholar 

  4. Mazur P (1963) Kinetics of water loss from cells at subzero temperatures and the likelihood of intracellular freezing. J Gen Physiol 47:347–369

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Kleinhans FW, Mazur P (2007) Comparison of actual vs. synthesized ternary phase diagrams for solutes of cryobiological interest. Cryobiology 54:212–2225

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Chen C (1986) Pregnancy after human oocyte cryopreservation. Lancet 1:884–886

    Article  CAS  PubMed  Google Scholar 

  7. Al-Hasani DK, van der Ven H, Reinecke A et al (1987) Cryopreservation of human oocytes. Hum Reprod 2:695–700

    Article  CAS  PubMed  Google Scholar 

  8. Bianchi V, Coticchio G, Distratis V, Di Giusto N, Flamigni C, Borini A (2007) Differential sucrose concentration during dehydration (0.2 mol/L) and rehydration (0.3 mol/L) increases the implantation rate of frozen human oocytes. Reprod Biomed Online 14:64–71

    Article  CAS  PubMed  Google Scholar 

  9. Fabbri R, Porcu E, Marsella T, Rocchetta G, Venturoli S, Flamigni C (2001) Human oocytes cryopreservation: new prodpectives regarding oocytes survival. Hum Reprod 16:411–416

    Article  CAS  PubMed  Google Scholar 

  10. Stacheki JJ, Willadsen SM (2000) Cryopreservation of mouse oocytes using a medium with low sodium content: effect of plunge temperature. Cryobiology 40:4–12

    Article  Google Scholar 

  11. Boldt J, Tidswell N, Sayers A, Kilani R, Cline D (2006) Human oocytes cryopreservation: 5-year experience with a sodium-depleted slow freezing method. Reprod Biomed Online 13:96–100

    Article  CAS  PubMed  Google Scholar 

  12. Elliot K, Whelan J (eds) (1977) CIBA foundation symposium 52. The freezing of mammalian embryos. Elsevier, Amsterdam

    Google Scholar 

  13. MJ A–S, Farrant J (eds) (1980) Low temperature preservation in medicine and biology. Pitman Press, Bath UK

    Google Scholar 

  14. Lassalle B, Testart J, Renard JP (1985) Human embryo features that influence the success of cryopreservation with the use of 1,2 propanediol. Fertil Steril 44:645–651

    Article  CAS  PubMed  Google Scholar 

  15. Fabbri R, Porcu E, Marsella T et al (2001) Human oocytes cryopreservation: new perspectives regarding oocytes survival. Hum Reprod 16:411–416

    Article  CAS  PubMed  Google Scholar 

  16. Ashwood-Smith MJ (1987) Mechanisms of cryoprotectant action. Symp Soc Exp Biol. 41:395–406

    CAS  PubMed  Google Scholar 

  17. Jain JK, Paulson RJ (2006 Oct) Oocyte cryopreservation. Fertil Steril 86(4 Suppl):1037–1046

    Article  CAS  PubMed  Google Scholar 

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Correspondence to A. Borini .

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Zacà, C., Borini, A. (2017). Chapter 8 Human Oocytes Slow-Rate Freezing: Methodology. In: Nagy, Z., Varghese, A., Agarwal, A. (eds) Cryopreservation of Mammalian Gametes and Embryos. Methods in Molecular Biology, vol 1568. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6828-2_8

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  • DOI: https://doi.org/10.1007/978-1-4939-6828-2_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6826-8

  • Online ISBN: 978-1-4939-6828-2

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