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Cryopreservation of Mammalian Oocytes

  • Victoria KerosEmail author
  • Barry J. Fuller
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1257)

Abstract

Two methods for the laboratory-focused cryopreservation of mammalian oocytes are described, based on work with murine oocytes. One method uses a relatively low concentration of the cryoprotectant propanediol plus sucrose and requires controlled rate cooling equipment to achieve a slow cooling rate. Such a method has also produced live births from cryopreserved human oocytes. The second method described employs a high concentration of the cryoprotectant dimethyl sulfoxide plus a low concentration of polyethylene glycol. This is a vitrification method which involves ultrarapid cooling by plunging standard straws into liquid nitrogen vapor, hence avoiding the need for specialized equipment, but requires technical ability to manipulate the oocytes quickly in the highly concentrated cryoprotectant solutions. Murine oocytes vitrified using this technique has resulted in live births.

Key words

Oocyte Slow-cooling Vitrification Murine Human 

Notes

Acknowledgement

We would like to gratefully acknowledge the significant role played by Dr Sharon Paynter, who as a collaborator over many years developed the practical aspects of the two methods described in this chapter.

References

  1. 1.
    Whittingham DG (1977) Fertilization in vitro and development to term of unfertilized mouse oocytes previously stored at –196. J Reprod Fertil 49:89–94CrossRefGoogle Scholar
  2. 2.
    Al-Hasani A, Kirsch J, Diedrich K, Blanke S, van der Ven H, Krebs D (1989) Successful embryo transfer of cryopreserved and in vitro fertilised rabbit oocytes. Hum Reprod 4:77–79Google Scholar
  3. 3.
    Fuku E, Kojima T, Shoiya Y, Marcus GJ, Downey BR (1992) In vitro fertilization and development of frozen-thawed bovine oocytes. Cryobiology 29:485–492CrossRefGoogle Scholar
  4. 4.
    MacLellan LJ, Carnevale EM, da Silva MAC, Scoggin CF, Bruemmer JE, Squires EL (2002) Pregnancies from vitrified equine oocytes collected from super-stimulated and non-stimulated mares. Theriogenology 58:911–919CrossRefGoogle Scholar
  5. 5.
    Pope CE, Gómez MC, Kagawa N, Kuwayama M, Leibo SP, Dresser BL (2012) In vivo survival of domestic cat oocytes after vitrification, intracytoplasmic sperm injection and embryo transfer. Theriogenology 77:531–538CrossRefGoogle Scholar
  6. 6.
    Chen C (1986) Pregnancy after human oocyte cryopreservation. Lancet 1:884–886CrossRefGoogle Scholar
  7. 7.
    Bleil JD, Wassarman PM (1980) Structure and function of the zona pellucida: identification and characterization of the proteins of the mouse oocyte’s zona pellucida. Dev Biol 76:185–202CrossRefGoogle Scholar
  8. 8.
    Carroll J, Depypere H, Matthews CD (1990) Freeze-thaw induced changes of the zona pellucida explains decreased rates of fertilisation in frozen-thawed mouse oocytes. J Reprod Fertil 90:547–553CrossRefGoogle Scholar
  9. 9.
    Larman MG, Sheehan CB, Gardner DK (2006) Calcium-free vitrification reduces cryoprotectant-induced zona pellucida hardening and increases fertilization rates in mouse oocytes. Reproduction 131:53–61CrossRefGoogle Scholar
  10. 10.
    Bianchi V, Coticchio G, Fava L, Flamigni C, Borini A (2005) Meiotic spindle imaging in human oocytes frozen with a slow freezing procedure involving high sucrose concentration. Hum Reprod 20:1078–1083CrossRefGoogle Scholar
  11. 11.
    Stachecki JJ, Cohen J (2004) An overview of oocyte cryopreservation. Reprod Biomed Online 9:152–163CrossRefGoogle Scholar
  12. 12.
    Rienzi L, Martinez F, Ubaldi F, Minasi MG, Iacobelli M, Tesarik J, Greco E (2004) Polscope analysis of meiotic spindle changes in living metaphase II human oocytes during the freezing and thawing procedures. Hum Reprod 19:655–659CrossRefGoogle Scholar
  13. 13.
    Brower PT, Schultz RM (1982) Intercellular communication between granulosa cells and mouse oocytes: existence and possible nutritional role during oocyte growth. Dev Biol 90:144–153CrossRefGoogle Scholar
  14. 14.
    Ruppert-Lingham CJ, Paynter SJ, Godfrey J, Fuller BJ, Shaw RW (2003) Developmental potential of murine germinal vesicle stage cumulus-oocyte complexes following exposure to dimethylsulphoxide or cryopreservation: loss of membrane integrity of cumulus cells after thawing. Hum Reprod 18:392–398CrossRefGoogle Scholar
  15. 15.
    Keros V, Xella S, Hultenby K, Pettersson K, Sheikhi M, Volpe A, Hreinsson J, Hovatta O (2009) Vitrification versus controlled-rate freezing in cryopreservation of human ovarian tissue. Hum Reprod 24:1670–1683CrossRefGoogle Scholar
  16. 16.
    Isachenko V, Soler C, Isachenko E, Perez-Sanchez F, Grischenko V (1998) Vitrification of immature porcine oocytes: effects of lipid droplets, temperature, cytoskeleton, and addition and removal of cryoprotectant. Cryobiology 36:250–253CrossRefGoogle Scholar
  17. 17.
    Porcu E, Fabbri R, Seracchioli R, Ciotti PM, Magrini O, Flamigni C (1997) Birth of a healthy female after intracytoplasmic sperm injection of cryopreserved human oocytes. Fertil Steril 68:724–726CrossRefGoogle Scholar
  18. 18.
    Bianchi V, Lappi M, Bonu MA, Borini A (2012) Oocyte slow freezing using a 0.2–0.3 M sucrose concentration protocol: is it really the time to trash the cryopreservation machine? Fertil Steril 97:1101–1107CrossRefGoogle Scholar
  19. 19.
    Kuleshova L, Gianaroli L, Magli C, Ferraretti A, Trounson A (1999) Birth following vitrification of a small number of human oocytes. Hum Reprod 14:3077–3079CrossRefGoogle Scholar
  20. 20.
    Yoon TK, Chung HM, Lim JM, Han SY, Ko JJ, Cha KY (2000) Pregnancy and delivery of healthy infants developed from vitrified oocytes in a stimulated in vitro fertilization-embryo transfer program. Fertil Steril 74:180–181CrossRefGoogle Scholar
  21. 21.
    Noyes N, Porcu E, Borini A (2009) Over 900 oocyte cryopreservation babies born with no apparent increase in congenital anomalies. Reprod Biomed Online 18:769–776CrossRefGoogle Scholar
  22. 22.
    Kuwayama M, Vajta G, Kato O, Leibo SP (2005) Highly efficient vitrification method for cryopreservation of human oocytes. Reprod Biomed Online 11:300–308CrossRefGoogle Scholar
  23. 23.
    Kuwayama M (2007) Highly efficient vitrification for cryopreservation of human oocytes and embryos: the Cryotop method. Theriogenology 67:73–80CrossRefGoogle Scholar
  24. 24.
    Cao YX, Xing Q, Li L, Cong L, Zhang ZG, Wei ZL, Zhou P (2009) Comparison of survival and embryonic development in human oocytes cryopreserved by slow-freezing and vitrification. Fertil Steril 92:1306–1311CrossRefGoogle Scholar
  25. 25.
    Fadini R, Brambillasca F, Renzini MM, Merola M, Comi R, De Ponti E, Dal Canto MB (2009) Human oocyte cryopreservation: comparison between slow and ultrarapid methods. Reprod Biomed Online 19:171–180CrossRefGoogle Scholar
  26. 26.
    Edgar DH, Gook DA (2012) A critical appraisal of cryopreservation (slow cooling versus vitrification) of human oocytes and embryos. Hum Reprod Update 18:536–554CrossRefGoogle Scholar
  27. 27.
    Noyes N, Knopman J, Labella P, McCaffrey C, Clark-Williams M, Grifo J (2010) Oocyte cryopreservation outcomes including pre-cryopreservation and post-thaw meiotic spindle evaluation following slow cooling and vitrification of human oocytes. Fertil Steril 94:2078–2082CrossRefGoogle Scholar
  28. 28.
    Cobo A, Diaz C (2012) Clinical application of oocyte vitrification: a systematic review and meta-analysis of randomized controlled trials. Fertil Steril 96:277–285CrossRefGoogle Scholar
  29. 29.
    Paynter SJ (2000) Current status of the cryopreservation of human unfertilised oocytes. Hum Reprod Update 6:449–456CrossRefGoogle Scholar
  30. 30.
    Boldt J (2011) Current results with slow freezing and vitrification of the human oocyte. Reprod Biomed Online 23:314–322CrossRefGoogle Scholar
  31. 31.
    Kim TJ, Hong SW (2011) Successful live birth from vitrified oocytes after 5 years of cryopreservation. J Assist Reprod Genet 28:73–76CrossRefGoogle Scholar
  32. 32.
    Fabbri R, Porcu E, Marsella T, Rocchetta G, Venturoli S, Flamigni C (2001) Human oocyte cryopreservation: new perspectives regarding oocyte survival. Hum Reprod 16:411–416CrossRefGoogle Scholar
  33. 33.
    Paynter SJ, Borini A, Bianchi V, De Santis L, Flamigni C, Coticchio G (2005) Volume changes of mature human oocytes on exposure to cryoprotectant solutions used in slow cooling procedures. Hum Reprod 20:1194–1199CrossRefGoogle Scholar
  34. 34.
    Stachecki JJ, Cohen J, Willadsen SM (1998) Cryopreservation of unfertilized mouse oocytes: the effect of replacing sodium with choline in the freezing medium. Cryobiology 37:346–354CrossRefGoogle Scholar
  35. 35.
    Quintans CJ, Donaldson MJ, Bertolino MV, Pasqualini RS (2002) Birth of two babies using oocytes that were cryopreserved in a choline-based freezing medium. Hum Reprod 17:3149–3152CrossRefGoogle Scholar
  36. 36.
    Boldt J, Cline D, McLaughlin D (2003) Human oocyte cryopreservation as an adjunct to IVF-embryo transfer cycles. Hum Reprod 18:1250–1255CrossRefGoogle Scholar
  37. 37.
    Fahy GM, Wowk B, Wu J, Paynter SJ (2004) Improved vitrification solutions based on the predictability of vitrification solution toxicity. Cryobiology 48:22–35CrossRefGoogle Scholar
  38. 38.
    O’Neil L, Paynter SJ, Fuller BJ, Shaw RW (1997) Vitrification of mature mouse oocytes: improved results following addition of polyethylene glycol to a dimethyl sulfoxide solution. Cryobiology 34:295–301CrossRefGoogle Scholar
  39. 39.
    O’Neil L, Paynter SJ, Fuller BJ, Shaw RW (1999) Birth of live young from mouse oocytes vitrified in 6 M dimethyl sulfoxide supplemented with 1 mg/ml polyethylene glycol. Cryobiology 39:284Google Scholar
  40. 40.
    Kohaya N, Fujiwara K, Ito J, Kashiwazaki N (2011) High developmental rates of mouse oocytes cryopreserved by an optimized vitrification protocol: the effects of cryoprotectants, calcium and cumulus cells. J Reprod Dev 57:675–680CrossRefGoogle Scholar
  41. 41.
    Watanabe H, Kohaya N, Kamoshita M, Fujiwara K, Matsumura K, Hyon SH, Ito J, Kashiwazaki N (2013) Efficient production of live offspring from mouse oocytes vitrified with a novel cryoprotective agent, carboxylated ε-poly-l-lysine. PLoS One 8:e83613CrossRefGoogle Scholar
  42. 42.
    De Munck N, Verheyen G, Van Landuyt L, Stoop D, Van de Velde H (2013) Survival and post-warming in vitro competence of human oocytes after high security closed system vitrification. J Assist Reprod Genet 30:361–369CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Centre for Andrology and Sexual Medicine, Department of MedicineKarolinska InstitutetStockholmStockholm
  2. 2.Reproductive MedicineKarolinska University HospitalStockholmSweden
  3. 3.University College London Medical SchoolLondonUK

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