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Cryopreservation of Mammalian Oocytes: Slow Cooling and Vitrification as Successful Methods for Cryogenic Storage

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Cryopreservation and Freeze-Drying Protocols

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Abstract

Two basic 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. This method has also produced live births from cryopreserved human oocytes. The second method, which is described here, employs a high concentration of the cryoprotectant dimethyl sulfoxide plus a low concentration of polyethylene glycol. This is a vitrification method, which involves ultra-rapid 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 that have been vitrified using this technique have resulted in live births. Vitrification using other cryoprotectant mixtures is now a popular clinically accepted method for cryobanking of human oocytes.

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Acknowledgments

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.

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Authors and Affiliations

  1. Reproductive Medicine, Department of Obstetrics and Gynecology, Karolinska University Hospital, Stockholm, Sweden

    Victoria Keros

  2. ANOVA – Andrology, Sexual Medicine and Transmedicine, Department of Medicine (MedH), Karolinska Institutet, Stockholm, Sweden

    Victoria Keros

  3. Division of Surgery & Interventional Science, University College London Medical School, London, UK

    Barry J. Fuller

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  1. Victoria Keros
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  2. Barry J. Fuller
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Correspondence to Victoria Keros .

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Editors and Affiliations

  1. Unit for Reproductive Medicine—Clinic for Horses, Biostabilization Laboratory—Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, University of Veterinary Medicine Hannover, Hannover, Germany, University of Veterinary Medicine Hannover, Hannover, Germany

    Willem F. Wolkers

  2. Unit for Reproductive Medicine—Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany

    Harriëtte Oldenhof

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Keros, V., Fuller, B.J. (2021). Cryopreservation of Mammalian Oocytes: Slow Cooling and Vitrification as Successful Methods for Cryogenic Storage. In: Wolkers, W.F., Oldenhof, H. (eds) Cryopreservation and Freeze-Drying Protocols. Methods in Molecular Biology, vol 2180. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0783-1_20

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  • DOI: https://doi.org/10.1007/978-1-0716-0783-1_20

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