Abstract
A light modification on the mouse embryos CARD simple vitrification protocol makes it possible the oocyte freezing. This rapid and handy method for cryopreservation of mouse oocytes has many advantages and multiple applications like banking the female gamete for future use, controlling the IVF timing, and facilitating the transgenic production. In this chapter we described the protocol for cryopreserving oocytes by simple vitrification method and the in-vitro fertilization carried out using fresh or frozen sperm. With this method, nearly all of the cryopreserved oocytes were recovered, resulting in more than 90% morphologically normal. From those oocytes used for in vitro fertilization, 89% of the oocytes were fertilized.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Jaenisch R (1988) Transgenic animals. Science 240:1468–1473
Bedell MA, Largaespada DA, Jenkins NA, Copeland NG (1997) Mouse models of human disease. Part II: Recent progress and future directions. Genes Dev 11:11–43
Simpson EM, Linder CC, Sargent EE, Davisson MT, Mobraaten LE et al (1997) Genetic variation among 129 substrains and its importance for targeted mutagenesis in mice. Nat Genet 16:19–27
Brown SDM, Nolan PM (1998) Mouse mutagenesis – systematic studies of mammalian gene function. Hum Mol Genet 7(10):1627–1633
Hrabe de Angelis M, Balling R (1998) Large scale ENU screens in the mouse: genetics meets genomics. Mutat Res 400:25–32
Davisson M (2006) FIMRe: Federation of International Mouse Resources: global networking of resource centers. Mamm Genome 17:363–364
Donahue LR, Hrabe de Angelis M, Hagn M, Franklin C, Kent Lloyd KC, Magnuson T, McKerlie C, Nakagata N, Obata Y, Read S, Wurst W, Hörlein A, Davisson T (2012) Centralized mouse repositories. Mamm Genome 23(0):559–571
Parkening TA, Chang MC (1977) Effects of cooling rates and maturity of the animal on the recovery and fertilization of frozen-thawed rodent eggs. Biol Reprod 17:527–531
Whittingham DG (1977) Fertilization in vitro and development to term of unfertilized mouse oocytes previously stored at −196 degrees C. J Reprod Fertil 49:89–94
Nakagata N (1989) High survival rate of unfertilized mouse oocytes after vitrification. J Reprod Fertil 87(2):479–483
Nakagata N (1990) Cryopreservation of unfertilized mouse oocytes from inbred strains by ultrarapid freezing. Jikken Dobutsu 39(2):303–305
Nakagata N (1993) Production of normal young following transfer of mouse embryos obtained by in vitro fertilization between cryopreserved gametes. J Reprod Fertil 99:77–80
Shaw JM, Nakagata N (2002) Cryopreservation of transgenic mouse lines. Methods Mol Biol 180:207–228
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 57(6):675–680
Nakao K, Nakagata N, Katsuki M (1997) Simple and efficient procedure for cryopreservation of mouse embryos by simple vitrification. Exp Anim 46:231–234
Nakagata N, Takeo T, Fukumoto K, Kondo T, Haruguchi Y, Takeshita Y, Nakamuta Y, Matsunaga H, Tsuchiyama S, Ishizuka Y, Araki K (2013) Applications of cryopreserved unfertilized mouse oocytes for in vitro fertilization. Cryobiology 67(2):188–192
Nakagata N (1996) Use of cryopreservation techniques of embryos and spermatozoa for production of transgenic (Tg) mice and for maintenance of Tg mouse lines. Lab Anim Sci 46(2):236–238
Sakamoto W, Kaneko T, Nakagata N (2005) Use of frozen-thawed oocytes for efficient production of normal offspring from cryopreserved mouse spermatozoa showing low fertility. Comp Med 55(2):136–139
Nakagata N (2000) Cryopreservation of mouse spermatozoa. Mamm Genome 11:572–576
Nakagata N (2011) Cryopreservation of mouse spermatozoa and in vitro fertilization. Methods Mol Biol 693:57–73
Takeo T, Nakagata N (2011) Reduced glutathione enhances fertility of frozen/thawed C57BL/6 mouse sperm after exposure to methyl-beta-cyclodextrin. Biol Reprod 85(5):1066–1072
Takeo T, Tsutsumi A, Omaru T, Fukumoto K, Haruguchi Y, Kondo T, Nakamuta Y, Takeshita Y, Matsunaga H, Tsuchiyama S, Sakoh K, Nakao S, Yoshimoto H, Shimizu N, Nakagata N (2012) Establishment of a transport system for mouse epididymal sperm at refrigerated temperatures. Cryobiology 65(3):163–168
Quinn P, Kerin JF, Warnes GM (1985) Improved pregnancy rate in human in vitro fertilization with the use of a medium based on the composition of human tubal fluid. Fertil Steril 44:493–498
Whittingham DG (1974) Embryo banks in the future of developmental genetics. Genetics 78(1):395–402
Ho Y, Wigglesworth K, Eppig JJ, Schultz RM (1995) Preimplantation development of mouse embryos in KSOM: augmentation by amino acids and analysis of gene expression. Mol Reprod Dev 41:232–238
Nakagata N (1992) Embryo transfer through the wall of the Fallopian tube in mice. Exp Anim 41(3):387–388
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Nakagata, N., Sztein, J., Takeo, T. (2019). The CARD Method for Simple Vitrification of Mouse Oocytes: Advantages and Applications. In: Liu, C., Du, Y. (eds) Microinjection. Methods in Molecular Biology, vol 1874. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8831-0_13
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8831-0_13
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8830-3
Online ISBN: 978-1-4939-8831-0
eBook Packages: Springer Protocols