Skip to main content
SpringerLink
Log in

Cryopreservation

  • Protocol
  • First Online:
Advanced Protocols for Animal Transgenesis

Part of the book series: Springer Protocols Handbooks ((SPH))

  • 1638 Accesses

Abstract

Cryopreservation is the method of choice not only for archiving mouse mutant models at the end of an experiment, but also to create a security back up during their development. Three key factors need to be considered in cryopreservation: why, when, and how. In this chapter, we intend to discuss these factors and to provide an overview of current cryopreservation techniques. We will also refer to some experimental approaches that may become used more widely in the future. We will try to highlight advantages and disadvantages of each method with regard to investment in equipment, skills and technical limitations, and also review factors that compromise the efficiency in each approach. Finally, we will compile specific protocols for freezing mouse spermatozoa and embryos.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Knight J, Abbott A (2002) Full house. Nature 417:785–786

    Article  PubMed  CAS  Google Scholar 

  2. Ogonuki N, Mochida K, Miki H, Inoue K, Fray M, Iwaki T, Moriwaki K, Obata Y, Morozumi K, Yanagimachi R, Ogura A (2006) Spermatozoa and spermatids retrieved from frozen reproductive organs or frozen whole bodies of male mice can produce normal offspring. Proc Natl Acad Sci USA 103:13098–13103

    Article  PubMed  CAS  Google Scholar 

  3. Wakayama S, Ohta H, Hikichi T, Mizutani E, Iwaki T, Kanagawa O, Wakayama T (2008) Production of healthy cloned mice from bodies frozen at −20 degrees C for 16 years. Proc Natl Acad Sci USA 105:17318–17322

    Article  PubMed  CAS  Google Scholar 

  4. Garrick D, Fiering S, Martin DI, Whitelaw E (1998) Repeat-induced gene silencing in mammals. Nat Genet 18:56–59

    Article  PubMed  CAS  Google Scholar 

  5. Houdebine LM (2000) Transgenic animal bioreactors. Transgenic Res 9:305–320

    Article  PubMed  CAS  Google Scholar 

  6. Koetsier PA, Mangel L, Schmitz B, Doerfler W (1996) Stability of transgene methylation patterns in mice: position effects, strain specificity and cellular mosaicism. Transgenic Res 5:235–244

    Article  PubMed  CAS  Google Scholar 

  7. Whittingham DG, Leibo SP, Mazur P (1972) Survival of mouse embryos frozen to −196 degrees and −269 degrees C. Science 178:411–414

    Article  PubMed  CAS  Google Scholar 

  8. Critser JK, Mobraaten LE (2000) Cryopreservation of murine spermatozoa. ILAR J 41:197–206, National Research Council, Institute of Laboratory Animal Resources

    Article  PubMed  CAS  Google Scholar 

  9. Rall WF, Fahy GM (1985) Ice-free cryopreservation of mouse embryos at −196 degrees C by vitrification. Nature 313:573–575

    Article  PubMed  CAS  Google Scholar 

  10. Szell A, Shelton JN (1986) Sucrose dilution of glycerol from mouse embryos frozen rapidly in liquid nitrogen vapour. J Reprod Fertil 76:401–408

    Article  PubMed  CAS  Google Scholar 

  11. Leibo SP (1989) Equilibrium and nonequilibrium cryopreservation of embryos. Theriogenology 31:85–93

    Article  Google Scholar 

  12. Chupin D, De Reviers MM (1986) Quick freezing of rat embryos. Theriogenology 26:157–167

    Article  PubMed  CAS  Google Scholar 

  13. Abas Mazni O, Valdez CA, Takahashi Y, Hishinuma M, Kanagawa H (1990) Quick freezing of mouse embryos using ethylene glycol with lactose or sucrose. Anim Reprod Sci 22:161–169

    Article  Google Scholar 

  14. Kong IK, Lee SI, Cho SG, Cho SK, Park CS (2000) Comparison of open pulled straw (OPS) vs glass micropipette (GMP) vitrification in mouse blastocysts. Theriogenology 53:1817–1826

    Article  PubMed  CAS  Google Scholar 

  15. Tsang WH, Chow KL (2009) Mouse embryo cryopreservation utilizing a novel high-capacity vitrification spatula. Biotechniques 46:550–552

    Article  PubMed  CAS  Google Scholar 

  16. Peters DD, Marschall S, Mahabir E, Boersma A, Heinzmann U, Schmidt J, Hrabe de Angelis M (2006) Risk assessment of mouse hepatitis virus infection via in vitro fertilization and embryo transfer by the use of zona-intact and laser-microdissected oocytes. Biol Reprod 74:246–252

    Article  PubMed  CAS  Google Scholar 

  17. Schiewe MC, Hollifield VM, Kasbohm LA, Schmidt PM (1995) Embryo importation and cryobanking strategies for laboratory animals and wildlife species. Theriogenology 43:97–104

    Article  Google Scholar 

  18. Van Keuren ML, Saunders TL (2004) Rederivation of transgenic and gene-targeted mice by embryo transfer. Transgenic Res 13:363–371

    Article  PubMed  Google Scholar 

  19. Hogan B, Beddington R, Costantini F, Lacy E (1994) Manipulating the mouse embryo: a laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  20. Chatot CL, Ziomek CA, Bavister BD, Lewis JL, Torres I (1989) An improved culture medium supports development of random-bred 1-cell mouse embryos in vitro. J Reprod Fertil 86:679–688

    Article  PubMed  CAS  Google Scholar 

  21. Suzuki O, Asano T, Yamamoto Y, Takano K, Koura M (1996) Development in vitro of preimplantation embryos from 55 mouse strains. Reprod Fertil Dev 8:975–980

    Article  PubMed  CAS  Google Scholar 

  22. Byers SL, Payson SJ, Taft RA (2006) Performance of ten inbred mouse strains following assisted reproductive technologies (ARTs). Theriogenology 65:1716–1726

    Article  PubMed  Google Scholar 

  23. Munoz I, Rodriguez de Sadia C, Gutierrez A, Blanquez MJ, Pintado B (1994) Comparison of superovulatory response of mature outbred mice treated with FSH or PMSG and developmental potential of embryos produced. Theriogenology 41:907–914

    Article  PubMed  CAS  Google Scholar 

  24. Rall WF, Schmidt PM, Lin X, Brown SS, Ward AC, Hansen CT (2000) Factors affecting the efficiency of embryo cryopreservation and rederivatoin of rat and mouse models. ILAR J 41:221–227, National Research Council, Institute of Laboratory Animal Resources

    Article  PubMed  CAS  Google Scholar 

  25. Willoughby CE, Mazur P, Peter AT, Critser JK (1996) Osmotic tolerance limits and properties of murine spermatozoa. Biol Reprod 55:715–727

    Article  PubMed  CAS  Google Scholar 

  26. Tada N, Sato M, Yamanoi J, Mizorogi T, Kasai K, Ogawa S (1990) Cryopreservation of mouse spermatozoa in the presence of raffinose and glycerol. J Reprod Fertil 89:511–516

    Article  PubMed  CAS  Google Scholar 

  27. Sztein JM, Farley JS, Mobraaten LE (2000) In vitro fertilization with cryopreserved inbred mouse sperm. Biol Reprod 63:1774–1780

    Article  PubMed  CAS  Google Scholar 

  28. Nakagata N (2000) Cryopreservation of mouse spermatozoa. Mamm Genome 11:572–576

    Article  PubMed  CAS  Google Scholar 

  29. Ostermeier GC, Wiles MV, Farley JS, Taft RA (2008) Conserving, distributing and managing genetically modified mouse lines by sperm cryopreservation. PLoS ONE 3:e2792

    Article  PubMed  Google Scholar 

  30. Takeo T, Hoshii T, Kondo Y, Toyodome H, Arima H, Yamamura K, Irie T, Nakagata N (2008) Methyl-beta-cyclodextrin improves fertilizing ability of C57BL/6 mouse sperm after freezing and thawing by facilitating cholesterol efflux from the cells. Biol Reprod 78:546–551

    Article  PubMed  CAS  Google Scholar 

  31. Mahabir E, Bulian D, Needham J, Schmidt J (2009) Lack of transmission of mouse minute virus (MMV) from in vitro-produced embryos to recipients and pups due to the presence of cumulus cells during the in vitro fertilization process. Biol Reprod 81(3):531–538

    Article  PubMed  CAS  Google Scholar 

  32. Janus LM, Smoczek A, Hedrich HJ, Bleich A (2009) Risk assessment of minute virus of mice transmission during rederivation: detection in reproductive organs, gametes, and embryos of mice after in vivo infection. Biol Reprod 81(5):1010–1015

    Article  PubMed  CAS  Google Scholar 

  33. Bielanski A, Bergeron H, Lau PC, Devenish J (2003) Microbial contamination of embryos and semen during long term banking in liquid nitrogen. Cryobiology 46:146–152

    Article  PubMed  CAS  Google Scholar 

  34. Bath ML (2003) Simple and efficient in vitro fertilization with cryopreserved C57BL/6J mouse sperm. Biol Reprod 68:19–23

    Article  PubMed  CAS  Google Scholar 

  35. Pintado B, Hourcade JD, Pérez-Crespo M, Gutiérrez-Adán A (2008) Intraoviductal insemination with frozen C57Bl/6J sperm increases fertility rate compared to standard IVF. Transgenic Res 17:1017

    Google Scholar 

  36. Wakayama T, Whittingham DG, Yanagimachi R (1998) Production of normal offspring from mouse oocytes injected with spermatozoa cryopreserved with or without cryoprotection. J Reprod Fertil 112:11–17

    Article  PubMed  CAS  Google Scholar 

  37. Snow M, Cox SL, Jenkin G, Trounson A, Shaw J (2002) Generation of live young from xenografted mouse ovaries. Science 297:2227

    Article  PubMed  CAS  Google Scholar 

  38. Wakayama S, Kishigami S, Wakayama T (2009) Cloning of ES cells and mice by nuclear transfer. Meth Mol Biol 530:251–265, Clifton, NJ

    Article  CAS  Google Scholar 

  39. IETS (1998) Manual of the International Embryo Transfer Society: a procedural guide and general information for the use of embryo transfer technology, emphasizing sanitary precautions, 3rd edn. IETS, Champaign, IL

    Google Scholar 

  40. Gutierrez A, Garde J, Artiga CG, Munoz I, Pintado B (1993) In vitro survival of murine morulae after quick freezing in the presence of chemically defined macromolecules and different cryoprotectants. Theriogenology 39:1111–1120

    Article  PubMed  CAS  Google Scholar 

  41. Sztein JM, Farley JS, Young AF, Mobraaten LE (1997) Motility of cryopreserved mouse spermatozoa affected by temperature of collection and rate of thawing. Cryobiology 35:46–52

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Transgenic Service. Centro Nacional de Biotecnología, CSIC, C/Darwin 3, 28040, Madrid, Spain

    B. Pintado

  2. Dpto Reproducción Animal, INIA, Ctra de La Coruña Km- 5,9, 28040, Madrid, Spain

    J. Hourcade

Authors
  1. B. Pintado
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Hourcade
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to B. Pintado .

Editor information

Editors and Affiliations

  1. Div. Biology, California Institute of Technology, Pasadena, 91125, California, USA

    Shirley Pease

  2. Transgenic Animal Model Core, University of Michigan, Ann Arbor, 48109-5674, Michigan, USA

    Thomas L. Saunders

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer-Verlag Berlin Heidelberg

About this protocol

Cite this protocol

Pintado, B., Hourcade, J. (2011). Cryopreservation. In: Pease, S., Saunders, T. (eds) Advanced Protocols for Animal Transgenesis. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-20792-1_23

Download citation

  • DOI: https://doi.org/10.1007/978-3-642-20792-1_23

  • Published:

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-20791-4

  • Online ISBN: 978-3-642-20792-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation