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Cryopreservation of Primary Animal Cell Cultures

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Cryopreservation and Freeze-Drying Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 368))

Abstract

Cells isolated directly from tissues (primary cultures) have many applications in research and applications in the fields of toxicology, pharmacology, and virology. Where such preparations remain the only scientific option to achieve necessary results it is still possible to refine and reduce the use of animal and human sources of the tissue by cryopreservation of the primary cells for later use, this reduces the need for fresh tissues and enables improvements in standardization through the ability to provide the same cell preparation at different times and to different laboratories. The methods described provide options for adherent cultures as monolayers, harvested cell suspensions, and also lymphocytes isolated from peripheral blood.

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References

  1. Russell, W. M. S. and Burch, R. L. (1959) The Principles of Humane Experimental Techniques. Methuen, London, UK.

    Google Scholar 

  2. Milstein, J., Grachev, V., Padilla, A., and Griffiths, E. (1996) WHO activities towards the three Rs in the development and control of biological medicines. Dev. Biol. Stand. 86, 31–39.

    Google Scholar 

  3. Stacey, G. N., Doyle, A., and Tyrrel, D. (1998) Source materials. In: Safety in Cell and Tissue Culture, (Stacey, G. N., Doyle, A., and Hambleton, P. H., eds.), Kluwer Academic Publishers, Dordrecht, Germany, pp. 1–25.

    Google Scholar 

  4. Coecke, S., Balls, M., Bowe, G., et al. (2005) Guidance on good cell culture practice: A report of the second ECVAM task force on good cell culture practice. ATLA 33, 1–27.

    Google Scholar 

  5. Thomson, J., Itskovitz-Eldor, J., Shapiro, S., et al. (1998) Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147.

    Article  CAS  Google Scholar 

  6. Terasima, T. and Yasukawa, M. (1977) Dependence of freeze-thaw damage on growth phase and cell cycle. Cryobiology 14, 379–381.

    Article  CAS  Google Scholar 

  7. Boyum, A. (1968) Separation of leukocytes from blood and bone marrow. Scand. J. Clin. Lab. Invest. 21, 77.

    Article  CAS  Google Scholar 

  8. Stacey, G. N. (1994) Chick embryo fibroblast preparation. In: Cell and Tissue Culture: Laboratory Procedures, (Doyle, A. and Griffiths, J.B., eds.), John Wiley and Sons, Chichester, UK, pp. 1.1.

    Google Scholar 

  9. Dowall, S. (1997) Cryopreservation of primary monkey kidney cells. Report submitted as part of the BTEC HNC in Applied Biology, Health Protection Agency, Salisbury, UK.

    Google Scholar 

  10. Freshney, I. R. (1994) Measurement of viability and cytotoxicity. In: Culture of Animal Cells: a Manual of Basic Techniques, (Freshney, I. R., ed.), Wiley Liss, New York, pp. 288–289.

    Google Scholar 

  11. Patterson, M. K. (1979) Measurement of growth and viability of cells in culture. Meth. Enzymol. 58, 141–152.

    Article  Google Scholar 

  12. Stacey, G. N. (2002) Standardisation of cell lines. Dev. Biologicals 111, 259–272.

    CAS  Google Scholar 

  13. Stacey, G. N. and Doyle, A. (2000) Cell banking. In: Encyclopedia of Cell Technology, (Spier, R., editor in chief), Wiley Interscience, New York, pp. 293–293.

    Google Scholar 

  14. Stacey, G. N. and Stacey, A. R. (1999) Routine quality control testing of cell cultures. In: Antiviral Methods and Protocols, (Kinchington, D. and Schinazi, R. F., eds.), Humana Press, Totowa, NJ, pp. 27–40.

    Chapter  Google Scholar 

  15. European Pharmacopoeia. (2004) European Pharmacopoeia section 2.6.1 (Sterility) and 2.6.7 (Mycoplasma), Maisonneuve SA, Sainte Ruffin, France.

    Google Scholar 

  16. Doblhoff-dier, O. and Stacey, G. N. (2000) Cell lines: applications and biosafety. In: Biological Safety Principles and Practices, 3rd ed., (Fleming, D. O. and Hunt, D. L., eds.), ASM Press, Washington, DC, pp. 221–240.

    Google Scholar 

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© 2007 Humana Press Inc., Totowa, NJ

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Stacey, G.N., Dowall, S. (2007). Cryopreservation of Primary Animal Cell Cultures. In: Day, J.G., Stacey, G.N. (eds) Cryopreservation and Freeze-Drying Protocols. Methods in Molecular Biology™, vol 368. Humana Press. https://doi.org/10.1007/978-1-59745-362-2_19

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  • DOI: https://doi.org/10.1007/978-1-59745-362-2_19

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-377-0

  • Online ISBN: 978-1-59745-362-2

  • eBook Packages: Springer Protocols

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