Skip to main content

Rapid cryopreservation of five mammalian and one mosquito cell line at −80°C while attached to flasks in a serum free cryopreservative


Cell culturing, and the requisite storage of cell lines at ultra-low temperatures, is used in most laboratories studying or using eukaryotic proteomics, genomics, microarray, and RNA technologies. In this study we have observed that A72(dog), CRFK(cat), NB324K(human), MCF7(human), WI38(human), and C636(mosquito) cells were effectively cryopreserved at −80°C while attached to the substratum of 25cm2 tissue culture flasks. This was accomplished using a serum free crypreservative recently developed by Corsini and co-workers. The technique allows for significant savings of time and money in laboratories that rapidly process numerous cell lines.


  1. Corsini J, Maxwell F, Maxwell IH. Storage of various cell lines at −70°C or −80°C in multi-well plates while attached to the substratum. BioTechniques 2002; 33:1–3.

    Google Scholar 

  2. Wells DE, Price PJ. Simple rapid methods for freezing hybridomas in 96-well microculture plates. J Immun Methods 1983; 59:49–52.

    Article  CAS  Google Scholar 

  3. De Leij L, Poppema S, The TH. Cryopreservation of newly formed hybridomas. J Immun Methods 1983;62:69–72.

    Article  Google Scholar 

  4. Corsini J, Hacker C, Bare C. Serum-free cryopreservation of five mammalian cell lines in either a pelleted or a suspended state. Biol Proc Online 2004; 6:61–66.

    Article  CAS  Google Scholar 

  5. Igarashi A. Isolation of a Singh’s Aedes albopictus cell clone sensitive to Dengue and Chikungunya viruses. J Gen Virol 1978; 40: 531–544.

    Article  PubMed  CAS  Google Scholar 

  6. Shein HM, Enders JF. Multiplication and cytopathogenicity of simian vacuolating virus in cultures of human tissues. Proc Soc Exp Biol Med 1962; 109:495–500.

    PubMed  CAS  Google Scholar 

  7. Crandell RA, Fabricant CG, Nelson-Rees WA. Development, characterization, and viral susceptibility of a feline (Felis catis) renal cell line (CRFK). In Vitro 1973; 9:176–185.

    Article  PubMed  CAS  Google Scholar 

  8. Binn LN, Marchwicki RH, Stephenson EH. Establishment of a canine cell line: derivation, characterization, and viral spectrum. Am J Vet Res 1980; 41:855–860.

    PubMed  CAS  Google Scholar 

  9. Sugarman BJ, Aggarwal BB, Hass PE, Figari IS, Palladino IA, Jr. Shepard HM. Recombinant human tumor necrosis factor-alpha: effects on proliferation of normal and transformed cells in vitro. Science 1985; 230: 943–945.

    Article  PubMed  CAS  Google Scholar 

  10. Hayflick L, Moorhead PS. The serial cultivation of human diploid strains. Exp Cell Res 1961; 25:585–621.

    Article  Google Scholar 

  11. Hayflick L, Plotkin SA, Norton TW, Koprowski IH. Preparation of poliovirus vaccines in a human fetal diploid strain. Am J Hyg 1962; 75:240–258.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations


Corresponding author

Correspondence to Joe Corsini.

Rights and permissions

Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution License ( ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Reprints and Permissions

About this article

Cite this article

Corsini, J., Mann, E. Rapid cryopreservation of five mammalian and one mosquito cell line at −80°C while attached to flasks in a serum free cryopreservative. Biol. Proced. Online 7, 26–30 (2005).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI:

Indexing terms

  • Cryopreservation
  • Culture Media, Serum-Free
  • Cells, Cultured