- Published:
Serum-free cryopreservation of five mammalian cell lines in either a pelleted or suspended state
Biological Procedures Online volume 6, pages 61–66 (2004)
Abstract
Herein we have explored two practical aspects of cryopreserving cultured mammalian cells during routine laboratory maintenance. First, we have examined the possibility of using a serum-free, hence more affordable, cryopreservative. Using five mammalian lines (Crandell Feline Kidney, MCF7, A72, WI 38 and NB324K), we found that the serum-free alternative preserves nearly as efficiently as the serum-containing preservatives. Second, we compared cryostorage of those cells in suspended versus a pellet form using both aforementioned cryopreservatives. Under our conditions, cells were in general recovered equally well in a suspended versus a pellet form.
References
Polge AS, Smith AU, Parks AS. Survival of spermatozoa after dehydration and vitrification at low temperature. Nature 1949; 164:666.
Mazur P. Cryobiology: The freezing of biological systems. Science 1970; 168:939–949.
Mazur P, Leibo SP, Chu EHY. A two-factor hypothesis of freezing injury. Exp Cell Res 1971; 72:345–355.
Lovelock JE, Bishop MH. Prevention of freezing damage to living cells by dimethyl-sulphoxide. Nature 1959; 183:1394–1395.
Beckmann J, Korber CH, Rau G, Hubel A, Cravalho EG. Redefining cooling rates in terms of ice front velocity and thermal gradient: first evidence of relevance to freezing injury of lymphocytes. Cryobiology 1990; 27:279–287.
Kelbe RJ, Mancuso MG. Identification of new cryoprotective agents for cultured mammalian cells. In Vitro 1983; 19:167–170.
Lim JM, Ko JJ, Hwang WS, Chung HM, Niwa K. Development of in vitro matured oocytes after cryopreservation with different cryoprotectants. Theriogenology 1999; 51:1303–1310.
Hubel A, Cravalho EG, Nunner B, Korber C. Survival of directionaly solidified B-lymphoblasts under various crystal growth conditions. Cryobiology 1992; 29:183–198.
Acker JP, McGann LE. Cell-cell contact affects membrane integrity after intracellular freezing. Cryobiology 2000; 40:54–63.
Acker JP, Larese A, Yang H, Petrenko A, McGrann LE. Intracellular ice formation is affected by cell interactions. Cryobiology 1999; 38:363–371.
Armitage WJ, Juss BK. Assembly of intercellular junctions in epithelial cell monolayers following exposure to cryoprotectants. Cryobiology 2000; 41:58–65.
Crandell RA, Fabricant CG, Nelson-Rees WA. Development, characterization, and viral susceptibility of a feline (Felis catus) renal cell line (CRFK). In Vitro 1973; 9:176–185.
Spitzer AL, Maxwell F, Corsini J, Maxwell IH. Species specificity for transduction of cultured cells by a recombinant LuIII rodent parvovirus genome encapsidated by canine parvovirus or feline panleukopenia virus. J Gen Virol 1996; 77:1787–1792.
Spitzer AL, Parrish CR, Maxwell IH. Tropic determinant for canine parvovirus and feline panleukopenia virus functions through the capsid protein VP2. J Gen Virol 1997; 78:925–928.
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.
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.
Parrish CR, Aquadro CF, Carmichael LE. Canine host range and a specific epitope map along with variant sequences in the capsid protein gene of canine parvovirus and related feline, mink, and raccoon parvoviruses. Virology 1988; 166:293–307.
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.
Corsini J, Maxwell F, Maxwell IH. Storage of various cell lines at −70C or −80C in multi-well plates while attached to the substratum. Biotechniques 2002; 33:1–3.
Corsini J, Maxwell IH, Maxwell F, Carlson JO. Expression of parvovirus LuIII NS1 from a Sindbis replicon for production of LuIII-luciferase transducing virus. Virus Res 1996; 46:95–104.
Hayflick L, Moorhead PS. The serial cultivation of human diploid cell strains. Exp Cell Res 1961; 25: 585–621.
Hayflick L, Plotkin SA, Norton TW, Koprowski IH. Preparation of poliovirus vaccines in a human fetal diploid cell strain. Am J Hyg 1962; 75:240–258.
Hayflick L. The limited in vitro lifetime of human diploid cell strains. Exp Cell Res 1965; 37: 614–636.
Author information
Authors and Affiliations
Corresponding author
Additional information
Published: April 7, 2004.
Rights and permissions
About this article
Cite this article
Corsini, J., Hacker, C. & Bare, C. Serum-free cryopreservation of five mammalian cell lines in either a pelleted or suspended state. Biol. Proced. Online 6, 61–66 (2004). https://doi.org/10.1251/bpo73
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1251/bpo73