Skip to main content
SpringerLink
Log in

Evaluation of recombinant human transferrin (DeltaFerrinTM) as an iron chelator in serum-free media for mammalian cell culture

  • Original Paper
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

DeltaFerrinTM, a yeast-derived recombinant human transferrin produced by Delta Biotechnology Ltd. (Nottingham UK), was found to be a suitable replacement for holo human transferrin in serum-free culture media of the MDCK cell line (chosen because of its transferrin dependence) in short-term screening assays. Long-term subculture was achieved with DeltaFerrinTM supporting growth equivalent to that of holo human transferrin. DeltaFerrinTM and a selection of chemical iron chelators were found in short-term assays to be equivalent to holo human transferrin in supporting growth of MDCK, BHK-21-PPI-C16 and Vero-PPI. In long-term subcultures, however, only DeltaFerrinTM was found to support cell growth in a manner essentially equivalent to holo human transferrin in all three cell lines. For both BHK and Vero variants tested, recombinant preproinsulin production was unaltered by replacing holo human transferrin with DeltaFerrinTM. As such, this is the first report of a recombinant human transferrin produced under animal-free conditions that can act as a universal iron chelator for cells grown in serum-free media (SFM).

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Barnes D (1987) Serum-free animal cell culture. BioTechniques 5:534–542

    Article  CAS  Google Scholar 

  • Bradshaw GL, Sato GH, McClure DB, Dubes GR (1994) The growth requirements of BHK-21 in serum-free culture. J Cell Physiol 114:215–221

    Article  Google Scholar 

  • Castle P, Robertson JS (1999) Animal sera, animal sera derivatives and substitutes used in the manufacture of pharmaceuticals: viral safety and regulatory aspects. Dev Biol Standard 99:191–196

    CAS  Google Scholar 

  • Chu L, Robinson DK (2001) Industrial choices for protein production by large-scale cell culture. Curr Opin Biotechnol 12:180–187

    Article  CAS  Google Scholar 

  • Conrad ME, Umbreit JN, Morre EG (1994) Iron absorption and transport. Am J Med Sci 318:213–229

    Article  Google Scholar 

  • Cruz HJ, Moreira JL, Stacey G, Dias EM, Hayes K, Looby D, Griffiths B, Carrondo MJT(1998) Adaptation of BHK cells producing a recombinant protein to serum-free media and protein-free medium. Cytotechnology 26:59–64

    Article  CAS  Google Scholar 

  • Darfler FJ (1990) A protein-free medium for the growth of hybridomas and other cells of the immune system. In Vitro Cell Develop Biol 26:769–778

    Article  CAS  Google Scholar 

  • Fitzsimmons JJ, Sanjal A, Gonzalez C, Fukumoto T, Clemens VR, O’Driscoll SW, Reinholz GG (2004) Serum-free media for periosteal chondrogenesis in vitro. J Orthop Res 22:716–725

    Article  CAS  Google Scholar 

  • Gammell P, O’Driscoll L, Clynes M (2003). Characterisation of BHK-21-PPI-C16-21 cells engineered to secrete human insulin. Cytotechnology 41:11–21

    Article  CAS  Google Scholar 

  • Jayme DW (1999) An animal origin perspective of common constituents of serum-free medium formulations. Dev Biol Standard 99:181–187

    CAS  Google Scholar 

  • Kallel H, Jouini A, Majoul S, Rourou S (2002) Evaluation of various serum and animal protein free media for the production of a veterinary rabies vaccine in BHK-21 cells. J Biotechnol 95:195–204

    Article  CAS  Google Scholar 

  • Keenan J, Clynes M (1996) Replacement of transferrin by simple iron compounds for MDCK cells grown and subcultured in serum-free medium. In Vitro Cell Develop Biol 32:451–453

    Article  CAS  Google Scholar 

  • Laskey J, Webb I, Schulman H, Ponka P (1988) Evidence that transferrin supports cell proliferation by supplying iron for DNA synthesis. Exp Cell Res 176:87–95

    Article  CAS  Google Scholar 

  • Litwin J (1992) The growth of Vero cells in suspension as cell aggregates in serum-free medium. Cytotechnology 10:169–174

    Article  CAS  Google Scholar 

  • Martin A, Clynes M (1993) Comparison of 5 microplate colorimetric assays for in vitro cytotoxicity testing and cell proliferation assays. Cytotechnology 11:49–58

    Article  CAS  Google Scholar 

  • Metcalfe H, Field RP, Froud SJ (1994) The use of 2-hydroxy-2,4,6-cycloheptarin-1-one (Tropolone) as a replacement for transferrin. In: Spier RE, Griffiths JB, Berthold W (eds) Animal cell technololgy: products of today, prospects for tomorrow. Butterwirth-Heinemann, Oxford/U.K., pp 88–90

    Google Scholar 

  • Merten O-W (2002) Development of serum-free media for cell growth and production of viruses/viral vectors – safety issues of animal products used in serum-free media. Dev Biol 111:233–257

    CAS  Google Scholar 

  • Merten O-W, Hannoun C, Manuguerra JC, Ventre F, Petres S (1996) Production of influenza virus in cell␣cultures for vaccine preparation. In: Cohen S, Shafferman A (eds) Novel strategies in design and production of vaccines. Plenum Press, New York, USA, pp 141–151

    Google Scholar 

  • Merten O-W, Kallel H, Manuguerra JC, Tardy-Panit M, Crainic R, Delpeyroux F, Van der Werf S, Perrin P (1999) The new medium MDSS2N, free of any animal protein supports cell growth and production of various viruses. Cytotechnology 30:191–201

    Article  CAS  Google Scholar 

  • Neumannova V, Richardson DR, Kriegerbeckova V, Kovar J (1995) Growth of human tumor cell lines in transferrin-free low iron medium. In Vitro Cell Develop Biol 31:625–632

    Article  CAS  Google Scholar 

  • O’Driscoll L, Gammell P, Clynes M (2002) Engineering Vero-PPI cells to secrete human insulin. In Vitro Cell Develop Biol 38:146–153

    Article  CAS  Google Scholar 

  • Okamoto T, Tani R, Yabamoto M, Sakamoto A, Takada K, Sato GH, Sato JG (1996) Effects of insulin and transferrin on the generation of lymphokine-activated killer cells in serum-free medium. J Immunol Method 195:7–14

    Article  CAS  Google Scholar 

  • Perrin P, Malhusudana S, Gontier-Jallet C, Petres S, Tordo N, Merten O-W (1995) An experimental rabies vaccine produced with a new BHK-21 suspension culture process: use of serum-free medium and perfusion-reactor system. Vaccine 13:1244–1250

    Article  CAS  Google Scholar 

  • Richardson DR, Ponka P (1997) The molecular mechanism of the metabolism and transport of iron in normal and neoplastic cells. Biochim Biophys Acta 1331:1–40

    CAS  Google Scholar 

  • Salis C, Goedelmann CJ, Pasquini JM, Soto EF, Setton-Avruj CP (2002) Holo transferrin but not apo transferrin prevents Schwann cell de-differentiation in culture. Dev Neurosci 24:214–221

    Article  CAS  Google Scholar 

  • Sanders EJ, Cheung E (1988) Transferrin and iron requirements of embryonic mesoderm cells cultured in hydrated collagen matrices. In Vitro Cell Develop Biol 24:581–587

    Article  CAS  Google Scholar 

  • Shintani N, Kohgo Y, Kato J, Kondo H, Fujikawa K, Miyazaki E, Niitsu Y (1994) Expression and extracellular release of transferrin receptors during peripheral erythroid progenitor cell differentiation in liquid culture. Blood 83:1209–1215

    CAS  Google Scholar 

  • Taub M, Chuman L, Saier MH, Sato G (1979) Growth of MDCK in hormonally defined serum-free medium. Proc Natl Acad Sci USA 76:3338–3342

    Article  CAS  Google Scholar 

  • Vyhlidal C, Li X, Safe S (2002) Estrogen regulation of transferrin gene expression in MCF-7 human breast cancer cells. J Mol Enodcrinol 29:305–317

    Article  CAS  Google Scholar 

  • Wong VVT, Ho KW, Yap MGS (2004) Evaluation of insulin-mimetic trace metals as insulin replacements in mammalian cell cultures. Cytotechnology 45:107–115

    Article  CAS  Google Scholar 

  • Yabe N, Kato M, Matsuya Y, Yamane I, Iizuka M, Takayoshi H, Suzuki K (1987) Role of iron chelators in growth-promoting effect on mouse hybridoma cells in a chemically defined medium. In Vitro Cell Develop Biol 23:815–820

    Article  CAS  Google Scholar 

  • Yamada K, Ikeda I, Sughara T, Hashizume S, Shirahata S, Murakami H (1990) Stimulation of proliferation and immunoglobulin M production by lactoferrin in human–human and mouse–mouse hybridoma cultures in serum-free conditions. Cytotechnology 3:123–131

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. National Institute for Cellular Biotechnology, Dublin City University, Dublin, Ireland

    Joanne Keenan, Lorraine O’Driscoll, Patrick Gammell & Martin Clynes

  2. Delta Biotechnology Ltd., Nottingham, UK

    Dermot Pearson

Authors
  1. Joanne Keenan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dermot Pearson
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lorraine O’Driscoll
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Patrick Gammell
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Martin Clynes
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Joanne Keenan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Keenan, J., Pearson, D., O’Driscoll, L. et al. Evaluation of recombinant human transferrin (DeltaFerrinTM) as an iron chelator in serum-free media for mammalian cell culture. Cytotechnology 51, 29–37 (2006). https://doi.org/10.1007/s10616-006-9011-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-006-9011-x

Keywords

Search

Navigation