• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of cytotechspringer.comThis journalToc AlertsSubmit OnlineOpen Choice
Cytotechnology. May 2006; 51(1): 29–37.
Published online Aug 3, 2006. doi:  10.1007/s10616-006-9011-x
PMCID: PMC3449479

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


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).

Keywords: Cell culture, Iron chelator, DeltaFerrinTM, Recombinant human transferrin, Serum-free media

Full Text

The Full Text of this article is available as a PDF (323K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Barnes D. Serum-free animal cell culture. BioTechniques. 1987;5:534–542. doi: 10.1038/nbt0687-534. [Cross Ref]
  • Bradshaw GL, Sato GH, McClure DB, Dubes GR. The growth requirements of BHK-21 in serum-free culture. J Cell Physiol. 1994;114:215–221. doi: 10.1002/jcp.1041140211. [PubMed] [Cross Ref]
  • Castle P, Robertson JS. Animal sera, animal sera derivatives and substitutes used in the manufacture of pharmaceuticals: viral safety and regulatory aspects. Dev Biol Standard. 1999;99:191–196. [PubMed]
  • Chu L, Robinson DK. Industrial choices for protein production by large-scale cell culture. Curr Opin Biotechnol. 2001;12:180–187. doi: 10.1016/S0958-1669(00)00197-X. [PubMed] [Cross Ref]
  • Conrad ME, Umbreit JN, Morre EG. Iron absorption and transport. Am J Med Sci. 1994;318:213–229. doi: 10.1097/00000441-199910000-00002. [PubMed] [Cross Ref]
  • Cruz HJ, Moreira JL, Stacey G, Dias EM, Hayes K, Looby D, Griffiths B, Carrondo MJT. Adaptation of BHK cells producing a recombinant protein to serum-free media and protein-free medium. Cytotechnology. 1998;26:59–64. doi: 10.1023/A:1007951813755. [PMC free article] [PubMed] [Cross Ref]
  • Darfler FJ. A protein-free medium for the growth of hybridomas and other cells of the immune system. In Vitro Cell Develop Biol. 1990;26:769–778. doi: 10.1007/BF02623618. [PubMed] [Cross Ref]
  • Fitzsimmons JJ, Sanjal A, Gonzalez C, Fukumoto T, Clemens VR, O’Driscoll SW, Reinholz GG. Serum-free media for periosteal chondrogenesis in vitro. J Orthop Res. 2004;22:716–725. doi: 10.1016/j.orthres.2003.10.020. [PubMed] [Cross Ref]
  • Gammell P, O’Driscoll L, Clynes M. Characterisation of BHK-21-PPI-C16–21 cells engineered to secrete human insulin. Cytotechnology. 2003;41:11–21. doi: 10.1023/A:1024296220592. [PMC free article] [PubMed] [Cross Ref]
  • Jayme DW. An animal origin perspective of common constituents of serum-free medium formulations. Dev Biol Standard. 1999;99:181–187. [PubMed]
  • Kallel H, Jouini A, Majoul S, Rourou S. Evaluation of various serum and animal protein free media for the production of a veterinary rabies vaccine in BHK-21 cells. J Biotechnol. 2002;95:195–204. doi: 10.1016/S0168-1656(02)00009-3. [PubMed] [Cross Ref]
  • Keenan J, Clynes M. Replacement of transferring by simple iron compounds for MDCK cells grown and subcultured in serum-free medium. In Vitro Cell Develop Biol. 1996;32:451–453. doi: 10.1007/BF02723044. [PubMed] [Cross Ref]
  • Laskey J, Webb I, Schulman H, Ponka P. Evidence that transferrin supports cell proliferation by supplying iron for DNA synthesis. Exp Cell Res. 1988;176:87–95. doi: 10.1016/0014-4827(88)90123-1. [PubMed] [Cross Ref]
  • Litwin J. The growth of Vero cells in suspension as cell aggregates in serum-free medium. Cytotechnology. 1992;10:169–174. doi: 10.1007/BF00570893. [PubMed] [Cross Ref]
  • Martin A, Clynes M. Comparison of 5 microplate colorimetric assays for in vitro cytotoxicity testing and cell proliferation assays. Cytotechnology. 1993;11:49–58. doi: 10.1007/BF00749057. [PubMed] [Cross Ref]
  • Metcalfe H, Field RP, Froud SJ. The use of 2-hydroxy-2,4,6-cycloheptarin-1-one (Tropolone) as a replacement for transferring. In: Spier RE, Griffiths JB, Berthold W, editors. Animal cell technololgy: products of today, prospects for tomorrow. Oxford/U.K.: Butterwirth-Heinemann; 1994. pp. 88–90.
  • Merten O-W. 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. 2002;111:233–257. [PubMed]
  • Merten OW, Hannoun C, Manuguerra JC, Ventre F, Petres S. Production of influenza virus in cell cultures for vaccine preparation. In: Cohen S, Shafferman A, editors. Novel strategies in design and production of vaccines. New York, USA: Plenum Press; 1996. pp. 141–151.
  • Merten O-W, Kallel H, Manuguerra JC, Tardy-Panit M, Crainic R, Delpeyroux F, Werf S, Perrin P. The new medium MDSS2N, free of any animal protein supports cell growth and production of various viruses. Cytotechnology. 1999;30:191–201. doi: 10.1023/A:1008021317639. [PMC free article] [PubMed] [Cross Ref]
  • Neumannova V, Richardson DR, Kriegerbeckova V, Kovar J. Growth of human tumor cell lines in transferrin-free low iron medium. In Vitro Cell Develop Biol. 1995;31:625–632. doi: 10.1007/BF02634316. [PubMed] [Cross Ref]
  • O’Driscoll L, Gammell P, Clynes M. Engineering Vero-PPI cells to secrete human insulin. In Vitro Cell Develop Biol. 2002;38:146–153. doi: 10.1290/1071-2690(2002)038<0146:EVCTSH>2.0.CO;2. [PubMed] [Cross Ref]
  • Okamoto T, Tani R, Yabamoto M, Sakamoto A, Takada K, Sato GH, Sato JG. Effects of insulin and transferrin on the generation of lymphokine-activated killer cells in serum-free medium. J Immunol Method. 1996;195:7–14. doi: 10.1016/0022-1759(96)00081-6. [PubMed] [Cross Ref]
  • Perrin P, Malhusudana S, Gontier-Jallet C, Petres S, Tordo N, Merten O-W. An experimental rabies vaccine produced with a new BHK-21 suspension culture process: use of serum-free medium and perfusion-reactor system. Vaccine. 1995;13:1244–1250. doi: 10.1016/0264-410X(94)00022-F. [PubMed] [Cross Ref]
  • Richardson DR, Ponka P. The molecular mechanism of the metabolism and transport of iron in normal and neoplastic cells. Biochim Biophys Acta. 1997;1331:1–40. [PubMed]
  • Salis C, Goedelmann CJ, Pasquini JM, Soto EF, Setton-Avruj CP. Holo transferrin but not apo transferrin prevents Schwann cell de-differentiation in culture. Dev Neurosci. 2002;24:214–221. doi: 10.1159/000065695. [PubMed] [Cross Ref]
  • Sanders EJ, Cheung E. Transferrin and iron requirements of embryonic mesoderm cells cultured in hydrated collagen matrices. In Vitro Cell Develop Biol. 1988;24:581–587. doi: 10.1007/BF02629094. [PubMed] [Cross Ref]
  • Shintani N, Kohgo Y, Kato J, Kondo H, Fujikawa K, Miyazaki E, Niitsu Y. Expression and extracellular release of transferrin receptors during peripheral erythroid progenitor cell differentiation in liquid culture. Blood. 1994;83:1209–1215. [PubMed]
  • Taub M, Chuman L, Saier MH, Sato G. Growth of MDCK in hormonally defined serum-free medium. Proc Natl Acad Sci USA. 1979;76:3338–3342. doi: 10.1073/pnas.76.7.3338. [PMC free article] [PubMed] [Cross Ref]
  • Vyhlidal C, Li X, Safe S. Estrogen regulation of transferrin gene expression in MCF-7 human breast cancer cells. J Mol Enodcrinol. 2002;29:305–317. doi: 10.1677/jme.0.0290305. [PubMed] [Cross Ref]
  • Wong VVT, Ho KW, Yap MGS. Evaluation of insulin-mimetic trace metals as insulin replacements in mammalian cell cultures. Cytotechnology. 2004;45:107–115. doi: 10.1007/s10616-004-6173-2. [PMC free article] [PubMed] [Cross Ref]
  • Yabe N, Kato M, Matsuya Y, Yamane I, Iizuka M, Takayoshi H, Suzuki K. Role of iron chelators in growth-promoting effect on mouse hybridoma cells in a chemically defined medium. In Vitro Cell Develop Biol. 1987;23:815–820. doi: 10.1007/BF02620959. [PubMed] [Cross Ref]
  • Yamada K, Ikeda I, Sughara T, Hashizume S, Shirahata S, Murakami H. Stimulation of proliferation and immunoglobulin M production by lactoferrin in human–human and mouse–mouse hybridoma cultures in serum-free conditions. Cytotechnology. 1990;3:123–131. doi: 10.1007/BF00143674. [PubMed] [Cross Ref]

Articles from Cytotechnology are provided here courtesy of Springer Science+Business Media B.V.


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...