Skip to main content
Cytotechnology logoLink to Cytotechnology
. 2002 Nov;40(1-3):3–12. doi: 10.1023/A:1023993400608

Sericin, a protein derived from silkworms, accelerates the proliferation of several mammalian cell lines including a hybridoma

Satoshi Terada 1,, Taeko Nishimura 1, Masahiro Sasaki 2, Hideyuki Yamada 2, Masao Miki 1
PMCID: PMC3449532  PMID: 19003099

Abstract

Sericin, a constituent of the silkworm cocoon, was added to the culture of four mammalian cell lines: murine hybridoma 2E3-O,human hepatoblastoma HepG2, human epithelial HeLa and human embryonal kidney 293 cells. The proliferation of all cell lineswas accelerated in the presence of sericin. The hybridoma cellline was further studied. The 2E3-O cell line was so well adapted to serum-free medium that both the proliferation rate and maximum cell density in serum-free ASF103 medium were higher than in RPMI medium supplemented with all lots of FBS tested, and this proliferation was stimulated by the addition of sericin in a dose-dependent manner. Stimulation was observed at sericin concentrations from 0.01 to 0.1 %, although 1% sericin was severely harmful to the culture. In comparison with bovine serum albumin (BSA), a widely used supplement in serum-free medium, sericin had an equivalent effect on the proliferation of the hybridomas and sericin additively stimulated the proliferation with BSA. Although heat easily denatures and inactivates most proteins, the activity of sericin was not affected by autoclaving. In a similar manner to the silkworm-derived sericin, recombinant sericin synthesized in E. coli also stimulated the hybridoma proliferation, irrespective of whether it was autoclaved or filtered. Since BSA is obtained from bovine serum and the risk of infections such as bovine spongiform encephalopathy cannot be eradicated, sericin derived from insects could be a preferable culture medium supplement for stimulating the proliferation of mammalian cells.

Keywords: HeLa, HepG2, hybridoma, proliferation, sericin, serum-free medium

Full Text

The Full Text of this article is available as a PDF (146.8 KB).

References

  1. Chang TH, Steplewski Z, Koprowski H. Production of monoclonal antibodies in serum free medium. J Immunol Meth. 1980;39:369–375. doi: 10.1016/0022-1759(80)90237-9. [DOI] [PubMed] [Google Scholar]
  2. Engwall E. Enzyme immunoassay ELISA and EMIT. In: van Vankis H, Langone JJ, editors. Methods in Enzymology. New York: Academic Press; 1980. pp. 419–439. [DOI] [PubMed] [Google Scholar]
  3. Fourier A. Quantitative data on the Bombyx mori L. silkworm. Biochimie. 1979;61:283–320. doi: 10.1016/s0300-9084(79)80073-5. [DOI] [PubMed] [Google Scholar]
  4. Guilbert LI, Iscov NN. Partial replacement of serum by selenite, transferrin, albumin and lecithin in haemapoietic cell cultures. Nature. 1976;263:594–595. doi: 10.1038/263594a0. [DOI] [PubMed] [Google Scholar]
  5. Kato N, Sato S, Yamanaka A, Yamada H, Fuwa N, Nomira M. Silk protein, sericin, inhibits lipid peroxidation and tyrosinase activity. Biosci Biotechnol Biochem. 1998;62:145–147. doi: 10.1271/bbb.62.145. [DOI] [PubMed] [Google Scholar]
  6. Kelley DS, Becked JE, Potter VR. Effect of insulin, dexamethasone, and glucagon on the amino acid transport ability of four rat hepatoma cell lines and rat hepatocytes in culture. Cancer Res. 1978;38:4591–4601. [PubMed] [Google Scholar]
  7. Makishima F, Terada S, Mikami T, Suzuki E. Interleukin-6 is antiproliferative to a mouse hybridoma cell line and promotive for its antibody productivity. Cytotechnology. 1992;10:15–23. doi: 10.1007/BF00376096. [DOI] [PubMed] [Google Scholar]
  8. Minoura N, Aiba S, Gotoh Y, Tsukada M, Imai Y. Attachment and growth of cultured fibroblast cells on silk protein materials. J Biomed Mater Res. 1995;29:1215–1221. doi: 10.1002/jbm.820291008. [DOI] [PubMed] [Google Scholar]
  9. Murakami H, Masui H, Sato GH, Sueoka N, Chow TP, Kono-Sueoka T. Growth of hybridoma cells in serum-free medium; ethanolamine is an essential component. Proc Natl Acad Sci USA. 1982;79:1158–1162. doi: 10.1073/pnas.79.4.1158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Murakami H. Serum-free cultivation of plasmacytomas and hybridomas. In: Barnes DW, Sirbasku DA, Sato GH, editors. Methods for Serum-free Culture of Neuronal and Lymphoid Cells. New York: Alan R. Liss Inc.; 1984. pp. 197–206. [Google Scholar]
  11. Sasaki M, Kato N, Watanabe H, Yamada H. Silk protein, sericin, suppresses colon carcinogenesis induced by 1,2-dimethylhydrazine in mice. Oncol Rep. 2000;7:1049–1052. doi: 10.3892/or.7.5.1049. [DOI] [PubMed] [Google Scholar]
  12. Sasaki M, Yamada H, Kato N. Consumption of silk protein, sericin elevates intestinal absorption of zinc, iron, magnesium and calcium in rats. Nutr Res. 2000;20:1505–1511. doi: 10.1016/S0271-5317(00)80031-7. [DOI] [Google Scholar]
  13. Tsujimoto K, Takagi H, Takahashi M, Yamada H, Nakamori S. Cryoprotective effect of the serine-rich repetitive sequence in silk protein sericin. J Biochem. 2001;129:979–986. doi: 10.1093/oxfordjournals.jbchem.a002946. [DOI] [PubMed] [Google Scholar]
  14. Voegeli R, Meier J, Blust R. Sericin silk protein, unique structure and properties. Cosmetics Toiletries. 1993;108:101–108. [Google Scholar]

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

RESOURCES