Molecular Biology Reports

, Volume 6, Issue 3, pp 153–158 | Cite as

Protein biosynthesis in cultured human hair follicle cells

  • P. J. J. M. Weterings
  • A. J. M. Vermorken
  • H. Bloemendal
Article

Abstract

A new technique has been used for culturing human keratinocytes. The cells grow on the basement membrane-like capsules of bovine lenses. Lens cells were removed from the capsules by rigid trypsinization. In order to exclude any contamination with remaining living cells the isolated capsules were irradiated with X-rays at a dose of 10,000 rad. In this way human epithelial cells can be brought in culture from individual hair follicles. Since feeder cells are not used in this culture technique, the biosynthesis of keratinocyte proteins can be studied in these cultures. The newly synthesized proteins can be separated into a water-soluble, a urea-soluble, and a urea-insoluble fraction. Product analysis has been performed on the first two fractions revealing protein patterns identical to those of intact hair follicles. Product analysis of the urea-soluble fractions of microdissected hair follicles shows that the protein pattern of the cultured keratinocytes resembles the protein pattern of the hair follicle sheath. Studies on the metabolism of benzo(a)pyrene revealed that the enzyme aryl hydrocarbon hydroxylase (AHH) is present in cultured hair follicle cells. A possible use of our culture system for eventual detection of inherited predisposition for smoking-dependent lung cancer is discussed.

Keywords

Pyrene Hair Follicle Human Hair Protein Pattern Human Epithelial Cell 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    GrimmI., T.F.Wienker & H.H.Ropers. Fabry's disease: heterozygote detection by hair root analysis. Hum. Genet. 32, 329–334 (1978).Google Scholar
  2. 2.
    VermorkenA.J.M., P.J.J.M.Weterings, G.Th.Spierenburg, C.A.vanBennekom, P.Wirtz, C.H.M.M.deBruyn, & T.L.Oei. Fabry's disease: biochemical and histochemical studies on hair roots for carrier detection. Br. J. Dermatology 98, 191–196. (1978).Google Scholar
  3. 3.
    Vermorken, A.J.M., C.A. van Bennekom, C.H.M.M. de Bruyn, T.L. Oei & J. Frohlich. Heterozygote detection in Fabry's disease using mailed hair roots. Br. J. Dermatology (in press).Google Scholar
  4. 4.
    GartlerS.M., T.C.Scott, J.L.Goldstein, B.Campbell & R.Sparkes. Lesch-Nyman syndrome: rapid detection of heterozygotes by use of hair follicles. Science 172, 572–574 (1971).Google Scholar
  5. 5.
    DeBruynC.H.M.M., T.L.Oei & B.G.A.terHaar. Studies on hair roots for carrier detection in hypoxanthineguanine phosphoribosyl transferase deficiency. Clin. Genet. 5, 449–456 (1974).Google Scholar
  6. 6.
    RomeoG., A.Rinaldi, F.Urbanus & G.Filippi. Hair root versus red cell individual phenotype in Sardinian heterozygotes for G6PD deficiency (Mediterranean type). Amer. J. Genet. 28, 506–513 (1976).Google Scholar
  7. 7.
    VermorkenA.J.M., P.Wirtz, G.Th.Spierenburg, C.A.vanBennekom, C.H.M.M.deBruyn & T.L.Oei. Glucose-6-phosphate dehydrogenase deficiency: Biochemical and histochemical studies on hair roots for carrier detection. J. Clin. Chem. Clin. Biochem. 17, 325–329 (1979).Google Scholar
  8. 8.
    VermorkenA.J.M., G.Th.Spierenburg, C.A.vanBennekom, C.H.M.M.deBruyn, T.L.Oei, G.E.J.Staal & R.A.Geerdink. Heterozygote detection in glucose-6-phosphate dehydrogenase deficiency: limitation of hair follicle analysis. Clin. Genet. 16, 353–356 (1979).Google Scholar
  9. 9.
    RheinwaldJ.G. & H.Green. Epidermal growth factor and the multiplication of cultured human epidermal keratinocytes. Nature 265, 421–424 (1977).Google Scholar
  10. 10.
    RheinwaldJ.G. & H.Green. Formation of a keratinizing epithelium in culture by a cloned cell line derived from a teratoma. Cell 6, 317–330 (1975).Google Scholar
  11. 11.
    RheinwaldJ.G. & H.Green. Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell 6, 331–334 (1975).Google Scholar
  12. 12.
    KondoS., K.Aso & M.Nauta. Culture of normal human epidermal cells with 3T3 feeders on millipore filters. J. Invest. Dermatology 72, 85–87 (1979).Google Scholar
  13. 13.
    Hukkelhoven, M.W.A.C., A.J.M. Vermorken & H. Bloemendal. A novel method for culturing epithelial cells on a biological substrate. Prep. Biochemistry 1980 (in press).Google Scholar
  14. 14.
    VermorkenA.J.M., P.J.J.M.Weterings & H.Bloemendal. Protein biosynthesis in isolated human scalp hair follicles. Molec. Biol. Rep. 4, 211–216 (1978).Google Scholar
  15. 15.
    LaemmliU.K. Cleavage of structural proteins during the assembly of the head of bacteriophage. T4. Nature 227, 680–685. (1970).Google Scholar
  16. 16.
    WeberK. & M.Osborn. The reliability of molecular weight determination by dodecyl sulphate polyacrylamide gel electrophoresis. J. Biol. Chem. 244, 4406–4412 (1969).Google Scholar
  17. 17.
    BonnerW.M. & R.A.Laskey. A film detection method for tritium labeled protein and nucleic acids in polyacrylamide gels. Eur. J. Biochem. 46, 83–88 (1974).Google Scholar
  18. 18.
    BernsA.J.M. & H.Bloemendal. Translation of mRNA from vertebrate eye lenses. In: Methods in Enzymology (pub. Academic Press, New York-London) Vol. XXX pp. 675–694 (1974).Google Scholar
  19. 19.
    VermorkenA.J.M., C.M.A.A.Goos, H.M.J.Roelofs, P.Th.Henderson & H.Bloemendal. Metabolism of benzo(a)pyrene in isolated human scalp hair follicles. Toxicology 14, 109–116 (1979).Google Scholar
  20. 20.
    KellermanG., M.Luyten-Kellerman & C.R.Shaw. Genetic variation of aryl hydrocarbon hydroxylase in human lymphocytes. Am. J. Hum. Genet. 25, 327–331 (1973).Google Scholar
  21. 21.
    ThorgeirssonS.S. & D.W.Nebert. The Ah locus and the metabolism of chemical carcinogens and other foreign components, in: Advances in cancer research, KleinF. and WeinhouseS. eds., Academic Press, New York, pp. 149–193 (1977).Google Scholar
  22. 22.
    Vermorken, A.J.M. & H. Bloemendal. Is the hair root a useful biopsy tissue for detection of the inherited predisposition for lung cancer? In: Tumour markers: impact and prospects. Elsevier/North-Holland Biochemical Press B.V., Boelsma, E. and Rumke, Ph. eds., pp. 305–324 (1979).Google Scholar
  23. 23.
    FoxC.H., K.Selkink, F.M.Price, R.G.Croy, U.K.Sanford & M.Cottler-Fox. Metabolism of benzo(a)-pyrene by human epidermal cells in vitro. Cancer research 35, 3351–3357 (1975).Google Scholar
  24. 24.
    Weterings, P.J.J.M., A.J.M. Vermorken & H. Bloemendal. A method for culturing human hair follicle cells. Br. J. Dermatology 1980 (in press).Google Scholar

Copyright information

© Dr. W. Junk B.V. Publishers 1980

Authors and Affiliations

  • P. J. J. M. Weterings
    • 1
  • A. J. M. Vermorken
    • 1
  • H. Bloemendal
    • 2
  1. 1.Research Unit for Cellular Differentiation and TransformationUniversity of NijmegenNijmegenThe Netherlands
  2. 2.Department of BiochemistryUniversity of NijmegenNijmegenThe Netherlands

Personalised recommendations