Skip to main content

Advertisement

SpringerLink
Log in

Human adult epidermal melanocytes cultured without chemical mitogens express the EGF receptor and respond to EGF

  • Original Paper
  • Published:
Archives of Dermatological Research Aims and scope Submit manuscript

Abstract

We describe a novel chemical mitogen-free in vitro culture technique for obtaining pure melanocyte cultures using normal human adult epidermis as a source. The culture medium consists equal parts of the commercially available Keratinocyte Basal and AIM-V media (both from Gibco), as basal medium, which is supplemented with fetal bovine serum, bovine pituitary extract and recombinant human epidermal growth factor (EGF). Melanocytes harvested from human adult skin proliferate extensively and can be passaged serially up to 10–15 times using this medium. We have verified the identity of the cultured cells by tyrosinase mRNA expression and TRP-1 protein staining. Moreover, we showed that autologous human serum alone, without additional supplements is able to provide sufficient growth support for the cultured cells in the basal medium, making this culture technique suitable for autologous melanocyte transplantation. In this culture system normal human adult melanocytes expressed both EGF receptor (EGFR) mRNA and protein and EGF showed a dose dependent mitogenic effect on the cells. EGF itself had no significant influence on EGFR mRNA expression.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Albino AP, Vidal MJ, McNutt NS, Shea CR, Prieto VG, Nanus DM, Palmer JM, Hayward NK (1994) Mutation and expression of the p53 gene in human malignant melanoma. Melanoma Res 4:35–45

    Article  PubMed  CAS  Google Scholar 

  2. Bardeesy N, Kim M, Xu J, Kim RS, Shen Q, Bosenberg MW, Wong WH, Chin L (2005) Role of epidermal growth factor receptor signaling in RAS-driven melanoma. Mol Cell Biol 25:4176–4188

    Article  PubMed  CAS  Google Scholar 

  3. Clark AJ, Ishii S, Richert N, Merlino GT, Pastan I (1985) Epidermal growth factor regulates the expression of its own receptor. Proc Natl Acad Sci USA 82:8374–8378

    Article  PubMed  CAS  Google Scholar 

  4. Donatien P, Surleve-Bazeille JE, Thody AJ, Taieb A (1993) Growth and differentiation of normal human melanocytes in a TPA-free, cholera toxin-free, low-serum medium and influence of keratinocytes. Arch Dermatol Res 285:385–392

    Article  PubMed  CAS  Google Scholar 

  5. Eisinger M, Marko O (1982) Selective proliferation of normal human melanocytes in vitro in the presence of phorbol ester and cholera toxin. Proc Natl Acad Sci USA 79:2018–2022

    Article  PubMed  CAS  Google Scholar 

  6. Eisinger M, Marko O, Ogata S, Old LJ (1985) Growth regulation of human melanocytes: mitogenic factors in extracts of melanoma, astrocytoma, and fibroblast cell lines. Science 229:984–986

    Article  PubMed  CAS  Google Scholar 

  7. Elder DE, Rodeck U, Thurin J, Cardillo F, Clark WH, Stewart R, Herlyn M (1989) Antigenic profile of tumor progression stages in human melanocytic nevi and melanomas. Cancer Res 49:5091–5096

    PubMed  CAS  Google Scholar 

  8. Ellis DL, King LE Jr, Nanney LB (1992) Increased epidermal growth factor receptors in melanocytic lesions. J Am Acad Dermatol 27:539–546

    PubMed  CAS  Google Scholar 

  9. Falabella R, Escobar C, Borrero I (1989) Transplantation of in vitro-cultured epidermis bearing melanocytes for repigmenting vitiligo. J Am Acad Dermatol 21:257–264

    Article  PubMed  CAS  Google Scholar 

  10. Gilchrest BA, Vrabel MA, Flynn E, Szabo G (1984) Selective cultivation of human melanocytes from newborn and adult epidermis. J Invest Dermatol 83:370–376

    Article  PubMed  CAS  Google Scholar 

  11. Gordon-Thomson C, Mason RS, Moore GP (2001) Regulation of epidermal growth factor receptor expression in human melanocytes. Exp Dermatol 10:321–328

    Article  PubMed  CAS  Google Scholar 

  12. Grahn JC, Isseroff RR (2004) Human melanocytes do not express EGF receptors. J Invest Dermatol 123:244–246

    Article  PubMed  CAS  Google Scholar 

  13. Halaban R (2000) The regulation of normal melanocyte proliferation. Pigment Cell Res 13:4–14

    Article  PubMed  CAS  Google Scholar 

  14. Halaban R, Ghosh S, Baird A (1987) bFGF is the putative natural growth factor for human melanocytes. In Vitro Cell Dev Biol 23:47–52

    Article  PubMed  CAS  Google Scholar 

  15. Halaban R, Pomerantz SH, Marshall S, Lambert DT, Lerner AB (1983) Regulation of tyrosinase in human melanocytes grown in culture. J Cell Biol 97:480–488

    Article  PubMed  CAS  Google Scholar 

  16. Hara M, Yaar M, Gilchrest BA (1995) Endothelin-1 of keratinocyte origin is a mediator of melanocyte dendricity. J Invest Dermatol 105:744–748

    Article  PubMed  CAS  Google Scholar 

  17. Hu F, Staricco RJ, Pinkus H, Fosnaugh RP (1957) Human melanocytes in tissue culture. J Invest Dermatol 28:15–32

    PubMed  CAS  Google Scholar 

  18. Kitano Y, Okada N (1983) Separation of the epidermal sheet by dispase. Br J Dermatol 108:555–560

    Article  PubMed  CAS  Google Scholar 

  19. Lahav R (2005) Endothelin receptor B is required for the expansion of melanocyte precursors and malignant melanoma. Int J Dev Biol 49:173–180

    Article  PubMed  CAS  Google Scholar 

  20. Lazar-Molnar E, Hegyesi H, Toth S, Falus A (2000) Autocrine and paracrine regulation by cytokines and growth factors in melanoma. Cytokine 12:547–554

    Article  PubMed  CAS  Google Scholar 

  21. Lerner AB, Halaban R, Klaus SN, Moellmann GE (1987) Transplantation of human melanocytes. J Invest Dermatol 89:219–224

    Article  PubMed  CAS  Google Scholar 

  22. Lontz W, Olsson MJ, Moellmann G, Lerner AB (1994) Pigment cell transplantation for treatment of vitiligo: a progress report. J Am Acad Dermatol 30:591–597

    Article  PubMed  CAS  Google Scholar 

  23. Mattei S, Colombo MP, Melani C, Silvani A, Parmiani G, Herlyn M (1994) Expression of cytokine/growth factors and their receptors in human melanoma and melanocytes. Int J Cancer 56:853–857

    Article  PubMed  CAS  Google Scholar 

  24. Mirmohammadsadegh A, Hassan M, Gustrau A, Doroudi R, Schmittner N, Nambiar S, Tannapfel A, Ruzicka T, Hengge UR (2005) Constitutive expression of epidermal growth factor receptors on normal human melanocytes. J Invest Dermatol 125:392–394

    PubMed  CAS  Google Scholar 

  25. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63

    Article  PubMed  CAS  Google Scholar 

  26. Nielsen HI, Don P (1984) Culture of normal adult human melanocytes. Br J Dermatol 110:569–580

    Article  PubMed  CAS  Google Scholar 

  27. Olsson MJ, Juhlin L (1993) Repigmentation of vitiligo by transplantation of cultured autologous melanocytes. Acta Derm Venereol 73:49–51

    PubMed  CAS  Google Scholar 

  28. Olsson MJ, Juhlin L (1995) Transplantation of melanocytes in vitiligo. Br J Dermatol 132:587–591

    Article  PubMed  CAS  Google Scholar 

  29. Rodeck U (1993) Growth factor independence and growth regulatory pathways in human melanoma development. Cancer Metastasis Rev 12:219–226

    Article  PubMed  CAS  Google Scholar 

  30. Shahbazi M, Pravica V, Nasreen N, Fakhoury H, Fryer AA, Strange RC, Hutchinson PE, Osborne JE, Lear JT, Smith AG, Hutchinson IV (2002) Association between functional polymorphism in EGF gene and malignant melanoma. Lancet 359:397–401

    Article  PubMed  CAS  Google Scholar 

  31. Shibutani M, Lazarovici P, Johnson AC, Katagiri Y, Guroff G (1998) Transcriptional down-regulation of epidermal growth factor receptors by nerve growth factor treatment of PC12 cells. J Biol Chem 273:6878–6884

    Article  PubMed  CAS  Google Scholar 

  32. Swope VB, Medrano EE, Smalara D, Abdel-Malek ZA (1995) Long-term proliferation of human melanocytes is supported by the physiologic mitogens alpha-melanotropin, endothelin-1, and basic fibroblast growth factor. Exp Cell Res 217:453–459

    Article  PubMed  CAS  Google Scholar 

  33. Yaar M, Gilchrest BA (2004) Melanocyte biology: before, during, and after the Fitzpatrick era. J Invest Dermatol 122:xxvii-xxxix

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grants ETT 027/2001, GVOP-3.2.2.-2004-07-0010/3.0, OTKA K61541, NI 62007. Márta Széll was supported by the Bolyai Foundation of the Hungarian Academy of Sciences. Zsuzsanna Bata-Csörgő is a recipient of the Széchenyi István Fellowship of the Hungarian Ministry of Education.

Author information

Authors and Affiliations

  1. Faculty of Medicine, Department of Dermatology and Allergology, University of Szeged, 6720, Szeged, Korányi fasor 6, Hungary

    Gábor Szabad, Bernadett Kormos, Anna Kenderessy Szabó, Attila Dobozy, Lajos Kemény & Zsuzsanna Bata-Csörgő

  2. Dermatological Research Group of the Hungarian Academy of Sciences, Szeged, Hungary

    Andor Pivarcsi, Márta Széll, Attila Dobozy, Lajos Kemény & Zsuzsanna Bata-Csörgő

  3. Faculty of Pharmacy, Department of Clinical Pharmacy, University of Szeged, Szeged, Hungary

    Kornélia Kis

Authors
  1. Gábor Szabad
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bernadett Kormos
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andor Pivarcsi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Márta Széll
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kornélia Kis
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Anna Kenderessy Szabó
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Attila Dobozy
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Lajos Kemény
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Zsuzsanna Bata-Csörgő
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bernadett Kormos.

Additional information

Gábor Szabad and Bernadett Kormos contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Szabad, G., Kormos, B., Pivarcsi, A. et al. Human adult epidermal melanocytes cultured without chemical mitogens express the EGF receptor and respond to EGF. Arch Dermatol Res 299, 191–200 (2007). https://doi.org/10.1007/s00403-007-0742-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00403-007-0742-0

Keywords

Search

Navigation