Organotypic Culture of Human Skin to Study Melanocyte Migration

I. CAROLINE LE POOLE, RENÉ M.J.G.J. VAN DEN WIJNGAARD, WIETE WESTERHOF, JAN A.M.A. DORMANS, FRANK M. VAN DEN BERG, RONALD P. VERKRUISEN, KOERT P. DINGEMANS, PRANAB K. DAS*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

An ex vivo model system was developed to investigate melanocyte migration. Within this model system, melanocytes migrate among other epidermal cells in the epibolic outgrowth of skin explants. This process is initiated by loss of contact inhibition of epidermal cells at the rim of the explants and by locally produced chemotactic factors. Punch biopsies provided explants of reproducible diameter. Optimal culture conditions include medium consisting of Dulbecco's Minimal Essential Medium containing 10% inactivated normal human serum and placement of explants epidermal side up at the air‐liquid interphase. Within 7 days, epidermal cells completely surround the explant. Approximately 3 days after the onset of keratinocyte migration, melanocytes distribute themselves within the newly formed epidermis. Throughout the 7‐day culture period, melanocytes and keratinocytes show maintenance of subcellular morphology, and the dermo‐epidermal junction remains intact. Melanocyte migration was quantified using immunoperoxidase staining in combination with light microscopy and computer‐aided image analysis. Preliminary results using the model system to compare migration in control and nonlesional vitiligo skin indicate that no inherent migration defect is responsible for impaired repigmentation of vitiligo lesions. The organotypic culture model system allows for investigations on melanocytes within their environment of autologous epidermal and dermal components, closely resembling in vivo circumstances in human skin.

Original languageEnglish (US)
Pages (from-to)33-43
Number of pages11
JournalPigment Cell Research
Volume7
Issue number1
DOIs
StatePublished - Jan 1 1994

Keywords

  • Chemoattractants
  • Epiboly
  • Explants
  • Model system
  • Repigmentation
  • Vitiligo

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science
  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

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