Integration of Langerhans-like cells into a human skin equivalent

Arch Dermatol Res. 2011 Mar;303(2):135-9. doi: 10.1007/s00403-010-1092-x. Epub 2010 Nov 11.

Abstract

Studies regarding cellular interactions between Langerhans cells and other skin cells are somehow hampered by the difficult cultivation of these cells in vitro. Here, we show that the human MUTZ-3 cell line can be differentiated into Langerhans-like cells in the presence of a cytokine cocktail including GM-CSF, TGF-β1 and TNF-α. We used the expression of langerin, CD1a, CCR6 and the intracellular presence of Birbeck granules to identify the differentiated MUTZ-3 cells (MUTZ-3-LCs). The aim of this study was to integrate MUTZ-3-LCs into a three-dimensional full-thickness skin model. On top of fibroblast-containing collagen matrix a mixture of primary human keratinocytes and MUTZ-3-LCs were seeded and cultured for 24 h. Subsequently, the models were lifted up to the air-liquid interface. Histological evaluation featured a fully stratified epidermis with all characteristic epidermal strata. Langerin-positive cells were detected suprabasally within the epidermis indicating that keratinocytes provide environmental conditions for long-time maintenance of MUTZ-3-LCs. These skin models provide a tool to further investigate the interactions between Langerhans-like cells and other skin cells and particularly learn more about the cutaneous immune response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Cell Differentiation
  • Cell Line
  • Cells, Cultured
  • Collagen
  • Cytokines / pharmacology
  • Flow Cytometry
  • Humans
  • Keratinocytes / physiology*
  • Langerhans Cells / cytology
  • Langerhans Cells / physiology*
  • Male
  • Microscopy, Electron, Transmission
  • Skin, Artificial*

Substances

  • Cytokines
  • Collagen