Epithelial-mesenchymal interactions enhance expression of collagen VII in vitro.
Expression of collagen VII, the major structural protein of the anchoring fibrils, was assessed in vitro using indirect immunofluorescence staining and immunoblotting of collagen VII isolated from cultures. A very low level of expression was observed in monocultures of normal human fibroblasts or keratinocytes, but the expression was greatly stimulated when fibroblasts and keratinocytes were co-cultured. Primary skin explants under culture conditions supporting growth of both cell types, or mixed co-cultures of purified fibroblasts and keratinocytes, exhibited clearly enhanced synthesis of collagen VII, and the intact tissue form of this collagen could be extracted from small co-cultures. Three-dimensional skin equivalents were constructed with fibroblasts embedded in a contracted gel of collagen I and III, with an overlying stratified keratinocyte epithelium. In these equivalents, expression of collagen VII was observed primarily in the lowest epithelial cells, indicating that these cells are the main manufacturers of collagen VII. Laminin and collagen IV were deposited in a linear fashion onto the epithelial-mesenchymal interface. The results suggest that epithelial-mesenchymal interactions, either through physical interactions and/or through soluble mediators, are necessary for efficient synthesis of collagen VII and biogenesis of the anchoring fibrils.
König
A
A
Department of Dermatology, University Hospital Zurich, Switzerland.
Bruckner-Tuderman
L
L
D002450:Cell Communication / Q000502:physiology; D002478:Cells, Cultured; D003094:Collagen / Q000032:analysis*; D046508:Culture Techniques; D004847:Epithelial Cells; D004848:Epithelium / Q000502:physiology; D005347:Fibroblasts / Q000502:physiology; D005455:Fluorescent Antibody Technique; D006801:Humans; D015603:Keratinocytes / Q000502:physiology; D012867:Skin / Q000737:chemistry* / Q000166:cytology
D003094:Collagen