Blackwell Publishing Journal Backfiles 1879-2005
In order to provide a model for in vitro studies of the interactions between skin and the nervous system, we have performed a co-culture of epidermal cells and neurons. We used a tri-compartmented box with separated domains. In the central part of this box, we put an epidermal suspension (4 millions cells/ml) obtained from biopsies of human skin. Around this central domain, we disposed sensorial neurons from the dorsal root ganglia of rats. The periphery was occupied by neurons from the dorsal horn of spinal cord. Sensorial neurons grew in low density (500 cells), on a glial layer, in a medium conditioned by astrocytes. After 15 days of culture, cells were fixed and stained with monoclonal antibodies directed against PGP 9.5, keratins, or cytokeratin 20 (Merkel cells). We obtained a co-culture with three identifiable territories, equivalents of epidermis, root ganglia, and spinal cord. Nervous fibers specifically grew from the sensorial neurons to epidermal cells or to the spinal cord equivalent. We observed synapse-like contacts between nerve endings and Merkel cells or keratinocytes. This model allows us to reconstruct in vitro an equivalent of sensitive nerve fibers, connected for one part to a spinal cord equivalent and on the other part to an epidermis equivalent. Such a model could be used to understand the origin and the function of Merkel cells in the epidermis and to study synapses in the skin.
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