Identity and Functional Properties of Novel Skin-Derived Fibroblast Lines (NS Series) That Support the Growth of Epidermal-Derived Dendritic Cell Lines

Abstract

We have established recently a series of unique cell lines (NS series) from dispase-separated mouse epidermis that promote the growth of epidermal-derived antigen-presenting cell lines (XS series). The purposes of this study were to determine the identity of NS cells and to characterize their functional properties. NS cells were distinguishable from leukocytes by the lack of typical surface markers and by the failure to respond to leukocyte growth factors. Despite their epidermal derivation, NS cells were distinct from keratinocytes by the absence of cytokeratins. On the other hand, NS cells were indistinguishable from lines of dermal fibroblasts by their a) morphology, b) surface phenotype, and c) intracellular deposits of type I collagen. Growth of the XS antigen-presenting lines has been promoted by co-culturing with gamma-irradiated NS cells, and this activity could be replaced with NS cell-conditioned media alone, but not with paraformaldehyde-fixed NS cells. Each clone derived from the NS01 line secreted XS cell-growth-promoting activity, and this activity was blocked by monoclonal antibodies against colony-stimulating factor-1 receptors. Dermal fibroblasts also promoted the growth of XS cells in a colony-stimulating factor-1-dependent manner. By contrast, culture supernatants from other cell lines derived from skin (e.g., Pam 212 keratinocytes, 7-17 dendritic epidermal T cells, or XS lines) failed to promote XS cell growth. These results indicate that NS cells belong to the fibroblast lineage and that they share the intrinsic property to secrete large amounts of colony-stimulating factor-1 with dermal fibroblasts. Dermal cells may support the growth of skin-associated antigen-presenting cells in vivo.