Fibronectin-mediated keratinocyte migration and initiation of fibronectin receptor function in vitro

Abstract

Cell suspensions of human keratinocytes, freshly isolated from skin specimens, did not express plasma fibronectin (pFN) receptor function in short-term assays for cell attachment and spreading on pFN-coated culture dishes and binding and phagocytosis of pFN-coated latex beads. These activities were expressed, however, by the cells harvested from primary keratinocyte cultures after 2-4 days of culture. Analysis of the cell types arising during primary cultures, based on staining with antikeratin antibodies and bullous pemphigoid (BP) serum, revealed that about 90% of the originally isolated cell population consisted of keratinocytes (keratin-positive) and 30% were basal cells (BP antigen-positive). After 2 days of culture, 95% of the cells were keratinocytes and 70% were basal cells. In vitro initiation of pFN receptor function also was observed in cells harvested from epidermal explants. After 9 days in culture, the cells that migrated out of the explants also were active in short-term cell adhesion assays, while cells remaining in the central region of the explant had much less activity. In related studies, the role of pFN in epidermal cell migration was analyzed, and it was found that anti-pFN antibodies inhibited migration of keratinocytes out of epidermal explants. Addition of preimmune IgG, however, had no effect. It appears, therefore, that pFN is important in all aspects of keratinocyte adhesion, and the expression of pFN receptor function may be a critical activation step necessary for basal cell phagocytosis and migration during wound healing.