Fibronectin Mediates Cytokinesis and Growth of Rat Follicular Cells in Serum-free Medium

In this study, the growth of rat follicular (RF-1) cells was severely depressed when the cells were subcultured by trypsinization directly into serum-free medium supplemented with insulin, transferrin and hydrocortisone, which are required for growth of these cells in vitro. Within 24 hr after plating, 50-65% of the cells became binucleated, indicating lack of cytokinesis. However, the addition of human plasma fibronectin (8 microgram/ml) to the serum-free medium eliminated cell binucleation and enhanced cell growth considerably. Fibronectin had the same effect when RF-1 cells were plated into tissue culture dishes on which fibronectin had been absorbed, and cells were inoculated into fibronectin-free medium. Cell binucleation and poor growth in serum-free medium occurred when the cells were subcultured by trypsinization, EGTA treatment or detachment of mitotic cells. Under some conditions, cells could be "rescued" if fibronectin was added soon after inoculation, indicating that fibronectin was needed mainly during a limited time when the subcultured cells were attaching to the tissue culture substratum. Our findings describe an adhesive activity for fibronectin which circumvents the serum preincubation usually needed after RF-1 cells are subcultured for growth in serum-free medium. They also indicate the importance of fibronectin-mediated adhesion for cytokinesis processes of these cells.