Dual role of fibronectin in hematopoietic differentiation

The adhesive glycoprotein fibronectin provides anchorage for fibroblasts and hematopoietic progenitor cells in vitro. Fibronectin also demonstrates growth factor activity for fibroblasts; however, there is no available information regarding its role as a hematopoietic growth factor. To distinguish growth factor activity of fibronectin from its anchorage activity for hematopoietic progenitors, we assessed the ability of purified human plasma fibronectin to promote human bone marrow erythroid, granulocyte-macrophage (GM) and mixed granulocyte- erythroid-macrophage-megakaryocyte (GEMM) colony formation in liquid suspension, methylcellulose, and fibrin clots under serum-free conditions. Addition of fibronectin to methylcellulose cultures, or to cultures formed in fibrin clots, using fibrinogen depleted of fibronectin by preadsorption over gelatin-Sepharose and clotted with thrombin, resulted in up to a twofold enhancement of proliferation of erythroid burst-forming units (BFU-E), erythroid colony-forming units (CFU-E), and CFU-GEMM. This effect was concentration-dependent up to a fibronectin supplement of 100 micrograms/mL. By contrast, CFU-GM proliferation was not affected by the addition of fibronectin to the cultures. Fibronectin-adherent marrow cells overlaid with liquid medium formed both early and late-appearing erythroid colonies, whereas similarly cultured plastic-adherent marrow cells did not. Erythroid colony formation was observed in cultures of fibronectin-adherent marrow cells overlaid with methylcellulose but not in cultures of plastic-adherent marrow cells under the same conditions. Finally, the erythroid growth-promoting activity of fibronectin was inhibited by arginyl-glycyl-aspartyl-serine (RGDS), a tetrapeptide that competitively blocks the interaction of fibronectin with its receptor. We conclude that fibronectin plays a dual role in hematopoiesis: providing (a) anchorage for erythroid and primitive progenitors, and (b) as a proliferative stimulus for these hematopoietic cells. Both activities may be mediated by the cell adhesion domain of the molecule.