Membrane-associated chondroitin sulfate proteoglycan and fibronectin mediate the binding of hemopoietic progenitor cells to stromal cells

Abstract

The initial step in hemopoiesis is the binding of progenitor cells to stroma. What mediates this binding at the molecular level is not entirely clear. We have previously reported that the cell line FDCP-1, a factor-dependent hemopoietic progenitor cell, actively synthesizes a membrane-associated chondroitin sulfate (CS) proteoglycan (MA-PG) which is unstable. After the binding of the progenitor cell to stromal, the stability of the MA-PG is enhanced, suggesting its involvement in the binding of progenitor cells to the stroma. Since stromal cells possess pericellular fibronectin (FN), we examined the possibility that binding to stromal cells may involve interactions between MA-PG of FDCP-1 on the one side and pericellular FN in stromal cells on the other side. To examine this hypothesis, we developed a cell adherence assay to measure the binding of FDCP-1 cells to a monolayer of stromal cells or to FN-coated dishes. Cell binding was inhibited by a monoclonal antibody against CS as well as by free CS and heparin, suggesting the involvement of MA-PG in the binding. Pretreatment of FDCP-1 cells with chondroitinase ABC, which selectively removes the CS portion of the MA-PG, also affects binding to the stromal cells. The binding was also inhibited by a pentapeptide (GRGDS) which competes with the cell-binding domain of FN as well as by a monoclonal antibody anti-FN. We conclude that interactions between MA-PG and a putative integrin-like molecule in FDCP-1 and the heparin and the cell binding domains in pericellular FN in the stromal cells contribute to the stabilization of progenitor-stromal cell binding which originally comes about by homing receptors of progenitor cells.