C onditioned media were prepared from human peripheral blood monocytes and human umbilical vein endothelial cells. These media were assayed for erythroid burst-promoting activity (BPA) using human peripheral blood monocyte-depleted mononuclear cells as targets and assessing the stimulatory effect of the conditioned media on growth of early erythroid progenitor cells. Both monocytes and endothelial cells produced modest amounts of detectable BPA. Addition of varying concentrations of media conditioned by monocytes to plateau concentrations (5-10%) of media conditioned by endothelial cells had no additive effect. Endothelial cells incubated in the presence of 50% monocyte-conditioned medium produced 2.5- to 6.6-fold more BPA than did endothelial cells incubated only in control tissue culture medium. In contrast, endothelial cell conditioned medium did not stimulate increased BPA production by monocytes. Neither neutrophil- nor marrow fibroblastoid cell-conditioned medium stimulated BPA production by endothelial cells. Therefore, both monocytes and endothelial cells produce BPA. Moreover, monocytes produce a monokine that, in turn, stimulates the production of BPA by endothelial cells. Inasmuch as a monokine also has been shown to stimulate production of granulocyte-macrophage colony-stimulating activity, we propose that monocytes play a critical role in regulating the production of humoral regulators of the very early stages of hemopoietic cell differentiation.