The roles of monocytes and T lymphocytes in the regulation of human peripheral blood erythroid burst-forming units (BFU-E) were studied in erythropoietin-containing plasma clot cultures of subpopulations of human blood mononuclear cells. BFU-E growth was decreased significantly after depletion of monocytes alone (mean 11% of expected, range 0 to 42% of expected) or depletion of both monocytes and T cells (mean 6.5% of expected, range 0.5 to 12% of expected) from mononuclear cells. T cell depletion did not impair BFU-E growth in vitro. Using 10(5) monocyte- and T lymphocyte-depleted mononuclear cells as target cells (less than 1% monocytes, less than 5% T cells), BFU-E growth was restored to 40% of expected by addition of 10(4) monocytes, and to 96% of expected by 10(5) monocytes alone. Addition of as many as 2 X 10(5) T cells but no monocytes resulted in stimulation to only 34% of expected BFU-E growth. Addition of 2 X 10(4) T cells, which alone did not affect BFU-E growth, could augment significantly the stimulatory effect of 5-20 X 10(3) monocytes on BFU-E growth. Thus, monocytes alone appear to be capable of stimulating BFU-E growth in vitro in the presence of erythropoietin. T cells also may make small quantities of BFU-E stimulators. However, it seems more likely that the most important role of T lymphocytes in BFU-E regulation in vitro is a result of interactions with monocytes and augmentation of monocyte production of stimulators of BFU-E growth.