Mouse fetal liver tissue has been cultured and shown to produce and release into the culture medium an erythropoietically active substance for up to 30 days of culture. Since this substance can be completely neutralized by an antiserum to erythropoietin and shows a dose-- response relationship in the plethoric mouse assay, it is suggested that the culture medium contains erythropoietin, a hormone important in the regulation of erythropoiesis. Using this procedure, we have obtained the equivalent of about 20.7 unites of erythropoietin from five T-flasks (75 sq cm) over the 30-day culture period.