In Vitro Assay for Erythropoietin: Erythroid Colony Formation in Methyl Cellulose Used for the Measurement of Erythropoietin in Plasma

Erythroid colony formation in methyl cellulose has been used for the measurement of erythropoietin in plasma. Livers from newborn mice less than 24 hr old were found to provide convenient target cells. Newborn mouse liver contains a substantial number of erythroid colony-forming cells (CFU-e) that have a high sensitivity to erythropoietin. the dose-response curve for erythropoietin reaching a plateau at 50 mU/mI. As little as 0.5 rn/mI of the hormone is detectable. Removal of cells that adhered to glass prior to culturing doubled the number of colonies formed in the presence of erythropoietin. Addition of untreated plasmas that showed high erythropoietin titers in the exhypoxic polycythemic mice assay gave variable results. Some of the plasmas stimulated colony formation actively and in a linear fashion. However. the majority of the plasmas were toxic to the cultures. Dialyzing the plasmas for 3 days against distilled water effectively removed the toxicity. Results obtamed with the method are in good agreement with the values found using the exhypoxic polycythemic mice assay.