INVITRO ASSAY FOR ERYTHROPOIETIN - ERYTHROID COLONY FORMATION IN METHYL CELLULOSE USED FOR THE MEASUREMENT OF ERYTHROPOIETIN IN PLASMA

  • 1 January 1979
    • journal article
    • research article
    • Vol. 53  (6) , 1172-1181
Abstract
Erythroid colony formation in methyl cellulose was used for the measurement of erythropoietin in plasma. Livers from newborn mice less than 24 h old provided convenient target cells. Newborn mouse liver contains a substantial number of erythroid colony-forming cells (CFU-e) that had a high sensitivity to erythropoietin, the dose-response curve for erythropoietin reaching a plateau at 50 mU[units]/ml. As little as 0.5 m/ml of the hormone was 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. The majority of the plasmas were toxic to the cultures. Dialyzing the plasmas for 3 days against distilled water effectively removed the toxicity. Results obtained with the method were in good agreement with the values found using the exhypoxic polycythemic mice assay.