ERYTHROPOIETIN BIOASSAYS USING FETAL MOUSE-LIVER CELLS - VALIDATIONS AND TECHNICAL IMPROVEMENTS

  • 1 January 1983
    • journal article
    • research article
    • Vol. 11  (7) , 590-600
Abstract
An erythropoietin (Ep) bioassay using the uptake of 125I-deoxyuridine (125I-UdR) into whole cells cultured in micro-titer plates was compared with an established method where the cells were cultured in 1 ml volumes using the incorporation of 59Fe into heme as the endpoint. The validity of the substitution of 125I-UdR uptake into whole cells as a replacement for 59Fe incorporation into heme as an assay endpoint and the adaptation of the method to a semi-micro scale with automated cell harvesting was demonstrated. A culture time significantly shorter than 24 h is demonstrated not to be of practical benefit. Two methods are proposed to saturate the growth promoting effects of Fe containing transferrin. The performance of a semi-micro, serum-containing bioassay employing untreated serum as the test material is shown to be superior to previous systems for the biological measurement of Ep.