Cellular Mechanisms for Increased Fetal Hemoglobin Production in Culture: EVIDENCE FOR CONTINUOUS COMMITMENT TO FETAL HEMOGLOBIN PRODUCTION DURING BURST FORMATION

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Abstract

Using microscopic immunodiffusion assays and microdensitometric analysis of pericellular immunoprecipitate, the percentage of nucleated erythrocytes containing fetal hemoglobin (FNRBC) and the mean picograms of fetal or adult hemoglobin per nucleated erythrocyte (picograms HbF/NRBC, picograms HbA/NRBC) were assayed in 14-d-old colonies (bursts) derived from peripheral blood erythroid progenitors. In the peripheral blood of 11 normal adults only 2.2±0.5% (mean±SE) erythrocytes contained HbF whereas pooled bursts from the same subjects revealed a 13-fold increase in the percentage of FNRBC (29.6±3.9%). In culture both the picograms HbF/NRBC (5.2±0.4) and the picograms HbA/NRBC (27.7±1.5) are increased ∼20% above the mean in vivo levels in NRBC from normal bone marrow aspirates. Analysis of each of 58 bursts from one subject demonstrated that FNRBC are present in all bursts and range from 5.0 to 95.0% of the total NRBC per burst. The percent FNRBC in each burst was neither correlated with picograms HbF/NRBC per burst nor with picograms HbA/NRBC per burst. Individual subcolonies from one burst in each of two subjects demonstrated between 3 and 81% FNRBC.

Authors

George J. Dover, Makio Ogawa