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Identification and Analysis of Mouse Erythroid Progenitors using the CD71/TER119 Flow-cytometric Assay
Published on: August 5, 2011
Fetal and maternal progenitor cells in co-culture respond equally to erythropoietin
R M Bohmer1, K L Johnson, D W Bianchi
1Division of Genetics, Department of Pediatrics, Tufts University School of Medicine, Boston, MA, USA. rbohmer@lifespan.org
Abstract:
To explore potentially selective growth conditions for fetal cells in cultures from the blood of pregnant women, we investigated if fetal and adult erythroid progenitors with different hemoglobin expression programs are differentially responsive to erythropoietin (EPO). Co-cultures of clonogenic cells from 12-week fetal and adult peripheral blood were established, and the development of erythropoietic cells was monitored using flow cytometric profiles of correlated cellular contents of fetal and adult hemoglobin (HbF and HbA, respectively). Adult nucleated red cells were classified as F+A-, F+A+ or F-A+. All fetal cells were F+A-. The population of F+A- cells was flow-sorted and fetal cells were identified by fluorescence in situ hybridization (FISH) using chromosome-specific probes. Delayed EPO addition revealed that all types of erythroid cells entered the EPO-dependent phase with similar kinetics, beginning at about Day 4. The data suggest that fetal and adult erythroid stem/progenitor cells have the same initial maturation kinetics in culture independent of their hemoglobin chain expression program. Fetal and adult cells with different hemoglobin profiles also showed similar EPO dose-response curves, determined for different intervals during the first 2 weeks of culture. Thus, the kinetics of entry into the phase of EPO dependence, as well as the sensitivity to EPO at various stages of development, are essentially the same for erythropoietic progenitor cells derived from adult and early fetal blood, which rules out the possibility of using the timing or concentration of EPO for the selective growth of fetal cells from the blood of pregnant women.
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