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Blood, 1 March 2005, Vol. 105, No. 5, pp. 1937-1945.
Prepublished online as a Blood First Edition Paper on November 2, 2004; DOI 10.1182/blood-2004-09-3459.
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HEMATOPOIESIS
Prospective isolation and global gene expression analysis of the erythrocyte colony-forming unit (CFU-E)
Grzegorz Terszowski,
Claudia Waskow,
Peter Conradt,
Dido Lenze,
Jessica Koenigsmann,
Dirk Carstanjen,
Ivan Horak, and
Hans-Reimer Rodewald
From the Department for Immunology, University of Ulm, Ulm, Germany; Institute of Pathology, Charité, Benjamin Franklin University Hospital, Berlin, Germany; and Department of Molecular Genetics, Institute of Molecular Pharmacology, Berlin, Germany.
The erythrocyte colony-forming unit (CFU-E) is a rare bone marrow (BM) progenitor that generates erythrocyte colonies in 48 hours. The existence of CFU-Es is based on these colonies, but CFU-Es have not been purified prospectively by phenotype. We have separated the "nonstem," "nonlymphoid" compartment (lineage marker [lin]–c-Kit+Sca-1–IL-7R –) into interleukin 3 receptor negative (IL-3R–) and IL-3R+ subsets. Within IL-3R– but not IL-3R+ cells we have identified TER119–CD41–CD71+ erythrocyte-committed progenitors (EPs). EPs generate CFU-E colonies at about 70% efficiency and generate reticulocytes in vivo. Depletion of EPs from BM strongly reduces CFU-E frequencies. EPs lack potential for erythrocyte burst-forming unit, megakaryocyte, granulocyte (G), and monocyte (M) colonies, and for spleen colony-forming units. Chronically suppressed erythropoiesis in interferon consensus sequence-binding protein (ICSBP)–deficient BM is associated with reduced frequencies of both the EP population and CFU-E colonies. During phenylhydrazine-induced acute anemia, numbers of both the EP population and CFU-E colonies increase. Collectively, EPs (lin–c-Kit+Sca-1–IL-7R–IL-3R–CD41–CD71+) account for most, if not all, CFU-E activity in BM. As a first molecular characterization, we have compared global gene expression in EPs and nonerythroid GM progenitors. These analyses define an erythroid progenitor-specific gene expression pattern. The prospective isolation of EPs is an important step to analyze physiologic and pathologic erythropoiesis.
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