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Blood, 15 September 2004, Vol. 104, No. 6, pp. 1873-1880.
Prepublished online as a Blood First Edition Paper on May 27, 2004; DOI 10.1182/blood-2004-02-0570.
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RED CELLS
Directed differentiation and mass cultivation of pure erythroid progenitors from mouse embryonic stem cells
Sebastian Carotta,
Sandra Pilat,
Andreas Mairhofer,
Uwe Schmidt,
Helmut Dolznig,
Peter Steinlein, and
Hartmut Beug
From the Research Institute of Molecular Pathology (IMP), Vienna Biocenter; the Institute of Immunology, Vienna International Research Cooperation Center (VIRCC); and the Institute of Ultrastructural Pathology, General Hospital Vienna (AKH), Vienna, Austria.
Differentiating embryonic stem (ES) cells are an increasingly important source of hematopoietic progenitors, useful for both basic research and clinical applications. Besides their characterization in colony assays, protocols exist for the cultivation of lymphoid, myeloid, and erythroid cells. With the possible exception of mast cells, however, long-term expansion of pure hematopoietic progenitors from ES cells has not been possible without immortalization caused by overexpression of exogenous genes. Here, we describe for the first time an efficient yet easy strategy to generate mass cultures of pure, immature erythroid progenitors from mouse ES cells (ES-EPs), using serum-free medium plus recombinant cytokines and hormones. ES-EPs represent long-lived, adult, definitive erythroid progenitors that resemble immature erythroid cells expanding in vivo during stress erythropoiesis. When exposed to terminal differentiation conditions, ES-EPs differentiated into mature, enucleated erythrocytes. Importantly, ES-EPs injected into mice did not exhibit tumorigenic potential but differentiated into normal erythrocytes. Both the virtually unlimited supply of cells and the defined culture conditions render our system a valuable tool for the analysis of factors influencing proliferation and maturation of erythroid progenitors. In addition, the system allows detailed characterization of processes during erythroid proliferation and differentiation using wild-type (wt) and genetically modified ES cells.
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