title: Directed differentiation and mass cultivation of pure erythroid progenitors from mouse embryonic stem cells.
abstract: 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.
authors.affiliation: Research Institute of Molecular Pathology, Vienna Biocenter, Dr Bohr Gasse 7, 1030 Vienna, Austria. carotta@imp.univie.ac.at
authors.forename: Sebastian
authors.initials: S
authors.lastname: Carotta
authors.forename: Sandra
authors.initials: S
authors.lastname: Pilat
authors.forename: Andreas
authors.initials: A
authors.lastname: Mairhofer
authors.forename: Uwe
authors.initials: U
authors.lastname: Schmidt
authors.forename: Helmut
authors.initials: H
authors.lastname: Dolznig
authors.forename: Peter
authors.initials: P
authors.lastname: Steinlein
authors.forename: Hartmut
authors.initials: H
authors.lastname: Beug
mesh_terms: D000375:Aging / Q000502:physiology; D000818:Animals; D001853:Bone Marrow / Q000473:pathology; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D004900:Erythroblasts / Q000166:cytology*; D004912:Erythrocytes / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D006454:Hemoglobins / Q000378:metabolism; D008099:Liver / Q000166:cytology; D051379:Mice; D018345:Mice, Knockout; D009369:Neoplasms / Q000097:blood / Q000473:pathology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000172:deficiency / Q000235:genetics
chemical_list: D006454:Hemoglobins; D040301:Vascular Endothelial Growth Factor Receptor-2