Murine hematopoietic blast colony-forming cells and their progeny have distinctive membrane marker profiles

Two distinct bone marrow-derived blast colony-forming cells can generate colonies of lineage-restricted progenitor cells in agar cultures of murine bone marrow. Both cell types selectively had a Kit⁺ ScaI⁺ phenotype distinguishing them from most lineage-restricted progenitor cells. Multicentric blas...

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Bibliographic Details
Published in:	Proceedings of the National Academy of Sciences - PNAS Vol. 106; no. 45; pp. 19102 - 19107
Main Authors:	Metcalf, Donald, Ng, Ashley P, Loughran, Stephen J.L, Phipson, Belinda, Smyth, Gordon K, Di Rago, Ladina, Mifsud, Sandra
Format:	Journal Article
Language:	English
Published:	United States National Academy of Sciences 10-11-2009 National Acad Sciences
Subjects:	Animals B lymphocytes Biological Sciences Blasts Bone marrow Bone marrow cells Cell Culture Techniques - methods Cell Lineage Cell Membrane - metabolism Cells Cultured cells Cytokines Deoxyribonucleases, Type II Site-Specific - metabolism Flow Cytometry Hematopoietic stem cells Hematopoietic Stem Cells - cytology Hematopoietic Stem Cells - metabolism Interleukin-6 Ligands Lymphocytes Membranes Mice Mice, Inbred C57BL Molecules Progenitor cells Proto-Oncogene Proteins c-kit - metabolism Stem Cell Factor Stem cells
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Summary:	Two distinct bone marrow-derived blast colony-forming cells can generate colonies of lineage-restricted progenitor cells in agar cultures of murine bone marrow. Both cell types selectively had a Kit⁺ ScaI⁺ phenotype distinguishing them from most lineage-restricted progenitor cells. Multicentric blast colony-forming cells stimulated by stem cell factor plus interleukin-6 (IL-6) (BL-CFC-S) were separable from most dispersed blast colony-forming cells stimulated by Flt3 ligand and IL-6 (BL-CFC-F) using CD34 and Flt3R probes. Multicentric BL-CFC-S cofractionated with colony-forming units, spleen (CFU-S) supporting the possibility that the 2 cells may be identical. The colony populations generated by BL-CFC-S were similar in their phenotype and proliferative capacity to progenitor cells in whole bone marrow but the progeny of BL-CFC-F were skewed with an abnormally high proportion of Kit⁻ Flt3R⁺ cells whose clonogenic cells tended to generate only macrophage progeny. Both blast colony populations had a high percentage of GR1⁺ and Mac1⁺ cells but BL-CFC-F colonies also contained a significant population of B220⁺ and IL-7R⁺ cells relevant to the superior ability of BL-CFC-F colony cells to generate B lymphocytes and the known dependency of this process on Flt3 ligand and IL-7. The commitment events and phenotypic changes during the generation of differing progenitor cells in blast colonies can now be clonally analyzed in a convenient in vitro culture system.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Contributed by Donald Metcalf, September 15, 2009 Author contributions: D.M., A.P.N., and S.J.L.L. designed research; D.M., A.P.N., S.J.L.L., L.D.R., and S.M. performed research; D.M., A.P.N., S.J.L.L., B.P., and G.K.S. analyzed data; and D.M. wrote the paper.
ISSN:	0027-8424 1091-6490
DOI:	10.1073/pnas.0910354106