Prospective isolation and molecular characterization of hematopoietic stem cells with durable self-renewal potential

• Published in: Blood 2009-06, Vol.113 (25), p.6342-6350
• Main Authors: Kent, David G., Copley, Michael R., Benz, Claudia, Wöhrer, Stefan, Dykstra, Brad J., Ma, Elaine, Cheyne, John, Zhao, Yongjun, Bowie, Michelle B., Zhao, Yun, Gasparetto, Maura, Delaney, Allen, Smith, Clayton, Marra, Marco, Eaves, Connie J.
• Format: Article
• Language: English
• Subjects: Animals; Animals, Congenic; Antigens, CD - analysis; Antigens, Differentiation - analysis; Biological and medical sciences; Bone Marrow Cells - cytology; Cell Division; Cell Separation - methods; Cells, Cultured - transplantation; Flow Cytometry - methods; Gene Expression Profiling; Hematologic and hematopoietic diseases; Hematopoietic Stem Cell Transplantation; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - metabolism; Immunophenotyping; Leukocyte Common Antigens - analysis; Liver - cytology; Liver - embryology; Medical sciences; Mice; Mice, Inbred C57BL; Phospholipase D - analysis; Radiation Chimera; Receptors, Cell Surface - analysis; rhoB GTP-Binding Protein - analysis; rhoB GTP-Binding Protein - genetics; Signaling Lymphocytic Activation Molecule Family Member 1; von Willebrand Factor - analysis; von Willebrand Factor - genetics
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood-2008-12-192054

Hematopoietic stem cells (HSCs) are generally defined by their dual properties of pluripotency and extensive self-renewal capacity. However, a lack of experimental clarity as to what constitutes extensive self-renewal capacity coupled with an absence of methods to prospectively isolate long-term repopulating cells with defined self-renewal activities has made it difficult to identify the essential components of the self-renewal machinery and investigate their regulation. We now show that cells capable of repopulating irradiated congenic hosts for 4 months and producing clones of cells that can be serially transplanted are selectively and highly enriched in the CD150+ subset of the EPCR+CD48−CD45+ fraction of mouse fetal liver and adult bone marrow cells. In contrast, cells that repopulate primary hosts for the same period but show more limited self-renewal activity are enriched in the CD150− subset. Comparative transcriptome analyses of these 2 subsets with each other and with HSCs whose self-renewal activity has been rapidly extinguished in vitro revealed 3 new genes (VWF, Rhob, Pld3) whose elevated expression is a consistent and selective feature of the long-term repopulating cells with durable self-renewal capacity. These findings establish the identity of a phenotypically and molecularly distinct class of pluripotent hematopoietic cells with lifelong self-renewal capacity.