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Prepublished online as a Blood First Edition Paper on July 10, 2003; DOI 10.1182/blood-2003-04-1249. This Article Full Text (PDF) All Versions of this Article: 2003-04-1249v1 102/9/3120    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Klarmann, K. D Articles by Keller, J. R Search for Related Content PubMed PubMed Citation Articles by Klarmann, K. D Articles by Keller, J. R Related Collections Related Article in Blood Online Social Bookmarking                   What's this? Submitted April 22, 2003 Accepted June 23, 2003 Identification of in vitro growth conditions for c-Kit-negative hematopoietic stem cells Kimberly D Klarmann*, Mariaestela Ortiz, Meghan R Davies, and Jonathan R Keller Laboratory of Molecular Immunoregulation, Center for Cancer Research, National Cancer Institute-Frederick, NIH, Frederick, MD, USA Basic Research Program, SAIC-Frederick, Center for Cancer Research, National Cancer Institute-Frederick, NIH, Frederick, MD, USA * Corresponding author; email: kklarmann{at}ncifcrf.gov. Our laboratory recently identified a quiescent class of pluripotent hematopoietic stem cells (PHSC) that are lineage negative (Linneg), lack c-Kit, and are able to give rise to c-Kit positive (c-Kitpos) PHSC in vivo. This population fails to proliferate in vitro, but has delayed reconstituting activity in vivo. In this study, we purified these cells to enrich for the PHSC and we identified in vitro conditions capable of supporting their maturation. The c-Kitneg cells exhibited differential expression of Sca-1, CD34, CD43, CD45, and Thy 1.2. We purified the cells based on Sca-1, as it is expressed on active PHSC. We detected pre-CFU-s activity in both the Sca-1neg and Sca-1pos populations, indicating the presence of primitive PHSC in both populations. However, our in vitro studies suggest the Sca-1pos population is enriched for PHSC. The in vitro systems that support the growth of these dormant cells include a modified long-term marrow culture and various stromal cell lines. In modified long-term bone marrow cultures, c-Kitneg cells gave rise to c-Kitpos PHSC, with long-term reconstitution activity in vivo. Thus we have established an in vitro system to examine PHSC maturation that will allow us to study the mediators of the c-Kitneg to c-Kitpos transition. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Article in Blood Online: What if nothing grows? Gerald J. Spangrude Blood 2003 102: 3077. [Full Text] [PDF] This article has been cited by other articles: F. Cerisoli, L. Cassinelli, G. Lamorte, S. Citterio, F. Bertolotti, M. C. Magli, and S. Ottolenghi Green fluorescent protein transgene driven by Kit regulatory sequences is expressed in hematopoietic stem cells Haematologica, March 1, 2009; 94(3): 318 - 325. [Abstract] [Full Text] [PDF] G. V. Hegde, K. J. Peterson, K. Emanuel, A. K. Mittal, A. D. Joshi, J. D. Dickinson, G. J. Kollessery, R. G. Bociek, P. Bierman, J. M. Vose, et al. Hedgehog-Induced Survival of B-Cell Chronic Lymphocytic Leukemia Cells in a Stromal Cell Microenvironment: A Potential New Therapeutic Target Mol. Cancer Res., December 1, 2008; 6(12): 1928 - 1936. [Abstract] [Full Text] [PDF] B. Jacquelin, T. Kortulewski, P. Vaigot, A. Pawlik, G. Gruel, O. Alibert, P. Soularue, C. Joubert, X. Gidrol, and D. T.-L. Roux Novel pathway for megakaryocyte production after in vivo conditional eradication of integrin {alpha}IIb-expressing cells Blood, September 15, 2005; 106(6): 1965 - 1974. [Abstract] [Full Text] [PDF]

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