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Murine hematopoietic stem cells change their surface phenotype during ex vivo expansion
Author(s): Zhang CC, Lodish HF
Source: BLOOD    Volume: 105    Issue: 11    Pages: 4314-4320    Published: JUN 1 2005  
Times Cited: 49     References: 37     
Abstract: Ex vivo expansion of hematopoietic stem cells (HSCs) is important for many clinical applications, and knowledge of the surface phenotype of ex vivo-expanded HSCs will be critical to their purification and analysis. Here, we developed a simple culture system for bone marrow (BM) HSCs using low levels of stem cell factor (SCF), thrombopoietin (TPO), insulin-like growth factor 2 (IGF-2), and fibroblast growth factor-1 (FGF-1) in serum-free medium. As measured by competitive re-population analyses, there was a more than 20-fold increase in numbers of long-term (LT)-HSCs after a 10-day culture of total BM cells. Culture of BM "side population" (SP) cells, a highly enriched stem cell population, for 10 days resulted in an approximate 8-fold expansion of repopulating HSCs. Similar to freshly isolated HSCs, repopulating HSCs after culture were positive for the stem cell markers Sca-1, Kit, and CD31 and receptors for IGF-2. Surprisingly, prion protein and Tie-2, which are present on freshly isolated HSCs, were not on cultured HSCs. Two other HSC markers, Endoglin and Mpl, were expressed only on a portion of cultured HSCs. Therefore, the surface phenotype of ex vivo-expanded HSCs is different from that of freshly isolated HSCs, but this plasticity of surface phenotype does not significantly alter their re-population capability.
Document Type: Article
Language: English
Reprint Address: Lodish, HF (reprint author), Whitehead Inst Biomed Res, 9 Cambridge Ctr, Cambridge, MA 02142 USA
Addresses:
1. Whitehead Inst Biomed Res, Cambridge, MA 02142 USA
2. MIT, Dept Biol, Cambridge, MA USA
Publisher: AMER SOC HEMATOLOGY, 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA
Subject Category: Hematology
IDS Number: 928SR
ISSN: 0006-4971
DOI: 10.1182/blood2004-11-4418
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