NR ASRZ
AU Zhang,C.C.; Lodish,H.F.
TI Murine hematopoietic stem cells change their surface phenotype during ex vivo expansion
QU Blood 2005 Jun 1; 105(11): 4314-20
PT journal article
AB 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 repopulation 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 repopulation capability.
MH Animals; Antigens, Surface/analysis; Biological Markers/analysis; Bone Marrow Cells/cytology; Cell Culture Techniques; Cell Differentiation; Cell Proliferation; Cells, Cultured; Growth Substances/pharmacology; Hematopoietic Stem Cells/chemistry/*cytology; Immunophenotyping; Mice; Mice, Inbred Strains; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.
AD Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, MA 02142, USA
SP englisch
PO USA