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Purification and characterization of heterogeneous pluripotent
hematopoietic stem cell populations expressing high levels of c-kit
receptor
D Orlic, R Fischer, S Nishikawa, AW Nienhuis and DM Bodine
Clinical Hematology Branch, National Heart, Lung, and Blood Institute,
Bethesda, MD 20892.
Mouse pluripotent hematopoietic stem cells (PHSC) were fractionated based
on size and density using counterflow centrifugal elutriation (CCE). These
heterogeneous PHSC populations were further enriched by subtraction of
cells with lineage-specific markers (Lin-) followed by positive sorting for
c-kit expression. The cells were characterized for their functional and
biochemical properties. We defined a subpopulation of c-kit-positive cells
that expressed high numbers of c-kit receptors (c-kitBR). One hundred
c-kitBR cells from either low- or higher-density fractions were sufficient
to repopulate the lymphohematopoietic system in WBB6F1-W/Wv (W/Wv)
recipients, whereas no PHSC were found in cells with low (c-kitDULL) or no
(c-kitNEG) c-kit expression. Lin- c-kitBR cells were separated into RhoDULL
and RhoBR subsets based on their ability to efflux rhodamine 123 (Rho). The
PHSC were concentrated in Lin- c-kitBR RhoDULL cells and the number of Lin-
c-kitBR RhoBR cells correlated directly with the number of day 12
colony-forming unit- spleen (CFU-S12) in each fraction. We were not able to
enrich further for PHSC using monoclonal antibodies to the cell-surface
markers AA4.1 or CD4, which have been used by others to isolate PHSC. The
small, low- density Lin- c-kitBR subset contained PHSC and few CFU-S12.
This enabled us to assay PHSC for expression of the flk-2 gene, which
encodes a tyrosine kinase receptor present on fetal liver PHSC. Purified
RNA from the low-density Lin- c-kitBR subset did not contain flk-2 mRNA. We
suggest that AA4.1, CD4 and flk-2 are expressed as stage- specific markers
on PHSC in cell cycle.
Volume 82,
Issue 3,
pp. 762-770,
08/01/1993
Copyright © 1993 by The American Society of Hematology
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|
|
|
|
|
|
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|
|
|
|
|
|
|
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|
|
|
|
|
|
|
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|
|
|
|
|
|
|
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January 5, 1997;
124(10):
2039 - 2048.
[Abstract]
[PDF]
|
|
|
|
|
|
|
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January 1, 1997;
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[Abstract]
[Full Text]
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|
|
|
|
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|
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|
|
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