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Blood, 15 January 2002, Vol. 99, No. 2, pp. 488-498
HEMATOPOIESIS
Differential gene expression profiling of adult murine
hematopoietic stem cells
In-Kyung Park,
Yaqin He,
Fangming Lin,
Ole D. Laerum,
Qiang Tian,
Roger Bumgarner,
Christopher A. Klug,
Kaijun Li,
Christian Kuhr,
Michelle J. Doyle,
Tao Xie,
Michèl Schummer,
Yu Sun,
Adam Goldsmith,
Michael F. Clarke,
Irving L. Weissman,
Leroy Hood, and
Linheng Li
From the University of Michigan, Department of Internal
Medicine, Ann Arbor; the Stowers Institute for Medical Research, Kansas
City, MO; the University of Washington, Departments of Pediatrics,
Microbiology, Surgery, and Molecular Biotechnology, Seattle; the
University of Bergen, Department of Pathology, Norway; the University
of Alabama at Birmingham, Department of Microbiology; Stanford
University, Department of Pathology, Stanford, CA; and the Institute
for Systems Biology, Seattle, WA.
Hematopoietic stem cells (HSCs) have self-renewal capacity and
multilineage developmental potentials. The molecular mechanisms that
control the self-renewal of HSCs are still largely unknown. Here, a
systematic approach using bioinformatics and array hybridization techniques to analyze gene expression profiles in HSCs is described. To
enrich mRNAs predominantly expressed in uncommitted cell lineages, 54 000 cDNA clones generated from a highly enriched population of HSCs
and a mixed population of stem and early multipotent progenitor (MPP)
cells were arrayed on nylon membranes (macroarray or high-density array), and subtracted with cDNA probes derived from mature lineage cells including spleen, thymus, and bone marrow. Five thousand cDNA
clones with very low hybridization signals were selected for sequencing
and further analysis using microarrays on glass slides. Two populations
of cells, HSCs and MPP cells, were compared for differential gene
expression using microarray analysis. HSCs have the ability to
self-renew, while MPP cells have lost the capacity for self-renewal. A
large number of genes that were differentially expressed by enriched
populations of HSCs and MPP cells were identified. These included
transcription factors, signaling molecules, and previously unknown genes.
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