Differential gene expression profiling of adult murine hematopoietic stem cells.
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.
Park
In-Kyung
IK
University of Michigan, Department of Internal Medicine, Ann Arbor, USA.
He
Yaqin
Y
Lin
Fangming
F
Laerum
Ole D
OD
Tian
Qiang
Q
Bumgarner
Roger
R
Klug
Christopher A
CA
Li
Kaijun
K
Kuhr
Christian
C
Doyle
Michelle J
MJ
Xie
Tao
T
Schummer
Michèl
M
Sun
Yu
Y
Goldsmith
Adam
A
Clarke
Michael F
MF
Weissman
Irving L
IL
Hood
Leroy
L
Li
Linheng
L
D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D020869:Gene Expression Profiling; D015723:Gene Library; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000145:classification / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D020411:Oligonucleotide Array Sequence Analysis; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D013382:Subtraction Technique
D018076:DNA, Complementary; D011506:Proteins; D012333:RNA, Messenger