title: Differential gene expression profiling of adult murine hematopoietic stem cells.
abstract: 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.
authors.affiliation: University of Michigan, Department of Internal Medicine, Ann Arbor, USA.
authors.forename: In-Kyung
authors.initials: IK
authors.lastname: Park
authors.forename: Yaqin
authors.initials: Y
authors.lastname: He
authors.forename: Fangming
authors.initials: F
authors.lastname: Lin
authors.forename: Ole D
authors.initials: OD
authors.lastname: Laerum
authors.forename: Qiang
authors.initials: Q
authors.lastname: Tian
authors.forename: Roger
authors.initials: R
authors.lastname: Bumgarner
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Klug
authors.forename: Kaijun
authors.initials: K
authors.lastname: Li
authors.forename: Christian
authors.initials: C
authors.lastname: Kuhr
authors.forename: Michelle J
authors.initials: MJ
authors.lastname: Doyle
authors.forename: Tao
authors.initials: T
authors.lastname: Xie
authors.forename: Michèl
authors.initials: M
authors.lastname: Schummer
authors.forename: Yu
authors.initials: Y
authors.lastname: Sun
authors.forename: Adam
authors.initials: A
authors.lastname: Goldsmith
authors.forename: Michael F
authors.initials: MF
authors.lastname: Clarke
authors.forename: Irving L
authors.initials: IL
authors.lastname: Weissman
authors.forename: Leroy
authors.initials: L
authors.lastname: Hood
authors.forename: Linheng
authors.initials: L
authors.lastname: Li
mesh_terms: 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
chemical_list: D018076:DNA, Complementary; D011506:Proteins; D012333:RNA, Messenger