Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of mouse primitive and maturing hematopoietic stem cells


ABSTRACT: Gene expression studies from hematopoietic stem cell (HSC) populations purified to variable degrees have defined a set of stemness genes. The present study describes the construction and comparative molecular analysis of l-phage cDNA libraries from highly purified primitive HSCs (PHSCs) which retained their long term repopulating activities (LTRAs), and from maturing HSCs (MHSCs) which were largely depleted of LTRAs. Library inserts were amplified and tagged by a T7 RNA polymerase promoter and used to generate biotinylated cRNA for Microarray hybridization. Microarray analysis of the libraries confirmed previous results but also revealed an unforseen preferential expression of translation and metabolism associated genes in the PHSCs. Therefore these data indicate that HSCs are quiescent only in regard of proliferative activities, but are in a state of readiness to provide the metabolic and translational activities required following induction of proliferation by factors which induce differentiation and exit from the HSC pool. We used microarrays to detail the global programme of gene expression distinguishing primitive and maturing hematopoietic stem cells from mouse bone marrow and identified distinct classes of up- and down-regulated genes. Experiment Overall Design: To compare the transcriptosomes of primitive and maturing hematopoietic stem cells from mouse bone marrow, cDNA libraries were generated from RNA isolated from highly purified stem cell populations and used to generate biotinylated cRNA for Affymetrix microarray analysis.

ORGANISM(S): Mus musculus

SUBMITTER: Ludger Klein-Hitpass 

PROVIDER: E-GEOD-4391 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Gene expression profiles in murine hematopoietic stem cells revisited: analysis of cDNA libraries reveals high levels of translational and metabolic activities.

Hüttmann Andreas A   Dührsen Ulrich U   Heydarian Katja K   Klein-Hitpass Ludger L   Boes Tanja T   Boyd Andrew W AW   Li Chung-Leung CL  

Stem cells (Dayton, Ohio) 20060330 7


Gene expression studies from hematopoietic stem cell (HSC) populations purified to variable degrees have defined a set of stemness genes. Unexpectedly, results also hinted toward a HSC chromatin poised in a wide-open state. With the aim of providing a robust tool for further studies into the molecular biology of HSCs, the studies herein describe the construction and comparative molecular analysis of lambda-phage cDNA libraries from highly purified HSCs that retained their long-term repopulating  ...[more]

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