Hematopoietic Fingerprints: an expression database of stem cells and their progeny. Mus musculus

Hematopoietic stem cells (HSC) continuously regenerate a complete hematologic and immune system. Very few genes that regulate this process have yet been identified. In order to identify factors governing differentiation, we have compared the transcriptome of highly purified HSC with their differentiated progeny, including erythrocytes, granulocytes, monocytes, NK cells, activated and naïve T-cells, and B-cells. Chromosomal analysis revealed that HSC were more transcriptionally active than other cell types across most chromosomes. Each lineage expressed ~100 to 400 genes uniquely, including many previously uncharacterized genes. Overexpression of two fingerprint genes resulted in a significant bias in differentiation indicating a role in cell fate determination, demonstrating the utility of these data for modulation of specific cell types. Keywords: Hematopoietic cells, stem cells, wild type cells, Mus musculus Overall design: Goal - Expression profiling of hematopoietic cells Description - Biological replicates of 10 hematopoietic cell types from female C57Bl/6 mice were purified for microarray analysis Keywords: Hematopoietic cells, stem cells, wild type cells, Mus musculus Experimental Factors: wild-type female C57Bl/6 CD45.1 Mus musculus Experimental Design: 25,000- 50,000 cells were FACS sorted, mRNA was purified by RNAqueous (Ambion), DNAse treated with DNAse1, phenol:chloroform:isoamyl alcohol extracted, and amplified through two rounds of in vitro transcription using MessageAmp (Ambion). Second round was biotinylated with bio-CTP and bio-UTP. Microarrays: Oligonucleotide, Affymetrix, Moe430 v 2.0 QC: Biological replicates, correlation coefficient > .95; scale factor <10; 5'/3' ratio < 20