Gene expression profile of primary human CD34+CD38lo cells differentiating along the megakaryocyte lineage.
OBJECTIVE
To identify genes involved in megakaryopoiesis, high-density oligonucleotide microarrays were used to compare transcript profiles from undifferentiated CD34+CD38lo cells and culture-derived megakaryocytes (MKs).


METHODS
Megakaryocyte differentiation was achieved in vitro by inducing primary human CD34+CD38lo cells in serum-deprived media supplemented with the cytokine combination of interleukin-3, interleukin-6, stem cell factor, and thrombopoietin for 10 days. Three replicate microarray experiments were performed using hematopoietic cells isolated from three different organ donors and high-density oligonucleotide microarrays.


RESULTS
Analysis of gene array data resulted in 304 differentially expressed genes (p < or = 0.001, fold change > or = 3). A third of the 25 most highly up-regulated genes were known to participate in hemostasis (z = 6.75), and no genes known to be associated with MKs were among the down-regulated genes. We also found a large proportion of up-regulated transcripts in gene ontology categories of adhesion and receptor activity (85%) and signal transduction activity (68%). At the same time, 70% of genes within transcription factor functions were down-regulated. Confirmatory studies indicated that the array results correlated with mRNA and protein expression levels in primary MKs.


CONCLUSIONS
This study provides a global expression profile of human MKs and a list of novel and previously uncharacterized candidate genes that are important components of megakaryopoiesis.
Shim
Mi-Hyun
MH
Puget Sound Blood Center, Seattle, WA 98104, USA. mihyuns@psbc.org
Hoover
Aubree
A
Blake
Noel
N
Drachman
Jonathan G
JG
Reems
Jo Anna
JA
D036541:ADP-ribosyl Cyclase / Q000097:blood*; D051997:ADP-ribosyl Cyclase 1; D015703:Antigens, CD / Q000097:blood*; D018952:Antigens, CD34 / Q000097:blood*; D001483:Base Sequence; D002454:Cell Differentiation; D004247:DNA / Q000097:blood / Q000235:genetics; D017931:DNA Primers; D005434:Flow Cytometry / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000276:immunology*; D006801:Humans; D016130:Immunophenotyping; D008533:Megakaryocytes / Q000166:cytology / Q000276:immunology*; D008562:Membrane Glycoproteins; D020411:Oligonucleotide Array Sequence Analysis; D011003:Ploidies
D015703:Antigens, CD; D018952:Antigens, CD34; D017931:DNA Primers; D008562:Membrane Glycoproteins; D004247:DNA; D036541:ADP-ribosyl Cyclase; C498154:CD38 protein, human; D051997:ADP-ribosyl Cyclase 1