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Blood, Vol. 95 No. 3 (February 1), 2000:
pp. 870-878
Characterization of hematopoietic lineage-specific gene
expression by ES cell in vitro differentiation induction system
Takumi Era,
Toshiaki Takagi,
Tomomi Takahashi,
Jean-Christophe Bories, and
Toru Nakano
From the Department of Molecular Cell Biology, Research Institute
for Microbial Diseases, Osaka University, Osaka, Japan; and Unité
462 Institut National de la Santé et de la Recherche
Médicale, Hôpital Saint-Louis, Paris, France.
The continuous generation of mature blood cells from hematopoietic
progenitor cells requires a highly complex series of molecular events.
To examine lineage-specific gene expression during the differentiation
process, we developed a novel method combining LacZ reporter gene analysis with in vitro
hematopoietic differentiation induction from mouse embryonic stem
cells. For a model system using this method, we chose the erythroid and
megakaryocytic differentiation pathways. Although erythroid and
megakaryocytic cells possess distinct functional and morphologic
features, these 2 lineages originate from bipotential
erythro-megakaryocytic progenitors and share common lineage-restricted
transcription factors. A portion of the 5' flanking region of the
human glycoprotein IIb ( IIb) integrin gene extending from base
 598 to base +33 was examined in detail. As reported previously,
this region is sufficient for megakaryocyte-specific gene expression.
However, previous reports that used human erythro-megakaryocytic cell
lines suggested that one or more negative regulatory regions were
necessary for megakaryocyte-specific gene expression. Our data clearly
showed that an approximately 200-base enhancer region extending from
598 to 400 was sufficient for megakaryocyte-specific gene
expression. This experimental system has advantages over those using
erythro-megakaryocytic cell lines because it recapitulates normal
hematopoietic cell development and differentiation. Furthermore, this
system is more efficient than transgenic analysis and can easily
examine gene expression with null mutations of specific genes.
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