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Blood, Vol. 92 No. 11 (December 1), 1998:
pp. 4108-4118
GATA-1 Regulates Growth and Differentiation of Definitive Erythroid
Lineage Cells During In Vitro ES Cell Differentiation
Naruyoshi Suwabe,
Satoru Takahashi,
Toru Nakano, and
Masayuki Yamamoto
From the Center for Tsukuba Advanced Research Alliance and Institute
of Basic Medical Sciences, University of Tsukuba, Tsukuba, Japan; and
Institute of Microbial Diseases, Osaka University, Osaka, Japan.
Although the importance of GATA-1 in both primitive and definitive
hematopoietic lineages has been shown in vivo, the precise roles played
by GATA-1 during definitive hematopoiesis have not yet been clarified.
In vitro differentiation of embryonic stem (ES) cells using OP9 stroma
cells can generate primitive and definitive hematopoietic cells
separately, and we have introduced a method that separates
hematopoietic progenitors and differentiated cells produced in this
system. Closer examination showed that the expression of erythroid
transcription factors in this system is regulated in a differentiation
stage-specific manner. Therefore, we examined differentiation of GATA-1
promoter-disrupted (GATA-1.05) ES cells using this system.
Because the GATA-1.05 mice die by 12.5 embryonic days due to
the lack of primitive hematopoiesis, the in vitro analysis is an
important approach to elucidate the roles of GATA-1 in definitive
hematopoiesis. Consistent with the in vivo observation, differentiation
of GATA-1.05 mutant ES cells along both primitive and
definitive lineages was arrested in this ES cell culture system. Although the maturation-arrested primitive lineage cells did not express detectable amounts of  y-globin mRNA, the
blastlike cells accumulated in the definitive stage showed  -globin
mRNA expression at approximately 70% of the wild type. Importantly,
the TER119 antigen was expressed and porphyrin was accumulated in the
definitive cells, although the levels of both were reduced to
approximately 10%, indicating that maturation of definitive erythroid
cells is arrested by the lack of GATA-1 with different timing from that of the primitive erythroid cells. We also found that the hematopoietic progenitor fraction of GATA-1.05 cells contains more
colony-forming activity, termed CFU-OP9. These results suggest that the
GATA-1.05 mutation resulted in proliferation of
proerythroblasts in the definitive lineage.
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