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Blood, Vol. 93 No. 5 (March 1), 1999:
pp. 1549-1556
In Vitro Hematopoietic and Endothelial Cell Development From Cells
Expressing TEK Receptor in Murine Aorta-Gonad-Mesonephros Region
Isao Hamaguchi,
Xu-Ling Huang,
Nobuyuki Takakura,
Jun-ichi Tada,
Yuji Yamaguchi,
Hiroaki Kodama, and
Toshio Suda
From the Department of Cell Differentiation, Institute of Molecular
Embryology and Genetics, Kumamoto University School of Medicine,
Kumamoto; and Research Center Kyoto, Bayer Yakuhin Ltd, Kyoto, Japan.
Recent studies have shown that long-term repopulating hematopoietic
stem cells (HSCs) first appear in the aorta-gonad-mesonephros (AGM)
region. Our immunohistochemistry study showed that TEK+
cells existed in the AGM region. Approximately 5% of AGM cells were
TEK+, and most of these were CD34+ and
c-Kit+. We then established a coculture system of AGM
cells using a stromal cell line, OP9, which is deficient in macrophage
colony-stimulating factor (M-CSF). With this system, we showed that AGM
cells at 10.5 days postcoitum (dpc) differentiated and proliferated
into both hematopoietic and endothelial cells. Proliferating
hematopoietic cells contained a significant number of colony-forming
cells in culture (CFU-C) and in spleen (CFU-S). Among primary AGM cells at 10.5 dpc, sorted TEK+ AGM cells generated
hematopoietic cells and platelet endothelial cell adhesion molecule
(PECAM)-1+ endothelial cells on the OP9 stromal layer,
while TEK cells did not. When a ligand for TEK,
angiopoietin-1, was added to the single-cell culture of AGM,
endothelial cell growth was detected in the wells where hematopoietic
colonies grew. Although the incidence was still low (1/135), we showed
that single TEK+ cells generated hematopoietic cells and
endothelial cells simultaneously, using a single-cell deposition
system. This in vitro coculture system shows that the
TEK+ fraction of primary AGM cells is a candidate for
hemangioblasts, which can differentiate into both hematopoietic cells
and endothelial cells.
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