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Blood, Vol. 93 No. 4 (February 15), 1999:
pp. 1253-1263
Maturation of Embryonic Stem Cells Into Endothelial Cells in an
In Vitro Model of Vasculogenesis
Masanori Hirashima,
Hiroshi Kataoka,
Satomi Nishikawa,
Norihisa Matsuyoshi, and
Shin-Ichi Nishikawa
From the Department of Molecular Genetics, Department of Geriatric
Medicine, and Department of Dermatology, Graduate School of Medicine,
Kyoto University, Kyoto, Japan.
A primitive vascular plexus is formed through coordinated regulation
of differentiation, proliferation, migration, and cell-cell adhesion of
endothelial cell (EC) progenitors. In this study, a culture system was
devised to investigate the behavior of purified EC progenitors in
vitro. Because Flk-1+ cells derived from ES cells did not
initially express other EC markers, they were sorted and used as EC
progenitors. Their in vitro differentiation into ECs, via vascular
endothelial-cadherin (VE-cadherin)+ platelet-endothelial
cell adhesion molecule-1 (PECAM-1)+ CD34
to VE-cadherin+ PECAM-1+
CD34+ stage, occurred without exogenous factors, whereas
their proliferation, particularly at low cell density, required OP9
feeder cells. On OP9 feeder layer, EC progenitors gave rise to
sheet-like clusters of Flk-1+ cells, with VE-cadherin
concentrated at the cell-cell junction. The growth was suppressed by
Flt-1-IgG1 chimeric protein and dependent on vascular endothelial
growth factor (VEGF) but not placenta growth factor (PIGF). Further
addition of VEGF resulted in cell dispersion, indicating the role of
VEGF in the migration of ECs as well as their proliferation. Cell-cell
adhesion of ECs in this culture system was mediated by VE-cadherin.
Thus, the culture system described here is useful in dissecting the
cellular events of EC progenitors that occur during vasculogenesis and
in investigating the molecular mechanisms underlying these processes.
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