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Blood, Vol. 95 No. 7 (April 1), 2000:
pp. 2275-2283
Vascular endothelial growth factor synergistically enhances bone
morphogenetic protein-4-dependent lymphohematopoietic cell generation
from embryonic stem cells in vitro
Naoki Nakayama,
Jae Lee, and
Laura Chiu
From the Department of Cell Biology and the Flow Cytometry
Laboratory, Amgen Inc, Thousand Oaks, CA.
The totipotent mouse embryonic stem (ES) cell is known to
differentiate into cells expressing the  -globin gene when stimulated with bone morphogenetic protein (BMP)-4. Here, we demonstrate that
BMP-4 is essential for generating both erythro-myeloid colony-forming cells (CFCs) and lymphoid (B and NK) progenitor cells from ES cells and
that vascular endothelial growth factor (VEGF) synergizes with BMP-4.
The CD45+ myelomonocytic progenitors and
Ter119+ erythroid cells began to be detected with 0.5 ng/mL BMP-4, and their levels plateaued at approximately 2 ng/mL.
VEGF alone weakly elevated the CD34+ cell
population though no lymphohematopoietic progenitors were induced.
However, when combined with BMP-4, 2 to 20 ng/mL VEGF synergistically
augmented the BMP-4-dependent generation of erythro-myeloid CFCs and
lymphoid progenitors from ES cells, which were enriched in
CD34+ CD31lo and CD34+
CD45 cell populations, respectively, in a dose-dependent
manner. Furthermore, during the 7 days of in vitro differentiation,
BMP-4 was required within the first 4 days, whereas VEGF was functional
after the action of BMP-4 (in the last 3 days). Thus, VEGF is a
synergistic enhancer for the BMP-4-dependent differentiation processes,
and it seems to be achieved by the ordered action of the 2 factors.
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