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Erythropoiesis and vasculogenesis in embryoid bodies lacking visceral yolk
sac endoderm
M Bielinska, N Narita, M Heikinheimo, SB Porter and DB Wilson
Department of Pediatrics, Washington University School of Medicine, St
Louis, MO, USA.
During mouse embryogenesis the first hematopoietic and endothelial cells
form in blood islands located between layers of visceral endoderm and
mesoderm in the yolk sac. The role of visceral endoderm in primitive
hematopoiesis and vasculogenesis is not well understood. We have assessed
the consequences of a lack of visceral endoderm on blood cell and vessel
formation using embryoid bodies derived from mouse embryonic stem (ES)
cells deficient in GATA-4, a transcription factor expressed in yolk sac
endoderm. When differentiated in vitro, these mutant embryoid bodies do not
develop an external visceral endoderm layer. We found that Gata4-/-
embryoid bodies, grown either in suspension culture or attached to a
substratum, are defective in primitive hematopoiesis and vasculogenesis as
evidenced by a lack of recognizable blood islands and vascular channels and
a reduction in the expression of the primitive erythrocyte marker epsilon
y-globin. Expression of the endothelial cell transcripts FIk-1, FIt-1, and
platelet-endothelial cell adhesion molecule (PECAM) was not affected in the
mutant embryoid bodies. Gata4-/- ES cells retained the capacity to
differentiate into primitive erythroblasts and endothelial cells when
cultured in methylcellulose or matrigel. Analysis of chimeric mice,
generated by injecting Gata4-/- ES cells into 8-cell stage embryos of
ROSA26 transgenic animals, showed that Gata4-/- ES cells can form blood
islands and vessels when juxtaposed to visceral endoderm in vivo. We
conclude that the visceral endoderm is not essential for the
differentiation of primitive erythrocytes or endothelial cells, but this
cell layer plays an important role in the formation and organization of
yolk sac blood islands and vessels.
Volume 88,
Issue 10,
pp. 3720-3730,
11/15/1996
Copyright © 1996 by The American Society of Hematology
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