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Initiation of hematopoiesis and vasculogenesis in murine yolk sac explants
J Palis, KE McGrath and PD Kingsley
Department of Pediatrics, University of Rochester Medical Center, NY 14642,
USA.
The blood islands of the visceral yolk sac (VYS) are the initial sites of
hematopoiesis in mammals. We have developed a yolk sac explant culture
system to study the process of blood cell and endothelial cell development
from extraembryonic mesoderm cells. No benzidine-positive cells or beta
H1-globin mRNA expression was detected at the primitive streak or neural
plate stage of development (E7.5). However, when isolated E7.5 dissected
tissues were cultured for 36 to 72 hours in serum-free medium, hundreds of
hemoglobin-producing cells and embryonic globin gene expression were
identified in both intact yolk sac and VYS mesoderm explants. Explanted
E7.5 extraembryonic mesoderm tissues thus recapitulate in vivo primitive
erythropoiesis and do not require the presence of a vascular network or the
VYS endoderm. Yolk sac blood islands also contain endothelial cells that
arise by vasculogenesis and express flk-1. We detected flk-1 mRNA as early
as the primitive streak stage of mouse embryogenesis. Culture of embryo
proper and intact VYS explants, which contain both mesoderm and endoderm
cells, produced capillary networks and expressed flk-1. In contrast,
vascular networks were not seen when VYS mesoderm was cultured alone,
although flk-1 expression was similar to that of intact VYS explants. The
addition of vascular endothelial growth factor to VYS mesoderm explants did
not induce vascular network formation. These results suggest that the VYS
endoderm or its extracellular matrix is necessary for the coalescence of
developing endothelial cells into capillary networks.
Volume 86,
Issue 1,
pp. 156-163,
07/01/1995
Copyright © 1995 by The American Society of Hematology
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