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CD34+ endothelial cell lines derived from murine yolk sac induce the
proliferation and differentiation of yolk sac CD34+ hematopoietic
progenitors
C Fennie, J Cheng, D Dowbenko, P Young and LA Lasky
Department of Immunology, Genentech, Inc, South San Francisco, CA 94080,
USA.
Embryonic hematopoiesis is initiated in part in the blood islands of the
yolk sac. Previous confocal microscopic analysis has shown that the CD34
antigen, a mucin-like cell surface glycoprotein that is expressed by
hematopoietic progenitors and all endothelial cells of the adult and
embryo, is also found on a subset of luminal hematopoietic-like cells in
the yolk sac blood islands as well as on the vascular endothelium lining
these early hematopoietic locations. We show here that, as in all other
hematopoietic sites thus far examined, immunoaffinity- purified CD34+
nonadherent cells from murine yolk sacs contain the vast majority of
erythroid and myeloid progenitor cell colony forming activity. To examine
the developmental interactions between these CD34+ hematopoietic progenitor
cells of the yolk sac and the CD34+ yolk sac endothelium, we have
immunaffinity-purified adherent endothelial cells from day 10.5 yolk sacs
using CD34 antiserum and produced cell lines by transformation with a
retrovirus expressing the polyoma middle T antigen. Analysis of these cell
lines for CD34, von Willebrand's factor, FLK 1 and FLT 1 expression, and
capillary growth in Matrigel indicates that they appear to be endothelial
cells, consistent with their original phenotype in vivo. Coculture of yolk
sac CD34+ hematopoietic cells on these endothelial cell lines results in up
to a 60-fold increase in total hematopoietic cell number after
approximately 8 days. Analysis of these expanded hematopoietic cells showed
that the majority were of the monocyte/macrophage lineage. In addition,
examination of the cultures showed the rapid formation of numerous
cobblestone areas, a previously described morphologic entity thought to be
representative of early pluripotential stem cells. Scrutiny of the ability
of these endothelial cell lines to expand committed progenitor cells showed
up to a sixfold increase in erythroid and myeloid colony- forming cells
after 3 to 6 days in culture, consistent with the notion that these
embryonic endothelial cells mediate the expansion of these precursor cells.
Polymerase chain reaction analyses showed that most of the cell lines
produce FLK-2/FLT-3 ligand, stem cell factor, macrophage colony-stimulating
factor, leukemia-inhibitory factor, and interleukin- 6 (IL-6), whereas
there is a generally low or not measurable production of granulocyte
colony-stimulating factor, granulocyte-macrophage colony- stimulating
factor, IL-1, IL-3, transforming growth factor beta-1, erythropoietin, or
thrombopoietin. The output of mature hematopoietic cells from these
cocultures can be modified to include an erythroid population by the
addition of exogenous erythropoietin. These data suggest that endothelial
cell lines derived form the yolk sac provide an appropriate hematopoietic
environment for the expansion and differentiation of yolk sac progenitor
cells into at least the myeloid and erythroid lineages.
Volume 86,
Issue 12,
pp. 4454-4467,
12/15/1995
Copyright © 1995 by The American Society of Hematology
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