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A limited temporal window for the derivation of multilineage repopulating
hematopoietic progenitors during embryonal stem cell differentiation in
vitro
N Hole, GJ Graham, U Menzel and JD Ansell
Institute of Cell Animal and Population Biology, University of Edinburgh,
Scotland.
Embryonal stem cells have been shown to differentiate in vitro into all
hematopoietic lineages. This has been used successfully as one approach to
the study of genetic events occurring during haematopoiesis. However,
studies on the commitment of mesodermal precursors to the hematopoietic
lineage have been limited due to the inability to define a system in which
embryonal stem (ES) cells will give rise to primitive hematopoietic stem
cells in vitro. Using a colony forming assay (CFU- A), we determined that
the earliest time point at which primitive multilineage hematopoietic
precursors can be detected during ES cell differentiation in vitro in the
absence of exogenous conditioned medium or stromal cell culture is 4 days.
Lethally irradiated adult recipient mice that received differentiated ES
cells from this time point survived for more than 3 weeks; and in two out
three experiments, peripheral blood from these animals contained ES-derived
progeny. Fluorescence activated cell sorting (FACS) found ES-derived CD45+
hematopoietic cells in both lymphoid and myeloid compartments at 12 weeks
posttransplantation, suggesting that the population of day 4 differentiated
ES cells contains primitive hematopoietic precursors. A preliminary RT-PCR
analysis of gene expression around this time point suggests that there are
very few hematopoietic cells present. This approach should prove useful in
studies of genetic control of commitment to and maintenance of
hematopoietic lineages in vitro and in vivo.
Volume 88,
Issue 4,
pp. 1266-1276,
08/15/1996
Copyright © 1996 by The American Society of Hematology
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