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A Bigas, DI Martin and ID Bernstein
Pediatric Oncology Program, Fred Hutchinson Cancer Research Center 98109,
USA.
Murine embryonic stem cells are able to differentiate into embryoid bodies
(EBs) in vitro in the absence of leukemia-inhibitory factor with the
formation of different types of hematopoietic precursors within these EBs.
With the aim of determining the in vitro requirements for the continued
development of hematopoietic colony-forming cells (CFCs) and their progeny
from embryonic stem-derived cells, cells from EBs disrupted after 9 days of
formation in the absence of leukemia- inhibitor factor were cultured under
different conditions. Low numbers of day-9 EB cells (5 x 10(5) or less)
cultured in the presence of several growth factors (interleukin-3 [IL-3],
IL-1, c-kit ligand, basic fibroblast growth factor, insulin growth
factor-1, IL-6, granulocyte colony-stimulating factor, fetal liver kinase-2
ligand) develop few or no CFCs after 1 week of culture. When these cells
are plated on irradiated NIH-3T3 with IL-3 or c-kit ligand or combinations
containing these and other growth factors, they are able to generate CFCs
for at least 3 weeks. These cultures were found to include granulocytic,
monocytic, erythrocytic, and megakaryocytic cells. Transwell cultures in
which NIH-3T3 cells were separated from the EB cells and cultures in which
cells were replaced by NIH-3T3 conditioned medium showed that the
interaction between EB-derived cells and NIH-3T3 is via a soluble
factor(s). These studies show that maximal generation of hematopoietic CFCs
from precursors present in day-9 EBs is stimulated by a combination of
known hematopoietic growth factors and a soluble factor(s) produced by
NIH-3T3 cells.
This article has been cited by other articles:
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| Copyright © 1995 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
