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Changes in hematopoiesis-supporting ability of C3H10T1/2 mouse embryo
fibroblasts during differentiation
M Nishikawa, K Ozawa, A Tojo, T Yoshikubo, A Okano, K Tani, K Ikebuchi, H Nakauchi and S Asano
Department of Hematology-Oncology, University of Tokyo, Japan.
To investigate the functional change of stromal cells along with
differentiation, we used a differentiation-inducible mouse embryo
fibroblast cell line, C3H10T1/2 (10T1/2). Stably determined preadipocyte
and myoblast cell lines were established after a brief exposure of 10T1/2
cells to 5-azacytidine. These cell lines terminally differentiated into
adipocytes and myotubes, respectively, under appropriate conditions. The
hematopoiesis-supporting ability of each 10T1/2-derived cell line was
examined by coculture with FACS-sorted murine hematopoietic stem cells
(Thy-1lo c-kit+ Lin-). The number of granulocyte-macrophage progenitors
(CFU-GM) was slightly reduced after 7 days of culture with parent 10T1/2
fibroblasts, whereas a marked increase in CFU-GM number was observed when
the cells were cultured on preadipocytes. Mature adipocytes and
myogenically determined cell lines, on the other hand, did not support
CFU-GM growth. Further, Northern analysis showed that the preadipocyte cell
line acquired the ability to produce a significant amount of stem cell
factor (SCF), interleukin-6 (IL-6), leukemia inhibitory factor, and
macrophage colony- stimulating factor mRNAs in response to IL-1 or
lipopolysaccharide stimulation. Terminal adipocytic differentiation
resulted in reduced ability to express these cytokine mRNAs. Similarly,
highest IL-6 activity was detected in the supernatant of preadipocyte
culture, whereas adipocytes did not secrete IL-6 even after IL-1
stimulation. Interestingly, hematopoiesis-nonsupporting myoblasts and
myotubes also expressed abundant SCF mRNA, suggesting that SCF, per se, may
not be sufficient for stem cell growth and survival. The 10T1/2-derived
cell lines could provide a valuable tool to aid in the analysis of stromal
cell development and the search for novel stromal factors.
Volume 81,
Issue 5,
pp. 1184-1192,
03/01/1993
Copyright © 1993 by The American Society of Hematology
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