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Murine spleen stromal cell line SPY3-2 maintains long-term hematopoiesis in
vitro
J Tsuchiyama, M Mori and S Okada
Department of Pathology, Okayama University Medical School, Japan.
The hematopoietic microenvironment (HIM) of mouse spleen predominantly
induces the differentiation of hematopoietic progenitors into erythroid
lineage in vivo. However, the mechanisms of this phenomenon have not been
fully explored because of the lack of an adequate in vitro system mimicking
the spleen hematopoiesis. To reconstruct the HIM of mouse spleen in vitro,
we established spleen stromal cell lines from a three- dimensional (3D)
spleen primary culture in collagen gel matrix. Of these, SPY3-2 cells were
negative for preadipocytic and endothelial markers, had a fibroblastoid
morphology, and were not converted to adipocytes in the presence of 1
mumol/L hydrocortisone. They supported the maintenance and multilineal
differentiation of hematopoietic progenitor cells for more than 8 weeks in
vitro. The differentiated hematopoietic cells in the coculture medium were
predominantly monocytes rather than granulocytes. Furthermore,
erythropoiesis was predominantly induced in the presence of 2 U/mL
erythropoietin and continued for more than 12 weeks. The number of
burst-forming units- erythroid (BFU-E) was increased 10 times after 3 weeks
of coculture, which was followed by pronounced production of erythroid
cells in the coculture after week 4. SPY3-2 expressed high levels of c-kit
ligand and low levels of granulocyte macrophage colony-stimulating factor
and interleukin-3, and these molecules were all involved in this long-term
erythropoiesis. Thus, the clonal SPY3-2 cell line will provide a novel HIM
in vitro analogous to that of mouse spleen in vivo. These results suggest
that 3D collagen gel culture may facilitate the establishment of
functioning stromal cell lines of hematopoietic organ.
Volume 85,
Issue 11,
pp. 3107-3116,
06/01/1995
Copyright © 1995 by The American Society of Hematology
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