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Adhesion of NFS-60 myeloid leukemia cells to MC3T3-G2/PA6 stromal cells
induces granulocyte colony-stimulating factor production
T Yoshikubo, K Ozawa, K Takahashi, M Nishikawa, N Horiuchi, A Tojo, K Tani, H Kodama and S Asano
Department of Hematology-Oncology, University of Tokyo, Japan.
To examine the interaction between immature myeloid cells and stromal cells
in the induction of granulocyte colony-stimulating factor (G-CSF)
production, stromal cells of the MC3T3-G2/PA6 (PA6) murine cell line, which
has preadipocyte characteristics and can support hematopoiesis, were
cocultured with various myeloid cell lines and G-CSF mRNA expression was
examined by Northern and reverse transcriptase- polymerase chain reaction
analyses. A significant amount of G-CSF mRNA was induced by the culture of
an interleukin-3/G-CSF-dependent murine myeloid leukemia cell line, NFS-60,
on PA6 stromal cells for 16 hours. Using a G-CSF-dependent subline of DA-1
(DA-1N), the biologic activity of G-CSF was also detected in PA6/NFS-60
coculture supernatants, but not in the culture supernatant of PA6 or NFS-60
alone. Direct contact of NFS-60 cells with the PA6 stromal layer was
essential for the induction of G-CSF mRNA, as indicated by the following
observations: (1) NFS-60 cells efficiently adhered to PA6 cells; (2) medium
conditioned by NFS-60 cells did not contain the activity to induce G- CSF
mRNA in PA6 cells; and (3) induction of G-CSF mRNA was not observed when
NFS-60 cells were separated from PA6 cells by a microporous membrane
(0.45-microns pore size). Several other myeloid cell lines, including
FDC-P2, 32Dcl3, WEHI-3, and DA-1, did not induce G-CSF mRNA expression
after the coculture with PA6 cells, although significant numbers of these
cells adhered to PA6 cells. Therefore, NFS-60 cells may express or
overexpress a molecule that is involved in adhesion- mediated induction of
G-CSF production.
Volume 84,
Issue 2,
pp. 415-420,
07/15/1994
Copyright © 1994 by The American Society of Hematology
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