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Fibroblasts induce heparin synthesis in chondroitin sulfate E containing
human bone marrow-derived mast cells
L Gilead, O Bibi and E Razin
Institute of Biochemistry, Hebrew University-Hadassah Medical School,
Jerusalem, Israel.
Human bone marrow-derived mast cells (hBMMCs), differentiated in vitro in
suspension culture and under the influence of human peripheral blood
mononuclear cells conditioned medium (hCM), were tested for their response
to recombinant human interleukin-3 (rhIL-3) and for their behavior in
different microenvironments. The hBMMCs were incubated in the presence of
rhIL-3 and the changes in their proliferation rate were determined.
Recombinant hIL-3 induced a more than sixfold increase in 3H-thymidine
uptake into the hBMMC DNA in a dose-dependent manner. Human CM used as a
control for proliferation response induced a more than eightfold maximal
proliferation rate increase. Rabbit anti-rhIL-3 completely inhibited hBMMC
3H-thymidine uptake induced by rhIL-3 and decreased the hCM-induced
proliferation by approximately 50%. These hBMMCs were cocultured with four
different mytomicin C-treated cell monolayers and assayed for phenotypic
changes. After only 2 days in coculture with either embryonic mouse
skin-derived fibroblasts (MESFs) or human skin-derived fibroblasts (HSFs),
a marked increase in granule number and density was noted on staining with
toluidine blue. Mast cells that initially stained alcian blue+/safranin- at
day 0 of coculture became alcian blue+/safranin+ during the coculture
period. Human BMMC proteoglycan synthesis shifted from approximately 85%
chondroitin sulfate E to approximately 60% heparin within 14 to 19 days of
coculture with the MESF monolayer and to approximately 50% heparin within
19 days of coculture with the HSF monolayer. None of the above- mentioned
changes were noted in cocultures of hBMMCs with 3T3 cell line fibroblast
monolayers or in cocultures with bovine vascular endothelium (BVE) cell
monolayers. These results demonstrate microenvironmental effects exerted by
the MESF and HSF monolayers on IL-3-dependent hBMMCs similar to those
reported in the conversion of murine mast cell phenotype.
Volume 76,
Issue 6,
pp. 1188-1195,
09/15/1990
Copyright © 1990 by The American Society of Hematology
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