Coculture of interleukin 3-dependent mouse mast cells with fibroblasts results in a phenotypic change of the mast cells.
The heparin-containing mast cells that reside in the connective tissue of the mouse, but not the chondroitin sulfate-containing mast cells in the gastrointestinal mucosa, stain with safranin when exposed to alcian blue/safranin. Mouse bone marrow-derived mast cells (BMMC), the probable in vitro counterparts of in vivo mucosal mast cells, were cultured for 14 days with mouse skin-derived 3T3 fibroblasts in RPMI 1640 medium containing 10% fetal calf serum and 50% WEHI-3 conditioned medium. Although the BMMC adhered to the fibroblast monolayer, they continued to divide, probably due to the presence of interleukin 3 in the conditioned medium. The mast cells remained viable throughout the period of coculture, since they failed to release lactate dehydrogenase and because they increased their histamine content approximately 15-fold. After 12-14 days of coculture, greater than 50% of the BMMC changed histochemically to become safranin+; 30-40% of the 35S-labeled glycosaminoglycans on the proteoglycans synthesized by these cocultured mast cells were heparin, whereas heparin was not detected in the initial BMMC. In the absence of WEHI-3 conditioned medium, BMMC adhered to the fibroblast monolayer, and after 8 days of coculture, the number of mast cells did not change and their histamine content remained the same. However, these mast cells also became safranin+ and synthesized 40% heparin glycosaminoglycans. Thus, coculture of BMMC with fibroblasts induces a phenotypic change so that the resulting mast cells stain safranin+ and synthesize heparin proteoglycans, whereas the presence of WEHI-3 conditioned medium stimulates proliferation and an increase in histamine content.
Levi-Schaffer
F
F
Austen
K F
KF
Gravallese
P M
PM
Stevens
R L
RL
D000818:Animals; D001854:Bone Marrow Cells; D002448:Cell Adhesion; D002453:Cell Cycle; D002454:Cell Differentiation; D002478:Cells, Cultured; D005347:Fibroblasts / Q000502:physiology*; D006493:Heparin / Q000378:metabolism; D006632:Histamine / Q000378:metabolism; D007377:Interleukin-3; D008222:Lymphokines / Q000502:physiology*; D008407:Mast Cells / Q000166:cytology*; D051379:Mice; D010619:Phenazines; D010641:Phenotype
D007377:Interleukin-3; D008222:Lymphokines; D010619:Phenazines; D006632:Histamine; D006493:Heparin; C009195:safranine T