Fibroblasts maintain the phenotype and viability of the rat heparin-containing mast cell in vitro

F. Levi-Schaffer, K. F. Austen, J. P. Caulfield, A. Hein, W. F. Bloes, R. L. Stevens

Research output: Contribution to journalArticlepeer-review

125 Scopus citations

Abstract

Rat serosal heparin-containing mast cells (HP-MC) were maintained in vitro for as long as 30 days when co-cultured with mouse skin-derived 3T3 fibroblasts. In contrast, when the mast cells were cultured alone, on fibronectin-, gelatin-, or dermal-collagen-coated dishes, on acid and heat-killed fibroblasts in the presence or absence of 24 hr fibroblast-conditioned medium, or on a monolayer of mouse serosal macrophages, they failed to adhere to the dishes, released significant amounts of their histamine and lactate dehydrogenase, and stained with trypan blue, indicating a loss of viability. The rat serosal HP-MC cultured with the 3T3 fibroblasts became so adherent to the fibroblasts that the two cell types could be separated from one another only by trypsinization. The cultured HP-MC stained with both alcian blue and safranin and continued to synthesize proteoglycan at a rate comparable to that of freshly isolated cells. The 35S-labeled proteoglycan synthesized by these cultured cells, like that produced by freshly isolated rat serosal HP-MC, was a 750,000 to 1,000,000 m.w. proteoglycan containing only heparin glycosaminoglycans of 50,000 to 100,000 m.w. When HP-MC were cultured for 1 wk with the fibroblasts and were then incubated for 5 min with a 1/20 dilution of rabbit anti-rat IgE, they generated and released an average of 22 ± 10 ng (mean ± SD, n = 5) of prostaglandin D2 per 106 cells and exocytosed a higher net percentage of their total histamine content (44 ± 11% [mean ± SD, n = 8]) than did cells just isolated from the animal (6 ± 4% [mean ± SD, n = 4]). As assessed by electron microscopy, many of the cultured HP-MC resembled freshly isolated cells except that some secretory granules had fused with one another in some cells. Morphologically, after activation the cultured HP-MC underwent compound exocytosis like freshly isolated cells. These results demonstrate that the in vivo differentiated rat HP-MC maintain their histology, morphology, immunologic responsiveness, histamine content, and ability to synthesize heparin proteoglycan when co-cultured with living fibroblasts.

Original languageAmerican English
Pages (from-to)3454-3462
Number of pages9
JournalJournal of Immunology
Volume135
Issue number5
StatePublished - 1985
Externally publishedYes

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