A large number of mouse fibrosarcoma and adult guinea pig fibroblast cultures were examined for their ability to produce migration inhibitory activity. In most cases culture supernatants were found to inhibit macrophage migration, in a dose-dependent manner. Toxicity of the tested material could be excluded by: a) experiments using colchicine as a stimulator of macrophage migration and, b) examination of the effect of test materials on macrophage monolayer cultures. Additionally, migration inhibitory activity was found in fibroblast, but not fibrosarcoma frozen and thawed extracts. Furthermore, incubation of cells with puromycin could only inhibit production by fibrosarcoma, thus suggesting that the fibroblast activity was due to preformed cellular constituents. Fractionation of concentrated culture supernatants by Sephadex G-200 gel filtration showed that the activity derived from fibrosarcoma cells could be eluted in a narrow molecular weight fraction (18,000-22,500), whereas the fibroblast activity was heterogenously distributed over a wide range. Migration inhibitory activity in fibroblast extracts was mainly associated with higher molecular weight material. Differences could be demonstrated between these activities and lymphocyte migration inhibitory factor, including inhibition by methyl-pentoses and the presence of macrophage aggregating activity.
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