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Fig. 1. Effect of proinflammatory cytokines on the behaviour of 2A4 mammary epithelial cells grown in collagen gels. The cells were suspended in a gelling collagen solution. After a 10 minute incubation at 37°C to allow collagen gelation, defined medium with or without the indicated cytokines was added above the gels and renewed after 3 days. (A) 2A4 cells grown in a collagen gel for 6 days under control conditions form compact ball-like colonies. (B) 2A4 cells treated for 6 days with IL-1ß (2 ng ml-1) form loose cell clusters. (C) 2A4 cells treated for 6 days with TNF-
(10 ng ml-1) do not form cohesive colonies but scatter throughout the collagen matrix, mostly as individual cells. Scale bars, 50 µm. (D) Quantification of the effects of IL-6, IL-1, IL-1ß and TNF- was carried out after 6 days of treatment by counting the number of apparently single cells in five randomly selected fields per experimental condition in each of three independent experiments. Data were expressed as mean number of cells ± s.e.m. and statistical significance was determined using the Student's unpaired t test. P=0.025 for values of IL-6 compared with controls; P<0.0005 for values of IL-1, IL-1ß and TNF- compared with controls, as well as for values of TNF- compared with IL-6, IL-1, IL-1ß.
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