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[A micromethod for measuring macrophage chemotactic factor(s) in the supernatants of mouse lymphocyte cultures stimulated by concanavalin A].

A micromethod is described for the measurement of the macrophage chemotactic factor (s) released by activated murine lymphocytes in culture. Lymphocytes are stimulated by various doses of concanavalin A in the wells of a tissure culture microplate. Two wells are used for thymidine incorporation measurement and three wells for the chemotactic assay. For this assay, microchambers are prepared from segments of a disposable tuberculine syringe closed on oneend by gluing a small Millipore filter. These chambers are placed in the wells of the tissue culture plate so that the filter is just in contact with the supernatant of the lymphocyte culture. They are then filled with 5.104 macrophages obtained from the peritoneum of normal mice suspended in 100 mul of medium 199 supplemented with bovine serum albumin. The macrophages migrate into the filter more deeply in experiments with stimulate lymphocytes than in experiments with non-stimulated cultures. The distance travelled by the macrophage is measured by the difference between the microscope focal adjustments for the top of the filter and the deepest migrating cells respectively.