Preparation of effector cells

1 Resuspend monocytes/macrophages at 1 x 106 cells/ml in complete RPMI 1640 medium.

2 Plate 0.1 ml of the macrophage cell suspension (containing 1 x 105 cells) into triplicate wells of a 96-well, flat-bottom microtitre tissue culture plate.

3 Add the appropriate macrophage activators in 0.1 ml of medium to yield the final volume of 0.2 ml/well (see Protocol 1). Incubate the cells for 18-24 h at 37°C with activators. Centrifuge the plate at 250-300 g for 5 min. Remove supernatant by vacuum aspiration.

Wash the macrophage effector cells in the plate three times with warm complete medium (see Protocol 1).

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