PURPOSE: To obtain the subject macrophage having a high cell-injuring activity against a wide range of tumor cells by isolating monocytes from a human peripheral blood and subsequently culturing the monocytes in the presence of interleukin-2 and a specific cytokinin in a human serum-containing nutritive medium.
CONSTITUTION: A human peripheral blood is centrifuged to remove erythrocytes, and the obtained buffy coat is treated with a percol solution to isolate highly pure monocytes. The monocytes are cultured in the presence of interleukin-2 and a cytokinin (e.g. colony-stimulating factor-1) relating to the differentiation or activation of monocyte/macrophage in a human serum-containing nutritive medium at 37°C in 5% carbon dioxide atmosphere for 6-17 days, and the cytokinin-activated macrophage produced in the cultured solution is washed with a phosphoric acid buffer solution and subsequently mixed with a 2mM ethylenediamine tetraacetic acid buffer solution. The floated cells are recovered to give the objective cytokinin-activated macrophages.
AZUMA NOBUAKI
OSAWA TOSHIAKI
