Long-Term Cultivation of Mouse Peritoneal Macrophages12

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Abstract
Four procedures were considered essential for the long-term cultivation of peritoneal macrophages: 1) The peritoneal exudate was washed out with the chemically defined, protein-free medium NCTC 109 without further centrifugation or washing to preserve the nutrients for growth and to prevent the loss of vital substances through dialysis; 2) 40 percent horse serum and 50 percent NCTC 109 served as the nutrient medium; 3) the medium was supplemented with 10 percent beef-embryo extract diluted 1:5; and 4) cultures were maintained in an atmosphere of 5 percent CO2 in air which regulated the medium at a pH of about 7.2. Mitotic division of macrophages was observed in the early stages of cultures and up to 43 days, but probably occurred infrequently in the older cultures. Macrophages were maintained in good condition for 220 days. The results on the various steps of development of the present cell system are presented.