The Survival and Function in Vitro of Nondividing Alveolar Macrophages under Standard Culture Conditions

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Abstract
Alveolar macrophages were collected from the lungs of F344 and HMT rats, CBA mice, and baboons by bronchoalveolar lavage. The cells were cultured as a nondividing population in 96-well plates under standardized conditions. The F344 macrophages cultured in medium containing Approsera I survived better than those cultured with autologous serum. However, this difference did not affect the rate of intracellular dissolution of 57Co3O4 particles. Macrophages from all the species studied were well maintained when cultured with Approsera 1, with 60% or more surviving for between 10 days (HMT rats) and 15 days (CBA mice). Throughout this time nearly all the surviving cells remained viable, as judged by propidium iodide exclusion, lysosomal uptake of acridine orange, phagocytosis of 1.75-μm latex beads, and ultrastructural appearance. Thus uniform culture conditions have been established for alveolar macrophages that will permit interspecies comparisons of their function in vitro and of the cytotoxicity of ingested particles.