Journal of Leukocyte Biology 44:111-121 (1988) © 1988 Alan R. Liss, Inc. Long-Term Cultivation of Functional Human Macrophages in Teflon Dishes With Serum-Free Media

Reported herein are the results of studies demonstrating the utility of a chemically defined, serum-free medium designated as AIM-V (G1BCO) for the long-term (>2 weeks) cultivation of functIonally-defined human macrophages. The AIM-V medium isa mixture of HEPES-buffered Dulbecco’s Modified Eagle Medium and Ham’s Nutrient Mixture F12 that had been supplemented with purified human albumin, transferrin, Insulin, and a proprietary mixture of purIfied factors. Nonadherent macrophages for serial studies were generated In petri dishes with a Teflon lIner that had been seeded with a heteroge-neous population of peripheral biood mononuclear cells of healthy human adults. For comparison, cultures were initiated with serum-free medium AIM-V and medium supple-mented with AB/Rh + serum or freshly collected autologous serum. Viability was defined by trypan blue dye, glass adherence, and phagocytosis of yeast. Macrophages were characterized by light microscopy, cytochemistry, and phenotypic analysis. Ultrastruc-tural morphology was defined by scanning electron microscopy. These studies demon-strate that functionally defined human macrophages can be sustained in long-term culture with the use of serum-free medium that has not been augmented with mitogenic stimulants, growth-promoting lymphokineslmonekines, or differentiation-inducing agents. Serum-free medium AIM-V which has been approved for generating lymphokine, (I.e., lnterleukln-2; lL-2)-actlvated killer cells (LAK) for lL-2ILAK adoptive immunotherapy modalities, may also prove useful for studies defining and Isolating regulatory proteins produced by activated monocytes and macrophages and for generating cytolytic mac-rophages for different antitumor regImens. Key words: monocyte, cultureware, phagocytosls, scanning electron microscop