THE IN VITRO DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES

Abstract

Mouse mononuclear phagocytes cultivated in 50% newborn calf serum medium pinocytize actively and form large numbers of phase-dense granules as well as 3 hydrolytic enzymes. When such cells are then placed in 1% newborn calf serum they illustrate a low level of pinocytic activity, a shrinkage in granule size, and a loss in cell protein, acid phosphatase, [beta]-glucuronidase, and cathepsin. Examination of the extracellular medium revealed no detectable hydrolase activity. The reintroduction of cells into high levels of serum again resulted in granule and enzyme formation. Cells rapidly incorporated fluorescein-conjugated calf serum proteins into the phase-dense granules. The fluorescence of labeled granules was lost during an 18 hour period in non-fluorescein-containing medium. Crystalline egg white lysozyme was concentrated in the macrophages. Approximately 80% of the cell-associated enzyme was lost during a 24 hour washout period in either 1 or 50% serum medium. No enzymatic activity could be recovered in the medium. Colloidal gold was taken up and concentrated in macrophage granules. Quantitative assays revealed this particle to be conserved during a 24 hour washout period.