Human Blood Monocytes: Characterization of Negatively Selected Human Monocytes and Their Suspension Cell Culture Derivatives

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Abstract
Normal human monocytes were negatively selected from leucapheresis cell suspensions by countercurrent centrifugation–elutriation in high yield with a mean purity of 93.5%. The combination of the novel methods of negative cell selection and suspension cell culture has provided the opportunity to study serially over several days the morphologic and functional changes of monocytes from a single donor as they matured in culture to typical macrophages. Human monocytes nearly double in size during the first week of culture, experiencing near daily increases in cell volume. This was associated with changes in the ultrastructure of these cells, including the development of numerous small knob‐like projections on the cell membrane and the proliferation of microtubules and filamentous Structures within the cell cytoplasm during the first 6 days of culture. Peroxidase activity declined during the first 4 days of culture, whereas 5′‐nucleotidase activity was acquired during the first 48 h of culture. Lysozyme activity in the cultures increased from day 2 to day 6 of culture. The phagocytic capacity of monocytes for IgG‐coated erythrocytes increased dramatically during the first week of culture, but the cytotoxic capability of monocytes against similar targets in an antibody‐dependent cytotoxicity assay declined to nearly hall of base‐line levels by day 2 of culture and remained at this diminished level during subsequent days in culture.

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