title: Phagocytosis of mycobacteria by U937 cells: a rapid method for monitoring uptake and separating phagocytosed and free bacteria by magnetic beads.
abstract: A human-derived monocytic cell line (U937) was induced to phagocytose Mycobacterium phlei by the addition of phorbol myristate acetate (PMA) to the culture medium for 50-60 h. Cells not treated with PMA were unable to phagocytose M. phlei. Magnetic beads enabled a rapid and highly efficient separation of phagocytosed and free bacteria to be achieved, an approach which is particularly useful if colony plating is used to enumerate bacterial survival within phagocytic cells. Fluorescence-activated cell sorting (FACS) analysis showed that 98% of U937 cells contained viable bacteria after 3 h.
authors.affiliation: CAMR, Salisbury, Wiltshire, UK.
authors.forename: J
authors.initials: J
authors.lastname: Whyte
authors.forename: A D
authors.initials: AD
authors.lastname: Roberts
authors.forename: K A
authors.initials: KA
authors.lastname: Morley
authors.forename: R J
authors.initials: RJ
authors.lastname: Sharp
authors.forename: P D
authors.initials: PD
authors.lastname: Marsh
mesh_terms: D002469:Cell Separation / Q000379:methods*; D003470:Culture Media; D005434:Flow Cytometry; D006801:Humans; D008280:Magnetics; D008863:Microspheres; D009161:Mycobacterium / Q000276:immunology*; D010587:Phagocytosis; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors; D020298:U937 Cells / Q000187:drug effects / Q000276:immunology* / Q000382:microbiology
chemical_list: D003470:Culture Media; D013755:Tetradecanoylphorbol Acetate