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Phagocytosis of mycobacteria by U937 cells: a rapid method for monitoring uptake and separating phagocytosed and free bacteria by magnetic beads
Author(s) -
Whyte J.,
Roberts A. D. G.,
Morley K. A.,
Sharp R. J.,
Marsh P. D.
Publication year - 2000
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.2000.00701.x
Subject(s) - phagocytosis , u937 cell , bacteria , microbiology and biotechnology , mycobacterium phlei , cell sorting , cell culture , biology , flow cytometry , macrophage , chemistry , mycobacterium , biochemistry , in vitro , genetics
J. WHYTE, A.D.G. ROBERTS, K.A. MORLEY, R.J. SHARP and P.D. MARSH.2000.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.