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. 1965 Sep 1;122(3):455–466. doi: 10.1084/jem.122.3.455

THE IN VITRO DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES

III. THE REVERSIBILITY OF GRANULE AND HYDROLYTIC ENZYME FORMATION AND THE TURNOVER OF GRANULE CONSTITUENTS

Zanvil A Cohn 1, Belinda Benson 1
PMCID: PMC2138068  PMID: 5320303

Abstract

Mouse mononuclear phagocytes cultivated in 50 per cent newborn calf serum medium pinocytize actively and form large numbers of phase-dense granules as well as three hydrolytic enzymes. When such cells are then placed in 1 per cent newborn calf serum they illustrate (a) a low level of pinocytic activity, (b) a shrinkage in granule size, and (c) a loss in cell protein, acid phosphatase, β-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 per cent of the cell-associated enzyme was lost during a 24 hour washout period in either 1 or 50 per cent 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.

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Selected References

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  1. COHN Z. A., BENSON B. THE DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. MORPHOLOGY, CYTOCHEMISTRY, AND BIOCHEMISTRY. J Exp Med. 1965 Jan 1;121:153–170. doi: 10.1084/jem.121.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. COHN Z. A., BENSON B. THE IN VITRO DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. I. THE INFLUENCE OF INHIBITORS AND THE RESULTS OF AUTORADIOGRAPHY. J Exp Med. 1965 Feb 1;121:279–288. doi: 10.1084/jem.121.2.279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. COHN Z. A., BENSON B. THE IN VITRO DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. II. THE INFLUENCE OF SERUM ON GRANULE FORMATION, HYDROLASE PRODUCTION, AND PINOCYTOSIS. J Exp Med. 1965 May 1;121:835–848. doi: 10.1084/jem.121.5.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. COHN Z. A., HIRSCH J. G. The isolation and properties of the specific cytoplasmic granules of rabbit polymorphonuclear leucocytes. J Exp Med. 1960 Dec 1;112:983–1004. doi: 10.1084/jem.112.6.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. COHN Z. A., WIENER E. THE PARTICULATE HYDROLASES OF MACROPHAGES. I. COMPARATIVE ENZYMOLOGY, ISOLATION, AND PROPERTIES. J Exp Med. 1963 Dec 1;118:991–1008. doi: 10.1084/jem.118.6.991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. GOSSELIN R. E. The uptake of radiocolloids by macrophages in vitro; a kinetic analysis with radioactive colloidal gold. J Gen Physiol. 1956 May 20;39(5):625–649. doi: 10.1085/jgp.39.5.625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. HARFORD C. G., HAMLIN A., PARKER E. Electron microscopy of HeLa cells after the ingestion of colloidal gold. J Biophys Biochem Cytol. 1957 Sep 25;3(5):749–756. doi: 10.1083/jcb.3.5.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. HOLTZER H., HOLTZER S. The in vitro uptake of fluorescein labelled plasma proteins. I. Mature cells. C R Trav Lab Carlsberg. 1960;31:373–408. [PubMed] [Google Scholar]
  9. RIGGS J. L., SEIWALD R. J., BURCKHALTER J. H., DOWNS C. M., METCALF T. G. Isothiocyanate compounds as fluorescent labeling agents for immune serum. Am J Pathol. 1958 Nov-Dec;34(6):1081–1097. [PMC free article] [PubMed] [Google Scholar]

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