A Method for Direct Demonstration of Pleuropneumonia-Like Organisms in Cultured Cells.

Fogh, Jørgen; Fogh, H

doi:10.3181/00379727-117-29731

Cited by 109 publications

(39 citation statements)

References 1 publication

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“…day 4 to 5 after infection. To conclude, the morphological changes in our experiments were in all essentials similar to those observed by Fogh & Fogh (1964 with different mycoplasma-cell systems. DISCUSSION It has been known for a long time that mycoplasmas may be closely attached to the living cells so that they cannot be separated from them.…”

Section: Morphological Aspectssupporting

confidence: 64%

“…Cultures of infected cells on coverslips were washed three times in phosphate buffered saline, pH 7 3, to remove extracellular mycoplasmas, and then the cells were stained with orcein by a similar method to that of Fogh & Fogh (1964). The coverslips were examined under the microscope at magnifications from x 400 to x 3200.…”

Section: Microscopical Demonstration Of Mycoplasma Effects On Cell Cumentioning

confidence: 99%

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Quantitative aspects ofMycoplasma pneumoniae-cell relationships in cultures of lung diploid fibroblasts

Larin

Saxby

Buggey

1969

J. Hyg.

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SUMMARYThe FH strain ofMycoplasma pneumoniaewas studied in experimentally infected cultures of lung diploid fibroblasts. The multiplicity of infection was found to have a profound influence on the distribution of the cell-associated and extra-cellular mycoplasmas. At a high multiplicity there was very little difference between the ratio of cell-associated and extracellular mycoplasmas. In contrast, with a low multiplicity the cell-associated mycoplasmas predominated and with further reductions of the inoculum only the cell-associated mycoplasmas could be detected. The significance of these findings is discussed. It is suggested that caution has to be exercised in the determination of the potency of antibiotics against mycoplasma by methods employing acellular media alone, and it is desirable to utilize also appropriate cell culture models. A possible approach to designing such models is provided by this study.We express our appreciation to the Tissue Culture Laboratory, Pfizer Limited, Sandwich, for kindly supplying us with cell cultures. We furthermore thank the Board of Directors, Pfizer Limited, for permission to publish this study. The helpful discussions with our colleagues Drs D. Jackson and G. M. Williamson in preparing the manuscript are acknowledged with thanks.

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Section: Morphological Aspectssupporting

confidence: 64%

Section: Microscopical Demonstration Of Mycoplasma Effects On Cell Cumentioning

confidence: 99%

Quantitative aspects ofMycoplasma pneumoniae-cell relationships in cultures of lung diploid fibroblasts

Larin

Saxby

Buggey

1969

J. Hyg.

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“…(Table 3) and stained preparations did not reveal any abnormalities. The myeoplasma could not be detected by staining with intensified Ciemsa (Coodburn and Marmion, 1962) or Orcein (Fogh and Fogh, 1964). Chromosome patterns of contaminated cells did not differ significantly from those found in uninfected cells.…”

Section: Antibiotics or Chemicals Used In Treating Cell Eulturesmentioning

confidence: 86%

Detection and Treatment of Contaminating Mycoplasmas in Cell Culture

Cross

Goodman

Shaw

1967

Aust J Exp Biol Med

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Summary.Aspects of the problem of myeoplasma contamination of eell eultures have been examined. Particular attention has been paid to the effective eradication of myeoplasma contaminants. The effects of tylosin, a new antibiotic, are examined in detail.The work also describes attempts to eradicate a particularly resistant strain of M. orale type I, which has been isolated from contaminated cell lines in this laboratory. INTRODUGTION.In recent years use has been made of cell cultures as a method for the isolation of mycoplasmas from clinical specimens originating from patients with leukaemia, rheumatoid arthritis or Reiter's disease (Murphy and Furtado, 1963; Negroni, 1964;Grace, Horoszewicz, Stim and Mirand, 1963; Grace, Horoszewicz, Stim, Mirand and James, 1965;Bartholomew and Himes, 1964;Bartholomew, 1965). The validity of this technique relies heavily on the methods of detecting mycoplasmas in cell culture so as to ensure that they are free of contaminating mycoplasmas at the time of use.The isolation or detection of myeoplasma contaminants of cell cultures is not an easy matter. Many methods have been proposed for the detection of mycoplasmas in cell cultures. These have included the direct visualization of myeoplasma by staining of contaminated cell sheets (Fogh and Fogb, 1964; Glyde, 1961), the use of chemical methods based on the presence of arginine dehydrolase in mycoplasmas (Barile and Schimke, 1963;House and Waddell, 1966) and the culture of supernatant fluid or disrupted cells in suitably supplemented agar medium. Methods of detection of myeoplasma in cell cultures have been reviewed in detail by Hayflick (1965).

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“…Experimental cells were subcultured in 35-or 60-mm plates with medium containing 10% fetal bovine serum and no antibiotics. The cells were mycoplasma free as determined by staining (8). The medium was changed every 48 hr.…”

Section: Methodsmentioning

confidence: 99%

Na and K dependence of the Na/K pump in cystic fibrosis fibroblasts.

Reznik¹,

Schneider²,

Mendoza³

1981

Proc. Natl. Acad. Sci. U.S.A.

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The Na and K dependence of the Na/K pump was measured in skin fibroblasts from patients with cystic fibrosis and age/sex-matched controls. Under basal conditions, there was no difference between control and cystic fibrosis cells in protein per cell, intracellular Na and K content, or Na/K pump activity (measured as ouabain-sensitive 86Rb uptake). There was no difference in the Na dependence of the Na/K pump between cystic fibrosis cells and control cells. In cells from patients with cystic fibrosis, the Na/K pump had a significantly lower affinity for K (Km = 1.6 mM) when compared to normals (Km = 0.9 mM). This difference was demonstrated by using two independent experimental designs.

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A Method for Direct Demonstration of Pleuropneumonia-Like Organisms in Cultured Cells.

Cited by 109 publications

References 1 publication

Quantitative aspects ofMycoplasma pneumoniae-cell relationships in cultures of lung diploid fibroblasts

Quantitative aspects ofMycoplasma pneumoniae-cell relationships in cultures of lung diploid fibroblasts

Detection and Treatment of Contaminating Mycoplasmas in Cell Culture

Na and K dependence of the Na/K pump in cystic fibrosis fibroblasts.

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