1887

Abstract

A simple, fast, and method of detecting the inapparent infection of cultured cells with mycoplasmas is reported. Animal cells grown on Formvar-coated electron microscopic grids were directly fixed with glutaraldehyde, negatively stained with phosphotungstic acid and examined by transmission electron microscopy. Cells contaminated with mycoplasmas could be discriminated from uncontaminated cells. The micro-organisms in the negatively stained preparations corresponded with those revealed by thin sectioning, and the distribution of mycoplasmas in cultured cells coincided with those revealed by the Hoechst staining method. Most of the highly resolved mycoplasmas were polymorphic, and closely associated with host cells; often more than 500 organisms per host cell were seen.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-129-11-3379
1983-11-01
2021-05-09
Loading full text...

Full text loading...

/deliver/fulltext/micro/129/11/mic-129-11-3379.html?itemId=/content/journal/micro/10.1099/00221287-129-11-3379&mimeType=html&fmt=ahah

References

  1. Barile M.F., Schrimke R.T. 1963; A rapid chemical method for detecting PPLO contamination of tissue cell cultures. Proceedings of the Society for Experimental Biology 114:676–679
    [Google Scholar]
  2. Barile M.F., Malizia W., Riggs D.B. 1962; Incidence and detection of pleuropneumonia-like organisms in cell cultures by fluorescent antibody and cultural procedures. Journal of Bacteriology 84:130–136
    [Google Scholar]
  3. Boatman E.S., Kenny G.E. 1971; Morphology and ultrastructure of Mycoplasma pneumoniae spherules. Journal of Bacteriology 106:1005–1015
    [Google Scholar]
  4. Brown S., Teplitz M., Revel J.-P. 1974; Interaction of mycoplasmas with cell cultures, as visualized by electron microscopy. Proceedings of the National Academy of Sciences of the United States of America 71:464–468
    [Google Scholar]
  5. Buckley I.K., Porter K.R. 1975; Electron microscopy of critical point dried whole cultured cells. Journal of Microscopy 104:107–120
    [Google Scholar]
  6. Chen T.R. 1977; In situ detection of mycoplasma contamination in cell cultures by fluorescent Hoechst 33258 stain. Experimental Cell Research 104:255–262
    [Google Scholar]
  7. Hakala M.T., Holland J.F., Horoszewicz J.S. 1963; Change in pyrimidine deoxyribonucleoside metabolism in cell culture caused by mycoplasma (PPLO) contamination. Biochemical and Biophysical Research Communications 11:466–471
    [Google Scholar]
  8. Ichimaru H., Nakamura M. 1979; The components of Mycoplasma salivarium and its growth medium that are responsible for film formation. Journal of General Microbiology 112:389–392
    [Google Scholar]
  9. Katsumoto T., Takayama H., Takagi A. 1978; Ultrastructural organization of cultured macrophages as shown by negative staining technique. Journal of Electron Microscopy 27:1–11
    [Google Scholar]
  10. Kimura G., Itagaki A. 1975; Initiation and maintenance of cell transformation by simian virus 40: a viral genetic property. Proceedings of the National Academy of Sciences of the United States of America 72:673–677
    [Google Scholar]
  11. Macpherson I. 1966; Mycoplasmas in tissue culture. Journal of Cell Science 1:145–168
    [Google Scholar]
  12. Nardone R.M., Todd J., Gonzalez P., Gaffney E.V. 1965; Nucleoside incorporation into strain L cells: inhibition by pleuropneumonia-like organisms. Science 149:1100–1101
    [Google Scholar]
  13. Schneider E.L., Stanbridge E.J., Epstein C.J., Golbus M., Abbo-Halbasch G., Rodgers G. 1974; Mycoplasma contamination of cultured amniotic fluid cells: potential hazard to prenatal chromosomal diagnosis. Science 184:477–480
    [Google Scholar]
  14. Stanbridge E. 1971; Mycoplasmas and cell cultures. Bacteriological Reviews 35:206–227
    [Google Scholar]
  15. Zucker-Franklin D., Davidson M., Thomas L. 1966; The interaction of mycoplasmas with mammalian cells. I. HeLa cells, neutrophils, and eosinophils. Journal of Experimental Medicine 124:521–541
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-129-11-3379
Loading
/content/journal/micro/10.1099/00221287-129-11-3379
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error