Aerosols as a Source of Widespread Mycoplasma Contamination of Tissue Cultures1

O'Connell, Robert C.; Wittler, Ruth G.; Faber, John E.

doi:10.1128/aem.12.4.337-342.1964

Cited by 24 publications

(8 citation statements)

References 7 publications

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“…The contaminant, once established in a cell culture, could easily be transmitted to other cultures by aerosolization. O'Connell et al (124,125) demonstrated that aerosols were created in tissue bottles during trypsinization. When a burette was used to add medium to freshly trypsinized mycoplasmainfected cultures and then was used to add medium to mycoplasma-free cultures, 50 to 75% of them became contaminated.…”

Section: Sources Of Mycoplasma Contami-mentioning

confidence: 99%

Mycoplasmas and cell cultures.

Stanbridge¹

1971

Bacteriological Reviews

118

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Such terms are now in disfavor and the correct generic name, Mycoplasma, should be used. Recently the mycoplasmas have been classified into two separate families, the sterol-requiring Mycoplasmataceae and the sterol-nonrequiring Acholeplasmataceae (44). To the present time, over 30 named species of mycoplasmas have been included within the two families. In addition there are several unidentifiable isolates (L. Hayflick, personal communication). The above classification refers to classic (large-colony) mycoplasmas. It does not include T-strain mycoplasmas (so called, because they form tiny colonies approximately one-fourth the size of classic mycoplasma colonies) on agar. T-strain mycoplasmas were first isolated by Shepard from human urino-genital tracts (170). They have now been isolated from various animals. The T-strain group, as a whole, is unclassified at the present time. A unique requirement for urea (171), in addition to sterol requirement, for growth of these mycoplasmas has prompted the suggestion that they should be included as a separate genus in the order Mycoplasmatales (156a). MORPHOLOGY OF MYCOPLASMAS The mycoplasma organism is distinctive for its small size of 130 to 300 nm and for the lack of a cell wall, the limiting boundary being a plasma 206

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Section: Sources Of Mycoplasma Contami-mentioning

confidence: 99%

Mycoplasmas and cell cultures.

Stanbridge¹

1971

Bacteriological Reviews

118

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“…Weekly PPLO tests detect contaminated bottles before the infection can spread to others. Hayflick (5) and O'Connell et al (10) suggest that most contaminants are of human origin, and are spread by faulty handling techniques which result in aerosol sprays spreading the mycoplasma. Great effort was made to improve our methods of cell culture by following recognized procedures.…”

Section: Discussionmentioning

confidence: 99%

“…One of the problems is in recognizing the mycoplasma infection. As O'Connell et al (10) pointed out, as many as 105 to 107 organisms per milliliter may be present without showing noticeable turbidity of the medium or gross cytopathology of the cells. Macpherson (8) maintains that, although it is widely recognized that mammalian cell line cultures may be contaminated with mycoplasmas, few workers know if their cultures are contaminated, nor take the steps to find out.…”

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confidence: 99%

Control of Pleuropneumonia-like Organisms in Cell Culture

Buskirk¹

1967

Applied Microbiology

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Mammalian cell culture systems were maintained free of mycoplasmas by using a 3-day agar plate test as a weekly routine to monitor the conditions of the cells. If contaminated cell cultures were found, they were discarded and replaced from a pleuropneumonia-like organism (PPLO)-free cell bank. PPLO-free lines were established by treatment with various antibiotics. The KB cell line was freed of mycoplasmas by treatment for 1 week with a mixture of chlortetracycine, kanamycin, and chloramphenicol. L-929 cells were cleared of contamination with either spectinomycin or tylosin, and a synovial cell line was cleared with lincomycin or tylosin. Each cell line, after eradication of the contaminant, was stored in liquid nitrogen. A number of agents were tested to determine minimal inhibitory concentration against three known and three unidentified mycoplasmas. Chlortetracycline and tetracycline were found to be highly active against all strains, whereas tylosin, spectinomycin, and lincomycin, though less active, were equally useful because of their low toxicity against cells. Kanamycin was highly active against three strains, but inactive at high levels against the KB cell contaminants. A disc plate test was used to check isolated cell contaminants for sensitivity to various agents.

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“…M. orale 941 produces CPE in at least two different tissue-culture systems. Tissue-culture contamination by mycoplasma may originate from the oral cavity (O'Connell, Wittler, and Faber, 1964;Hayflick, 1965b), and M. orale is a common oral isolate (Taylor-Robinson et al., 1964). Thus, the finding of an Al.…”

Section: Discussionmentioning

confidence: 99%

Suppression of Rous Sarcoma Virus Growth in Tissue Cultures byMycoplasma orale

Somerson

Cook

1965

J Bacteriol

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Suppression of Rous sarcoma virus growth in tissue cultures by Mycoplasma orale. J. Bacteriol. 90:534-540. 1965.-An agent which produced cell destruction in human diploid and chick-embryo fibroblasts was isolated from WI-26 strain of human diploid fibroblasts and shown to be a mycoplasma. The multiplication of Rous sarcoma virus (RSV) and Rous associated virus (RAV) was inhibited in WI-26, WI-38, and chick-embryo fibroblasts infected with this mycoplasma. The mycoplasma isolate, designated strain 941, reacted strongly in the complement-fixation test with antiserum to Mycoplasma orale CH19299, an isolate obtained from the human oral cavity. The cytopathic effect of mycoplasma strain 941 could be eliminated by growing the mycoplasma on an artificial agar medium before inoculation into chick-embryo fibroblasts. Serial passage in chick-embryo fibroblasts restored the cytopathogenicity of the agar-grown mycoplasma. However, growth of RSV and R1AV was inhibited by both the tissue culture-grown and the agar-grown 941 strain, and also by the CH19299 strain which did not produce any cytopathic effect. IRSV Schmidt-Ruppin (S-R) strain antibodies. Complement-fixation tests. All antigen detection tests were done by use of the micro CF method (Sever, 1962; Huebner et al., 1963) with 4 uniits of anitibody and 2 units of complement. Production of hamster RSV, SIt antiserum, and antigens,

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Aerosols as a Source of Widespread Mycoplasma Contamination of Tissue Cultures1

Cited by 24 publications

References 7 publications

Mycoplasmas and cell cultures.

Mycoplasmas and cell cultures.

Control of Pleuropneumonia-like Organisms in Cell Culture

Suppression of Rous Sarcoma Virus Growth in Tissue Cultures byMycoplasma orale

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