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Frequently Asked Questions, Mycoplasma Testing
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 Why should I think about Mycoplasma testing?
Contamination of cell cultures by Mycoplasma species has been a known risk of cell culture since the 1950s. In a 1964 study, Mycoplasma-contaminated flasks were handled and samples were collected from the surrounding environment to determine their mode of transmission (O’Connell et al. 1964, Appl Microbiol 12: 337-342). It was found that Mycoplasma organisms from contaminated cultures spread primarily through liquid droplets and aerosols induced within the culture flask. A single organism was sufficient to cause contamination within a second culture. Good cell culture practice should reduce the incidence of Mycoplasma contamination, but nevertheless, contamination occurs and spreads all too easily.
How frequent is Mycoplasma contamination? Published figures vary widely, but the average frequency would seem to be 1% of primary cultures, 5% of early passage cultures and 15-35% of established cell lines (Drexler and Uphoff 2002, Cytotechnology 39: 75-90).
It is possible to eliminate a Mycoplasma contaminant using a variety of commercially available agents (antibiotics such as penicillin and streptomycin only serve to mask contamination and cannot be used in treatment). However, repositories continue to advise against continued passaging of contaminated cells unless the culture itself is irreplaceable. Because many Mycoplasma species are located intracellularly, this provides a reservoir for infection, and so the contaminant is hard to eliminate completely. Prevention, with routine testing of all cell lines, is the best cure. |
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