Mycoplasma contamination represents a significant problem to the culture of mammalian cells used for research. Mycoplasmas can cause disastrous effects on eukaryotic cells as they can alter every cellular parameter leading to unreliable experimental results and potentially unsafe biological products. Mycoplasmas cannot be detected by visual inspection using a normal light microscope and thus, can remain unnoticed in the cell cultures for long periods.

Mycoplasma Detection

InvivoGen has developed PlasmoTest™, a cell-based assay that provides a simple, rapid and reliable method to detect all mycoplasma species known to infect cell cultures. More info on Mycoplasma detection

Mycoplasma Elimination

Mycoplasma contamination can be efficiently and rapidly eliminated with the Mycoplasma removal agents, Plasmocin™ orPlasmocure™. They combine two antibiotics that act through different mechanisms and allow mycoplasma eradication in only 2 weeks. 

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Mycoplasma Eradication Review

Mycoplasma contamination in cell cultures

Mycoplasma contamination of cultured cells is a major problem in both basic research and industrial production. Up to 87% of cell lines may be contaminated by mycoplasma. 
Mycoplasma infection can affect virtually any function and activity of eukaryotic cells leading to experimental artifacts and unreliable results.

It is usually recommended that an infected cell culture be immediately autoclaved to prevent the infection from spreading and to use only mycoplasma-free cultures. However, some cell lines are irreplaceable and require an effective eradication treatment.

Antibiotics active against Mycoplasma

The use of specific antibiotics can efficiently eliminate mycoplasma contaminations. Antibiotics commonly used in cell culture are inactive on mycoplasma (e.g. penicillins and streptomycin). 
Three classes of antibiotics have been shown to kill mycoplasma at relatively low concentrations: tetracyclines, macrolides and quinolones. Tetracyclines and macrolides block the protein synthesis by interfering with ribosome translation, while quinolones inhibit the replication of bacterial DNA.

Commercially available anti-mycoplasma agents for cell culture

Several antibiotics are commercially available for the removal of mycoplasma: BM-Cyclin (Roche) contains a macrolide and a tetracycline, Ciprobay (Bayer, available only with a prescription) and MRA (ICN) are both quinolones. Plasmocin™ (InvivoGen) is the only antimycoplasma reagent that combines a macrolide and a quinolone. 
Unlike BM-Cyclin that requires the sequential and cyclic use of two antibiotics, Plasmocin™ is ready-to-use and can be added to the culture medium directly. 
Furthermore, both components in Plasmocin™ act on separate targets blocking protein synthesis and DNA replication, whereas the two antibiotics in BM-Cyclin are both inhibitors of protein synthesis. Therefore, Plasmocin™ is more effective in removing mycoplasma and prevents the appearance of resistant strains. 
In contrast to other anti-mycoplasma compounds, Plasmocin™ is active on both free mycoplasma as well as intracellular forms. This advantage is conferred by one component of Plasmocin™ which is actively transported into mammalian cells. It ensures that following treatment with Plasmocin™ a cell culture is not reinfected by mycoplasma released from intracellular compartments of infected cells. 
To date, no consistent and permanent alterations that affect the eukaryotic cells during and after the treatment have been detected [1].

Comparison of the most common anti-mycoplasma agents

1. Uphoff CC, Drexler HG., 2005. Eradication of mycoplasma contaminations. Methods Mol Biol. 290:25-34.
2. Somasundaram C. et al., 1992. Use of ciprofloxacin and BM-Cyclin in mycoplasma decontamination. In vitro Cell Dev Biol. 28A(11-12):708-10.
3. Drexler HG. et al., 1994. Treatment of mycoplasma contamination in a large panel of cell cultures. In vitro Cell Dev Biol Anim. 30A(5):344-7

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