1. What are Mycoplasmas?
Mycoplasmas are very small bacteria lacking cell walls that belong to various genera within the class Mollicutes. They are flexible, pleomorphic organisms that can be as small as 0.2 to 0.3.
2. What are the consequences of mycoplasma contamination?
Because of their typically high concentrations in cell cultures, mycoplasmas can often out compete with the host cells for essential nutrients which result in altered growth and production of proteins. After all, the number of mycoplasma in a contaminated culture can easily outnumber the cells by a factor of 1000:1 or more. Usually there is an absence of visible morphological changes but the invisible effects on the cell cultures are what make mycoplasma contamination so serious a problem.
3. I have used the Mycoplasma Elimination Cocktail on my cells but my cells are dying. Why is this happening?
The mycoplasma cocktail is a very potent combination of different antibiotics to increase the success of eliminating mycoplasma. Even though we have tested and confirmed no cytotoxicity was observed for a variety of cell lines, we would still recommend optimizing the best dilution for your specific cell line in order to achieve best results.
4. Which mycoplasma strains are targeted?
Potential strains which may be affected are: Mycoplasma pneumonia strains Mycoplasma hyosynoviae strains Mycoplasma hyorhinis strains Mycoplasma synoviae strains Note: Some strains may naturally become tolerant to this cocktail due to their endogenous defense mechanisms, and thus may show reduced sensitivity. In such cases, other antibiotic combinations may need to be considered
5. Is it possible to do a second treatment with Saveit ?
Yes, it is possible to run a second treatment after the first passage. The first treatment reduces the mycoplasma load drastically. The second treatment would kill remaining mycoplasma.
6. What is the principle of Saveit ? How does it kill mycoplasma and how long will mycoplasma survive in general?
Saveit contains membrane active components acting in combination with the alcohols included. It also includes aldehydes mainly for the inactivation of non-enveloped viruses and spores. Saveit is also active against bacteria and other microorganisms. It is not quite understood how long mycoplasma can survive outside a culture or their natural habitat. By our knowledge no study is available. It is only known that cross contamination is very prominent for cell cultures. That can only be the case with quite stable mycoplasma.
7. Does Saveit interfere with ELISA applications?
No, Saveit does not interfere with any type of ELISA parameter due to the low molecular weight of the active component. It's the mycoplasma and their effects on the cells providing, after their removal, real results with the ELISA.
8. What are recommended methods of eliminating mycoplasma contamination on surfaces or other laboratory apparatus?
Saveit Surface Disinfection Spray is ideal for cleansing and disinfection of all laboratory surfaces and apparatus including clean benches, incubators, work benches, cell storage boxes and liquid nitrogen containers. Saveit is also effective against a broad range of other pathogenic contaminants.
9. Can primary cells be treated with Saveit ?
Yes, primary cells can be treated with Saveit. However, we recommend a 10-fold increase in the cell concentration. (Note that an overall decrease in the elimination efficiency of Saveit should subsequently be expected).
10. Is Saveit suitable for removing mycoplasma from murine ES cells?
Murine ES cells can be treated with Saveit. Saveit generally shows a low cytotoxicity in contrast to other anti-mycoplasma reagents, but still is not completely free of such complications. We usually like to recommend Saveit which shows basically no cytotoxicity anymore, but treatment takes longer.
11. Can Saveit eliminate intracellular contaminants?
Saveit does not integrate into the cellular membrane. Therefore it cannot eliminate intracellular contamination. However, mycoplasma is an extracellular contaminate. Mycoplasma penetrans and Mycoplasma gallisepticum are the only species described intracellulary. Both M. penetrans and M. gallisepticum have not been reported as cell contaminants.
12. When can I be certain that Mycoplasma is permanently eliminated?
In case that a few mycoplasma particles survive the treatment with Saveit they will grow to detectable titers after four passages. You can detect mycoplasma at an early stage with the highly sensitive Mycoplasma Detection Kit to exclude persisting contamination.