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Mycoplasma Elimination

The contamination of cells with mycoplasmas is a very common problem, even though it often remains unnoticed since no cloudiness appears in the cell culture. Nevertheless, the contamination often causes significant biochemical changes as well as changes in the immunological properties of the cells. For both biological and economical reasons, it is important to eliminate mycoplasmas from cell cultures used for basic research, diagnosis, quality testing, and biotechnological production. Since mycoplasma-infected cells cannot always be discarded, many complicated methods have been suggested for the elimination of the mycoplasmas.

PromoKine is offering two highly effective methods to get rid of mycoplasma contamination:

 

Mycoplasma-EX, a combination of two solutions (non-antibiotic/antibiotic)  with extremely low cytotoxicity. While the antibiotic efficiently inhibiting mycoplasma growth, the non-antibiotic solution directly kills mycoplasmas fast and very efficiently. It can be used to clean most eukaryotic cell cultures or virus stocks directly. Cultures are free of mycoplasmas after only four passages!

BIOMYC, a combination of three different antibiotics which have been shown to be very effective in the elimination of mycoplasma species that account for 90% of the contamination found in cell cultures. When used according to our instructions, no cytotoxic effects will occur. The BIOMYC solutions are a very cost-effective way to get cell cultures free of mycoplasmas - however, it is more time-consuming and labor-intensive compared to the Mycoplasma-EX treatment.

 

Using Antibiotics to Disinfect Cell Cultures

A number of effective methods for the elimination of mycoplasma contamination in cell cultures have been published, such as:

  • treatment with specific hyperimmune serum (antibodies).
  • passage of contaminated cells in thymus-deficient mice.
  • exposure to analogs of nucleic acids that prevent reproduction of mycoplasma.
  • treatment with antibiotics.
  • exposure of contaminated cells to mouse macrophages.
  • a technique that combines growing cells on soft agar and treatment with antibiotics.

The preferred method in terms of simplicity is treatment with antibiotics, which do not damage or alter cells.

Antibiotics such as penicillin, which attacks bacterial cell walls, are ineffective in this instance, since mycoplasmas lacks a true cell wall. Several antibiotics eliminate mycoplasmas effectively, such as: Tylosin, Neomycin, Tetracycline and Gentamycin. However, the efficacy of these antibiotics is restricted to specific mycoplasma species and frequently only reduce the concentration of mycoplasmas rather than disinfect the cell culture. Hence, as soon as treatment is concluded, contamination will recur.

Two methods are recommended for treating contaminated cells with antibiotics. The first is based on alternating treatment with two types of antibiotics, and the second on treatment with one type of antibiotic.