预防和清除支原体污染的细胞培养方法

Since one of the main sources of mycoplasma is the cell cultures brought from outside, it is suggested to supply cells from reliable cell banks.

In the case of the existence of mycoplasma contaminated cell culture in quarantine and the absence of separate incubators, only flasks in a plastic box with lid should be used. Never use plates and unsealed dishes in quarantine. In the case of suspected cultures, handling them at the end of the workday after all other cell culture work is completed, using separated media and reagents, and finally disinfecting the laminar flow hood after working is strongly suggested .

Only use antibiotics responsibly

Although ideally antibiotics used for cell culture should eradicate all contaminants, be nontoxic for the host cells and not interfere with experiments, none of the available antibiotics meet the mentioned criteria. Therefore, application of antibiotics in cell culture should be limited. Instead, a good aseptic practice plays an important role in prevention of contamination.

Microbial contamination in cell culture which is antibiotic free is detectable by turbidity or color changes in cell culture medium. In the case of using antibiotics in a cell culture, there are four possibilities: 1. susceptibility to antibiotics, 2. resistance to antibiotics, 3. partial resistance to antibiotics, 4. resistance to antibiotics only by mycoplasma. The last one is the worst contamination since mycoplasmas can be spread by aerosols. In the case of antibiotic susceptibility, antibiotics prevent the cultivation of bacteria and fungi, but are incapable of precluding mycoplasma from the beginning. Therefore, continuous use of antibiotics for a long time in cell cultures not only is not helpful, but also can cause more problems. However, the use of antibiotics (Penicillin/ Streptomycin) for a short term (the first two weeks) in primary culture is vital. Since antibiotics are unstable in the medium, it is highly suggested to replace antibiotic containing medium with fresh medium every two or three days .

Discard or treat mycoplasma contaminated cells

In the case of mycoplasma contamination of cells which are really valuable and rare, it is suggested to treat them to eliminate mycoplasma infection. Otherwise, it is recommended to discard mycoplasma contaminated cells since they are considered as a source of contamination in the lab .