Mycoplasma is a serious threat to cell biologists as it can interfere with cell metabolism and affect cell viability. Once contaminated, mycoplasma spreads extremely rapidly and can ultimately ruin the entire culture in the lab. Lack of visual detection makes it extremely difficult to detect mycoplasmas in culture and it may be too late before we notice some deleterious effects! Neglected as a quality control measure, regular mycoplasma testing is indispensable for maintaining high-quality culture conditions. There are a variety of testing methods available commercially including PCR based, DNA based and culture methods for detection. Of these, PCR based methods are highly sensitive and can detect target levels in culture supernatant through quick testing. They can cover a diverse range of mycoplasma strains. Moreover, they are inexpensive kits utilizing agarose gel-based detection endpoints for confirmation. The readily available kits are packed with positive and negative controls for validation.
Eliminating mycoplasma- A Herculean task!
Removal of mycoplasma is a tedious task and is often impossible to achieve. While it is recommended to discard entire cultures infected with mycoplasma, it may not always be possible with sensitive cultures running in the lab. This further reinforces the need to maintain a high level of sterility and routine testing to avoid any chance of infection. An inexpensive method for elimination is the use of a combination of strong antibiotics. These act by interfering directly with protein synthesis or DNA replication. However, the downside to their use is the long treatment duration and high chances of development of resistance. Moreover, the antibiotics could have toxic effects on the cells and interfere with their physiological processes during the long course of treatment. This increases chances of re-infection of the cultures with mycoplasma. Again, depending on the strain of mycoplasma, the treatment regime may vary and become more complicated. Identifying the source of mycoplasma is to prevent any future infection.
Prevention of infection
One of the best practices that could be followed in procurement of high quality, certified cell cultures for all research applications. Cell lines must further be tested for all contaminants upon receipt to validate quality. In fact, any incoming cell culture from outside the laboratory should be subjected to rigorous testing and should be discarded immediately upon detection of contaminants. Kosheeka is a supplier of standard primary cells that have undergone rigorous testing and quality control measures to ensure high quality to customers. It is committed to providing cells that are safe to ensure a smooth user experience. Good laboratory practices and strict implementation of Standard Operating Practices (SOPs) in the lab will enforce the necessary discipline among the users of the culture laboratory. It is also best to work with one cell line at a time since aerosol is the fastest route of spread of any infection. All culture reagents must be separated for different cell lines to avoid cross-contamination. Finally, laboratory personnel are a huge source of contamination and adopting sterile practices such as regular disinfection of the work area and equipment could play a huge role in prevention of mycoplasma.