How to keep mycoplasma away from your cells

According to a rough estimate, about one-third of cell cultures are infected with Mycoplasma. These little things invade your cells without knowing it. Because of their small size and slow growth, they are difficult to detect. However, their presence does change the metabolic process of the cell, which in turn interferes with your experimental results or causes unexplained differences. To make matters worse, it is almost impossible to rescue the infected cells. Therefore, as far as mycoplasma contamination is concerned, prevention is the first priority. Here are some simple precautions that may be useful to you.

Cell source is the key

One of the most common causes of mycoplasma contamination is that the introduced culture has been infected. Once in the laboratory, these little things will spread to every corner and make you collapse. To avoid this, the easiest way is not to request cells from other laboratories, but to purchase them from suppliers or strain collection centers (ATCC or ECACC). These cells are strictly tested to ensure they do not contain mycoplasma.

If it is difficult to purchase, or if you have to use cells from other laboratories for other reasons, you should also be vigilant, isolate them in a separate incubator, and test for mycoplasma at the same time. There are many companies on the market that provide mycoplasma detection kits, you can choose.

Regular testing is important

Even if there are no new cells in the laboratory, mycoplasma may be transferred to the existing culture by the operator. Of course, the operator does not know. Another route for mycoplasma contamination is through your cell culture reagents. Commonly used reagents such as culture media and enzymes may also be contaminated with mycoplasma.

Although the contamination of people or reagents cannot be completely avoided, routine testing can also avoid the deterioration of the situation. Perhaps, when mycoplasma is first infected, we may not be able to detect it, but early detection can prevent the spread of this microorganism and avoid large losses.

There are many types of mycoplasma detection. Although the direct cultivation method has high accuracy, it takes three to four weeks. Therefore, it is enough for us to use indirect detection methods in routine detection. If a positive result is found, a more cumbersome direct detection can be used later. Common indirect detection methods include fluorescence and biochemical analysis based on enzyme activity, PCR analysis, immunoassay, etc.

Aseptic operation should be done

Another major source of mycoplasma contamination is poor aseptic operation. Although good practice may be the simplest precaution, it is easier said than done. Everyone makes mistakes, especially when repeating labor. Therefore, the laboratory should carry out training and supervision from time to time to make everyone fully aware of the destructive nature of mycoplasma.

Many laboratories still use antibiotics, but cell culture personnel gradually realize that this is irresponsible because it brings about the problem of resistant microorganisms. On the contrary, it is best to rely on your own sterile operation to prevent mycoplasma, and only use antibiotics when you suspect or confirm infection.

Preserve the cell in case

Finally, reserve some cells by cryopreservation to prevent large-scale mycoplasma attacks. If there are signs of mycoplasma looking up, or if the routine test is positive, the most reliable remedy is to throw away all cultures, clean the incubator and ultra-clean bench, and start from scratch. Of course, you have to make sure that the stored cells are free of mycoplasma.

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