How did the rate of cell proliferation become so slow? What happens when the cells become diseased, the cells become rounded, and they fall off the walls of the flasks? It's crazy, why is it so hard to grow cells that exist during the experiment"Ghost"., even seasoned veterans have to face, yes, it isMycoplasma infection
The threat of mycoplasma infection
MycoplasmaThe infection rate is as high as 63%.This also makes the problem of "mycoplasma contamination" in the cell culture process global, and researchers are often confused when it comes to mycoplasma contamination.
It is a prokaryotic microorganism that resembles bacteria but does not have a cell wall, with a diameter of 50-300 nm and can easily pass 0The 22um filter membrane is mixed into the culture system, and ordinary antibiotics (such as bispecific antibodies, commonly used in cultured cells) do not work on it.
Moreover, cell cultures often have no obvious signs of mycoplasma contamination, unlike bacterial or fungal contamination that has visible changes to the naked eye, and even the cells themselves can continue to maintain normal morphology and proliferation rates for a period of time.
These characteristics have brought us some trouble in judging and dealing with mycoplasma contamination.
PossiblyThis leads to inaccurate experimental resultsBecause mycoplasma interacts with other components in the medium, affecting the growth of other microorganisms, thus interfering with the collection and analysis of experimental results.
Mycoplasma contamination may have a positive effect onExperimental equipment and instruments caused damage, especially for biosafety cabinets and other related equipment in the laboratory, as mycoplasma can grow on them and form a bacterial film, affecting the normal operation of the equipment. If the number of mycoplasma is low, it may not have a noticeable effect on the sequencing results.
If mycoplasma are in large numbers, they may affect the quality of the sample during sequencing, thuscausedSequencing results are inaccurate。In addition, mycoplasma contamination may result in a decrease in the number of sequence reads in the sample, which can affect subsequent analysis results such as sequence assembly and gene expression analysis.
Therefore, appropriate measures should be taken to avoid and reduce mycoplasma contamination before sequencing, such as the use of aseptic techniques, strict control of the laboratory environment, and the use of high-quality reagents and instruments. If mycoplasma contamination has already occurred, it should be detected and treated promptly to reduce the impact on sequencing results.
Mycoplasma detection modality
We are not at a loss for this, and through strict disinfection measures and reasonable laboratory management, we can effectively prevent mycoplasma infection and ensure the smooth progress of research. In view of the important negative impact of mycoplasma contamination on cell biology experiments, regular testing of cells for mycoplasma contamination has become an inevitable option for many laboratories.
Common mycoplasma detection techniques include: isolation and culture assays, DNA fluorescent staining assays, etc., but they are more recommended hereIsothermal amplification。Among them, the isolation culture method and the DNA staining method should be more difficult to operate, whileThe isothermal amplification method has low concentration, low cytotoxicity, and is easy to use, ready to use, as long as it is added to mycoplasma-contaminated cultured cells for one week of incubation