Understanding the Basics of Mycoplasma Contamination in Cell Culture
Mycoplasma contamination is a significant problem in cell culture research. It is estimated that over 30% of all cell lines may be contaminated with mycoplasma, affecting results and wasting time and resources. Moreover, mycoplasma contamination can also affect the reproducibility, reliability, and validity of experiments.
In this article, we will walk you through the basics of mycoplasma contamination in cell culture, including its causes, symptoms, detection, and prevention strategies.
What are Mycoplasmas?
Mycoplasmas are among the most common bacterial contaminants that can affect cell cultures. They are small, gram-negative bacteria without a cell wall, making them difficult to detect and treat. Mycoplasmas are ubiquitous in nature and can contaminate cell culture from various sources, including laboratory personnel, animal sources, and contaminated cell culture reagents.
Causes and Symptoms of Mycoplasma Contamination
Mycoplasma contamination can occur at any stage of cell culture, from the moment cells are isolated to the final assay. The most common cause of mycoplasma contamination is the introduction of contaminated reagents, such as fetal bovine serum, antibiotics, or enzymes, or cross-contamination from contaminated cell lines or the environment.
The symptoms of mycoplasma contamination can vary, but common signs include slow or abnormal cell growth, cell death, changes in cell morphology, chromosome abnormalities, and reduced transfection efficiency. These symptoms can affect the accuracy, reliability, and reproducibility of experimental results and can lead to misinterpretation and false conclusions.
Detecting Mycoplasma Contamination
Mycoplasma contamination is often difficult to detect because mycoplasmas are small and do not produce visible signs of contamination. Moreover, mycoplasmas can suppress the immune response of the host cells, making it challenging to detect their presence.
There are several methods for mycoplasma detection, including PCR-based assays, DNA staining, and microbial culture. PCR-based assays are sensitive, specific, and rapid, but they require specialized equipment and expertise. DNA staining is cheap and straightforward, but it lacks specificity and sensitivity. Microbial culture is specific but can take up to a month to obtain results.
Preventing Mycoplasma Contamination
Preventing mycoplasma contamination is essential for successful cell culture research. Several strategies can be employed to reduce the risk of mycoplasma contamination, including strict adherence to aseptic techniques, the use of certified and tested reagents, and the periodic testing of cell lines for mycoplasma contamination. Moreover, regular education and training of laboratory personnel in proper cell culture techniques and mycoplasma detection and prevention can help reduce the incidence of mycoplasma contamination.
Conclusion
In conclusion, mycoplasma contamination is a pervasive problem in cell culture research that can affect the accuracy, reliability, and reproducibility of experimental results. Detecting and preventing mycoplasma contamination is essential for successful cell culture research. Researchers must take proactive steps to reduce the risk of mycoplasma contamination, including strict adherence to aseptic techniques, regular testing of cell lines for mycoplasma contamination, and regular education and training of laboratory personnel.
