System:
MycoFog
Mycoplasma contamination poses a significant challenge in cell culture laboratories, compromising research integrity and leading to inaccurate experimental results.
Estimates for the incidence of Mycoplasma contamination vary from 10% to 36% of cell lines used in laboratory procedures. Laboratory incubators, being a critical environment for cell growth, are prone to Mycoplasma contamination.
Explore the effective strategies to eliminate Mycoplasma contamination from laboratory incubators, ensuring reliable and consistent cell culture experiments.
1. Cleaning and Decontamination Procedures
To eliminate Mycoplasma contamination, thorough cleaning and decontamination protocols are essential. The following steps can be employed:
- Regular Cleaning
Implement a stringent cleaning regimen for laboratory incubators, involving the removal of all cell culture vessels, shelves, and trays. Clean surfaces using appropriate disinfectants, such as 70% ethanol or other proven disinfectants, to effectively eliminate mycoplasma.
- Disinfection
After cleaning, disinfect the incubator using (eg) heat treatment, Hydrogen Peroxide Vapor (VHP), Paracetic Acid (PAA), or Ultraviolet (UV) light. These methods are highly effective in eradicating mycoplasma and other contaminants.
- Periodic Maintenance
Establish a maintenance schedule for incubators to ensure proper functioning and prevent Mycoplasma contamination. This includes regular replacement of filters, inspection of seals, and calibration of temperature and humidity controls.
2. Isolation and Quarantine
Preventing mycoplasma contamination requires isolating potentially contaminated cell lines and practicing strict quarantine measures:
- Isolation
Identify and segregate Mycoplasma-contaminated cell lines from healthy ones. Store contaminated cultures separately, preferably in sealed containers, to prevent cross-contamination.
- Quarantine
Newly obtained or suspected cell lines should be quarantined for mycoplasma testing before integration into the laboratory. Maintain a dedicated space for quarantine and perform regular testing to ensure early detection and prompt elimination of contamination sources.
- In some cases it may be possible to recover contaminated cultures using fluoroquinolone antibiotics. If there is no other source for uncontaminated cells, the time and expense may be justified.
3. Regular Testing
Preventing mycoplasma contamination requires isolating potentially contaminated cell lines and practicing strict quarantine measures:
- Testing Methods
Employ validated Mycoplasma detection methods, such as polymerase chain reaction (PCR) or DNA staining assays. These tests are highly sensitive and specific, allowing for accurate detection of mycoplasma contamination.
- Frequency
Establish a periodic testing schedule to ensure the timely identification of Mycoplasma contamination. Regular testing, at least once a month, is recommended, especially for high-throughput laboratories with extensive cell culture activities.
- External Testing
Periodically utilize external services or contract laboratories for Mycoplasma testing to obtain an independent and unbiased assessment of incubator and cell culture contamination.
4. What makes tackling Mycoplasma contamination crucial?
Mycoplasma contamination represents a persistent challenge in laboratory incubators and can compromise the validity of cell culture experiments.
By implementing robust cleaning and decontamination procedures, ensuring isolation and quarantine practices, and implementing regular mycoplasma testing, laboratories can significantly reduce the risk of contamination.
Maintaining a vigilant approach towards mycoplasma elimination is crucial for generating reliable and reproducible experimental results, thus promoting the advancement of scientific research.
References
- Drexler H, Uphof CC (2002). In Vitro Cell. Biol. – Animal 38:79-85
- Olarerin-George AO, Hogenesch JB (2015). Nucleic Acids Research 43(5): 2535-2542
- Uphof CC, Denkman Sabine-A, Drexler HG (2012) J. Biomed. Biotechnol. 2012: 267678
- Chat-GPT 4, OpenAI
