Cell culture is the cornerstone of modern biomedical research and biopharmacutical development. From drug discovery to advanced cell therapy, reliable and contamination free cell lines are required for the production of coherent, reproductive consequences. Among many potential contaminants in cell culture, Mycoplasma presents a particularly insidious challenge. Unlike bacteria or fungi, Mycoplasma is small, cell-least-creating organisms that can quietly infiltrate cultures, often become infinite determined by affecting cellular behavior, metabolism and experimental consequences.
Detection of mycoplasma contamination is important to maintain the integrity of early research and ensure the safety of biopharmacutical products. In this article, we detect the required test methods, their benefits, and regular screening should have a standard practice in every laboratory.
What is Mycoplasma and why is it a concern?
This is one of the smallest free living microorganisms, which range from the 0.1 to 0.3 micrometers in size. Their lack of cell wall allows them to undergo standard bacterial filters and make them resistant to many common antibiotics. In cell culture, mycoplasma contamination may arise from contaminated reagents, cross-establishment, or improper decay techniques between cultures.
The effects of Mycoplasma contamination on cell cultures can be severe:
Converted cell growth and morphology
Low protein synthesis and metabolism
Intervention with nucleic acid synthesis
Contracted practical fertility
Possible security risk in biopharmacutical production
Given these risks, early identification and monitoring are important for both research and industrial applications.
Major methods for detecting mycoplasma
Mycoplasma has several reliable ways to detect contamination, offering different levels of each sensitivity, specificity and the turnaround time. The choice of the method depends on the needs of the laboratory and the type of the cell culture.
- Polymerase Chain Reaction (PCR)
PCR is one of the most widely used and the highly sensitive techniques to identify the mycoplasma in cell cultures. This method works by increasing DNA sequences that are unique to mycoplasma species, which are capable of rapid and accurate detection.
Benefit:
Extremely high sensitivity and the specificity
Rapid results, often available within a few hours
Idea:
Special laboratory equipment and trained personnel are required
Living and dead can not differentiate between Mycoplasma cells
PCR is particularly valuable for laboratories and biopharmacutical features, which require quick and reliable screening to ensure the integrity of their cell cultures.
- Culture law
Culture method involves selective if selective if or broth media increases mycoplasma. While this method is considered a standard of gold to confirm viable contamination, it is more time -taking than PCR.
Benefit:
Detects competent Mycoplasma for replication
Other tests are useful for confirmation of positive results
Idea:
Slow turn -round (may take up to 28 days)
Labor-intensive and strict decaying technology requires
- Fluorescent staining
Fluorescent staining methods use colors that bind to the mycoplasma DNA, allowing visualizations under the fluorescence microscope. Common colors include DAPI and Hoechst.
Benefit:
Quick and relatively easy performance
Useful for regular screening
Idea:
PCR less sensitive
A fluorescence microscope requires
If not executed carefully, you can produce false positive or negative
- Enzyme-Linked Immunosorbent Assay (ELISA)
ELISA cell culture can detecting mycoplasma antigen or antibodies in supernatants. While not usually used as PCR or culture methods, Elisa provides an additional screening option for laboratories.
Benefit:
Relatively simple and scalable for many samples
Special equipment is not required beyond standard ELISA readers
Idea:
PCR less sensitive
Not all mycoplasma species can detect
Best Practice for Mycoplasma Testing
To ensure contamination-free cell cultures, laboratories must apply a broad, multi-level approach to mycoplasma testing:
Routine Screening: Conduct regular testing (eg, monthly or quarterly) to detect contamination to prevent the effects on experiments or production.
Use of certified reagents: Use only reagents and serums that are certified as Mycoplasma free to reduce the risk.
Strict decay technology: To prevent cross-pollution, maintain proper laboratory hygiene including sterilization and careful handling.
Multiple detection methods: The combination of PCR with culture or staining methods improves the accuracy of detecting and reduces false negatives.
Conclusion
Mycoplasma contamination cell remains a frequent challenge in culture and biopharmacutical manufacturing. Understanding its effects, adopting PCR, culture, and fluorescent staining, and rigorous best practices, by adopting methods of reliable detections, laboratories can protect both their research results and patient safety.
For organizations seeking trusted expertise in GMP-compliant mycoplasma testing and quality control, Xellera Therapeutics offers advanced solutions that ensure contamination-free cell cultures, supporting the development of safe and effective biopharmaceutical therapies worldwide.
