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Cell Culture Contamination Control

Cell Culture Contamination Control: What Are Mycoplasma?

The Stealthy Invader





What Are Mycoplasma and Why Are They so Feared?


Mar 26, 2024 | Informative Articles


 

 Featured in this article:

• What is mycoplasma contamination?
• What happens when a cell culture is contaminated by mycoplasma?


In the world of microbiology, there exist tiny organisms causing big trouble – mycoplasma.
Mycoplasma are the smallest prokaryotic organisms and the most common contaminants in cell cultures. They can easily pass through sterile filtering systems and are resistant to commonly used antibiotics. Mycoplasma contamination is a serious problem for academic and biopharmaceutical laboratories due to its impact on cell properties and safety of biological products. A contamination often remains undetected and pose significant challenges to cell culture experiments, potentially comprising data integrity and research outcomes. It is estimated that mycoplasma are present in almost every lab and that about 5 to 30 % of the world’s cell lines are contaminated with mycoplasma, causing enormous costs and effort [1].


In this article we give a brief overview of mycoplasma contamination. Learn about the biology of mycoplasma and the consequences of a mycoplasma contamination to understand how cells are contaminated.

What is Mycoplasma Contamination?  The Biology of Mycoplasma

 
Mycoplasma Structure

Mycoplasma are the smallest self-replicating bacteria (about 0.15 - 0.3 µm) and belong to the class of mollicute [1]. In fact, mycoplasma and mollicutes are commonly used as synonyms. Unlike other bacteria they lack a cell wall, makes them capable of assuming various shapes and resistant to commonly used antibiotics. Due to their flexible membrane and small size mycoplasma are able to pass through filtering systems.

They cannot be detected visually - neither by the naked eye nor by microscope. Even highly contaminated cell cultures do not show any turbidity or other obvious symptoms.

What makes mycoplasma so special?

• Absence of cell wall
• Flexible membrane
• Invisible
• Pass through filters
• Resistant to common antibiotics

 

 

Mycoplasma lack a cell wall, making them resistant to commonly used antibiotics. The flexible membrane allowing them to shrink to diameters < 0.45 µm and pass through filtering systems.

 

What happens when a cell culture is contaminated by mycoplasma?  Consequences of a mycoplasma contamination

Mycoplasma species are known to infect a variety of eukaryotic cells, including human, animal and plant cells.

When a cell culture becomes infected with mycoplasma, it can lead to various detrimental effects on the cells and their growth. Mycoplasma, being simple parasitic organisms, rely on the host cells for their metabolic needs. Initially, they attach to the host cell and then can merge with its membrane. Following this, mycoplasma replicate rapidly, overgrowing significantly the host cells.

Unlike other bacterial contaminants, mycoplasma contamination doesn't typically cause visible changes in the growth medium. However, its impact on cell cultures can be profound and can lead to:

• Altered cell metabolism
• Inhibition of cell growth and proliferation
• Reduced transfection efficiency
• Changes in gene and protein expression
• Chromosomal abnormalities
• Impair nucleic acid synthesis
• Apoptosis

 

These effects can vary depending on the cell line and the species of mycoplasma involved. Nevertheless, mycoplasma contamination poses a serious challenge to cell culture research, as it can compromise the reliability, reproducibility and validity of experimental results.

The presence of mycoplasma in cell culture has different side effects, including:

• Loss of time
• Loss of money
• Loss of valuable cells
• Potential dissemination of inaccurate findings through publications
• Misleading publications
• Biosafety concerns

If a contamination remains undetected, contaminated cultures can endanger scientific data, delay research projects and jeopardise the safety of biopharmaceutical products. Therefore, it's crucial for researchers to be vigilant in detecting and preventing mycoplasma contamination in cell cultures.

The impact of mycoplasma contamination in cell culture can be profound and represent a serious problem for academic and biopharmaceutical laboratories due to its impact on cell properties and safety of biological products.

 

References

[1] Nikfarjam L, Farzaneh P. Prevention and detection of Mycoplasma contamination in cell culture. Cell J. 2012 Winter;13(4):203-12. Epub 2011 Dec 22. PMID: 23508237; PMCID: PMC3584481.

Mycoplasma Standards 100GC®

100GC® Mycoplasma Standards are non-infectious reference materials for matrix-specific validation of PCR-based mycoplasma tests

Minerva Biolabs

Catalogue No.DescriptionPack SizePriceQty
104-1001100GC® Mycoplasma StandardMycoplasma arginini Standard £355.00 Quantity Add to Order
104-2001100GC® Mycoplasma StandardMycoplasma orale Standard £355.00 Quantity Add to Order
104-2001100GC® Mycoplasma StandardMycoplasma orale Standard £355.00 Quantity Add to Order
104-2001100GC® Mycoplasma StandardMycoplasma orale Standard £355.00 Quantity Add to Order
104-2001100GC® Mycoplasma StandardMycoplasma orale Standard £355.00 Quantity Add to Order
104-2001100GC® Mycoplasma StandardMycoplasma orale Standard £355.00 Quantity Add to Order
104-2001100GC® Mycoplasma StandardMycoplasma orale Standard £355.00 Quantity Add to Order
104-2001100GC® Mycoplasma StandardMycoplasma orale Standard £355.00 Quantity Add to Order
104-9001100GC® Mycoplasma StandardMycoplasma citri Standard £355.00 Quantity Add to Order
104-9001100GC® Mycoplasma StandardMycoplasma citri Standard £355.00 Quantity Add to Order

Description

100GC® Mycoplasma Standards are non-infectious reference materials for matrix-specific validation of PCR-based mycoplasma tests.
- Compliant with EP 2.6.7 (12.2)
- Ready-to-use 100 GC/ml standards for reliable matrix-specific validation
- Non-infectious and regulatory safe

Mycoplasmas represent one of the greatest contamination risks in cell cultures and in biopharmaceutical production (e.g. ATMPs). European Pharmacopoeia (EP) 2.6.7 requires a validated test method for the release of such products, capable of detecting both culturable and non-culturable mycoplasmas. Nucleic acid amplification techniques (NAT) have become the new gold standard, as they reliably detect both forms. The previously applied sensitivity limit of 10 CFU/ml was based on classical culture methods.

With the new edition 12.2 of EP 2.6.7, the requirements change: For PCR methods, a sensitivity limit of less than 100 genome copies/ml (GC/ml) is introduced. This PCR-appropriate unit is considered practical for PCR laboratories and has long been expected as a standard for determining the detection limit.

Our 100GC® Mycoplasma Standards are specifically designed to meet these new requirements. They are non-infectious and therefore safe to handle, titrated to 100 GC, and comply with the new regulatory guideline for NAT standards. The standards are lyophilized and ready to use after reconstitution. In addition, the standards can be used as an external positive control with defined concentration, fulfilling all new regulatory requirements.

Optionally, 10 CFU/ml standards remain available for comparability with classical culture methods.

Application Mycoplasma detection
Use cases
  • Method validation according to EP 2.6.7
  • External positive control for mycoplasma testing
  • QC batch release and routine testing
Contents
  • 3 vials each containing 100 GC (lyophilized)
  • 1 vial PCR-grade water
Detection method
  • PCR
  • real-time PCR (qPCR)
  • digital PCR (dPCR/ddPCR™)
Compliance EP 2.6.7
Required equipment
& consumables
  • PCR thermal cycler
  • PCR reaction tubes
  • Centrifuge for 1.5 mL PCR tubes
  • Pipettes
  • Mycoplasma detection kit
Concentration after
reconstitution
100 GC/ml
Sample type RNA-containing DNA standard
Storage 2-8 °C
Shipping Ambient temperature
Shelf life See expiration date on package

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200