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If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate. Advanced search. Skip to main content Thank you for visiting nature. Subjects Bacteria Cell culture. Abstract Mycoplasma is a prokaryotic organism that is a frequent and occult contaminant of cell cultures. Access through your institution. Buy or subscribe.
Rent or Buy article Get time limited or full article access on ReadCube. Figure 1. Figure 2: Typical gel photo from direct PCR. References 1 Hay, R. View author publications. Ethics declarations Competing interests The authors declare no competing financial interests. Rights and permissions Reprints and Permissions. About this article Cite this article Young, L. Copy to clipboard. Lamson , Adrian Berger , Katherine C.
Tafreshian , Bernardo Tavora , Roger D. Mycoplasma was first detected in cell culture in , so why is it still such an issue? Detection is one challenge, but it is also more difficult to prevent and eliminate than other types of contaminants. It has a pliable membrane that makes it resistant to pressure, temperature, osmolality and dehydration.
There are over different species of mycoplasma with over 20 found as contaminants in cell culture. Mycoplasma can infect all types of eukarayotic cells. They have limited metabolic capabilities and they adapt and thrive in a cell culture environment. Because it often goes undetected, research continues to go on with contaminated cells and these cells are often shared with other labs, where contamination can spread.
This inability to reproduce results brings research results into question and can negate countless hours of work and research dollars. The most common mycoplasma cell culture contaminates are human, bovine or swine in origin, with human being the largest. During the webinar, some interesting statistics were presented on how humans introduce mycoplasma that I wanted to share. First, In addition to mycoplasma contamination through human introduction, it can also be introduced through cross contamination with an already infected culture.
Poor lab techniques are at fault for much of the cross contamination. Things like using the same media bottles, reusing pipette tips or poor sterile technique can spread mycoplasma from one culture to another. The next part of the webinar, I found extremely informative with great tips on how to prevent a mycoplasma contamination from happening in the lab. One way to help keep the lab clean and to prevent opportunities for contamination is through the use of a Pharmacidal Spray.
Pharmacidal Spray offers a powerful prevention solution for disinfecting and sterilizing benchtops and equipment.
They prevent the growth of common contaminants such as bacteria, fungi, viruses and mycoplasma. It is more powerful than ethanol and other common lab alcohols for disinfecting benchtops and equipment, yet safe enough even to use on bare hands. Pharmacidal Spray is non-toxic and biodegradable.
It is also non-corrosive and can be used on all common work surfaces. Also recommended is cleaning the incubator water pan regularly. Another aid in preventing contamination in these areas is to use a preventative solution that is added to the water. The Aquaguard solutions are highly active, non-volatile, non-corrosive, and non-toxic treatment for maintaining water quality and preventing contamination in the water pan of a cell culture incubator.
They have been validated for use in incubators and water baths actively culturing sensitive stem cells. Aquaguard 1 Solution, when used as directed, have been shown to have no effect on the morphology, cell proliferation and expansion, pluripotency marker expression, or karyotype of human pluripotent stem cells.
Contamination of a cell culture by mycoplasmas cannot be visualized, as it does not generate the turbidity typically associated with bacterial or fungal contamination.
Furthermore, the consequent morphological changes and altered growth rates in affected cell cultures can be minimal or simply unapparent. Mycoplasmas compete with host cells for biosynthetic precursors and nutrients and can alter DNA, RNA and protein synthesis, diminish amino acid and ATP levels, introduce chromosomal alterations, and modify host-cell plasma membrane antigens.
A microarray analysis on contaminated cultured human cells has revealed the severe effects that mycoplasmas can have on the expression of hundreds of genes, including some that encode receptors, ion channels, growth factors and oncogenes [4].
Moreover, mycoplasmas exert significant effects on cultured immune cells such as monocytes and macrophages. Mycoplasmas contain highly immunogenic lipoproteins anchored on the outer face of the plasma membrane. These lipoproteins are recognized by specific pattern recognition receptors on immune cells—in particular, Toll-like receptor 2 TLR2. Upon recognition of mycoplasmal lipoproteins, TLR2 induces the NF-kB pathway, which leads to activation of these cells and consequently, to biased experimental results.
There are three major sources leading to mycoplasma contamination of cell cultures in the laboratory: infected cells sent from another lab; contaminated cell culture medium reagents such as serum and trypsin; and laboratory personnel infected with M. Furthermore, contamination can spread rapidly to other cell lines through dispersion of aerosol droplets. Once mycoplasmas have been detected, the best solution to eliminate them and to prevent them from spreading is to discard the contaminated cell line.
However, valuable cell lines that are too precious to be sacrificed can be salvaged by treatment with effective mycoplasma-eradication products, including mycoplasma-selective antibiotics. These products have been shown to eliminate mycoplasma and to restore cell behavior and responses within days or weeks after treatment [5].
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