Mycoplasma: properties, detection and decontamination methods of cell cultures and viral strains (Review)

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Abstract

Mycoplasma contamination of continuous cell cultures and collection viral strains remains a serious problem in the biotechnology industry and experimental research. The frequency of mycoplasma contamination of cultured cell lines and viruses is 15–35%, in some cases up to 80%. Mycoplasmas cause various changes in cultures contaminated by them, up to cell death, have immunomodulatory properties, and affect the yield of certain viruses propagated in cell culture. Mycoplasmas do not have a cell wall, are able to pass through a bacterial filter, have the smallest genome (≈580 kb) among bacteria, and are capable of independent reproduction and existence. These microorganisms are resistant to most antibiotics commonly used in cell culture. Derivative groups of tetracyclines and fluoroquinolones (BM-Cyclin®, Ciprobay®, Baytril®, Plasmocin®, MRA) have shown certain effectiveness in decontaminating viral strains and cell cultures from mycoplasmas. Timely, highly sensitive detection and prevention of mycoplasma infection is of great importance. For routine scanning of mycoplasma infection of continuous cell cultures and viral strains, the methods of indicator cell culture (cytochemical) and polymerase chain reaction (PCR) are recommended, for more accurate – microbiological analysis of mycoplasma colonies on a special medium.

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About the authors

O. A. Leonovich

Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences (Institute of Poliomyelitis)

Author for correspondence.
Email: leonovich_oa@chumakovs.su
Russian Federation, Moscow, 108819

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2. Fig. 1. Structural diagram of mycoplasma. The presence of a lipoprotein membrane with soluble contents in the form of proteins, RNA, DNA and the absence of a cell wall and nuclear membrane is shown [18].

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3. Fig. 2. (a) Transmission electron microscopy of Mycoplasma agalactiae showing smooth cells and irregular spots of mycoplasma [41]. (b) Mycoplasma colonies on agar showing the typical fried egg shape on solid media, showing the apical structure by which mycoplasmas attach to the host cell.

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4. Fig. 3. Staining of cell culture with Hoechst 33258 [42]. The sample was incubated for 3 days with a pure indicator culture, the cells were fixed, stained with Hoechst 33258 and observed under a fluorescence microscope (×40): (a) Vero cell culture without mycoplasma; (b) rounded nuclei of Vero cells and mycoplasma are visible as separate small hierarchical spots.

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