Biological Systems: Theory and Innovation

Identification of mycoplasma contamination in cell culture

Tetiana Тkachenko, M. Kokovin, P. Drozd, Svitlana Prylutska
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Abstract

Mycoplasmas are the smallest and simplest prokaryotes that localise to the endosomes of mammalian cells and are quite common contaminants of cell cultures. In this study, we identified mycoplasma infection in a cell culture of the human breast cancer cell line BR using phase-contrast and fluorescence microscopy with 4,6-diamidino-2-phenylindole (DAPI) staining, which has affinity for nucleolar DNA. Breast cancer cells infected with mycoplasma have increased granularity (phase contrast microscopy), and when fluorescence microscopy is used, mycoplasma DNA appears as fluorescent spots concentrated in the cytoplasm of infected cells and outside them. It has been shown that mycoplasma contamination does not affect the number of tumour cells in culture, but may probably affect the course of their physiological and biochemical functions. To treat mycoplasma infection in cell culture, two antibiotics were used: tiamulin (macrolide group) and minocycline (tetracycline group) at a concentration of 5 mM each. The effectiveness of combined antibiotic therapy of mycoplasmas was confirmed by microscopic examination. The proposed scheme of decontamination of breast cancer cell culture with combined antibiotics allows to completely destroy mycoplasma infection in cultured cells

Keywords

cell culture; mycoplasma; contamination; identification

Suggested citation
Тkachenko, Т., Kokovin, M., Drozd, P., & Prylutska, S. (2023). Identification of mycoplasma contamination in cell culture. Biological Systems: Theory and Innovation, 14(2), 76-83. https://doi.org/10.31548/biologiya14(3-4).2023.007
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