Identification of mycoplasma contamination in cell culture
DOI:
https://doi.org/10.31548/biologiya14(3-4).2023.007Keywords:
cell culture, mycoplasma, contamination, identification.Abstract
Mycoplasmas are the smallest and simplest prokaryotes wich were found in the endosomes of mammalian cells. They are widespread contaminants in cell cultures. It was identified a mycoplasma infection in a human breast cancer cell line. It was used fluorescence microscopy and nuclear affinity 4,6-diamidino-2-phenylindole (DAPI) staining. For the treatment of mycoplasma infection in cell culture, two antibiotics of the macrolide series (Tiamulin) and tetracyclines (Minocycline) was used. The effectiveness of combined antibiotic therapy against mycoplasmas has been proven, which was confirmed by the microscopic method. Therefore, treatment with combined antibiotics can completely eradicate mycoplasma infection from cultured cells.References
Ahangaran, S., Pourbakhsh, S. A., Abtin, A., & Asli, E. (2019). Isolation and Detection of Mycoplasma pneumoniae from Cell Culture by Culture and PCR. Iranian Journal of Medical Microbiology, 13(3), 153-163.
Borup-Christensen, P., Erb, K., & Jensenius, J. C. (1988). Curing human hybridomas infected with Mycoplasma hyorhinis. Journal of immunological methods, 110(2), 237–240. https://doi.org/10.1016/0022-1759(88)90109-3
Cando‐Dumancela, C., Liddicoat, C., McLeod, D., Young, J.M., & Breed, M.F. (2021). A guide to minimize contamination issues in microbiome restoration studies. Restoration Ecology, 29. https://doi.org/10.1111/rec.13358.
Carthew, J., Abdelmaksoud, H., Cowley, K., Hodgson‐Garms, M., Elnathan, R., Spatz, J., Brugger, J., Thissen, H., Simpson, K., Voelcker, N., Frith, J., & V. Cadarso. (2021). Next Generation Cell Culture Tools Featuring Micro‐ and Nanotopographies for Biological Screening. Advanced Functional Materials, 32(3): 2100881. https://doi.org/10.1002/adfm.202100881.
Coté R. (2001). Assessing and controlling microbial contamination in cell cultures. Current protocols in cell biology, Chapter 1. https://doi.org/10.1002/0471143030.cb0105s01.
Doyle, M., Vodstrcil, L. A., Plummer, E. L., Aguirre, I., Fairley, C. K., & Bradshaw, C. S. (2020). Nonquinolone Options for the Treatment of Mycoplasma genitalium in the Era of Increased Resistance. Open forum infectious diseases, 7(8), ofaa291. https://doi.org/10.1093/ofid/ofaa291
Fay, M.F. (1992). Conservation of rare and endangered plants using in vitro methods, Plant, 28, 1–4. https://doi.org/10.1007/BF02632183
Jung, H., Wang, S. Y., Yang, I. W., Hsueh, D. W., Yang, W. J., Wang, T. H., & Wang, H. S. (2003). Detection and treatment of mycoplasma contamination in cultured cells. Chang Gung medical journal, 26(4), 250–258.
Lawson-Ferreira, R., Santiago, M. A., Chometon, T. Q., Costa, V. A., Silva, S. A., Bertho, A. L., & de Filippis, I. (2021). Flow-Cytometric Method for Viability Analysis of Mycoplasma gallisepticum and Other Cell-Culture-Contaminant Mollicutes. Current microbiology, 78(1), 67–77. https://doi.org/10.1007/s00284-020-02255-1
Ligasová, A., Vydržalová, M., Buriánová, R., Brůčková, L., Večeřová, R., Janošťáková, A., & Koberna, K. (2019). A New Sensitive Method for the Detection of Mycoplasmas Using Fluorescence Microscopy. Cells, 8(12), 1510. https://doi.org/10.3390/cells8121510
McNerney, M. P., Doiron, K. E., Ng, T. L., Chang, T. Z., & Silver, P. A. (2021). Theranostic cells: emerging clinical applications of synthetic biology. Nature reviews. Genetics, 22(11), 730–746. https://doi.org/10.1038/s41576-021-00383-3
Nikfarjam, L., & Farzaneh, P. (2012). Prevention and detection of Mycoplasma contamination in cell culture. Cell journal, 13(4), 203–212.
Pastore, M., Marcone, C., Pennone, F., Cristinzio, G., & Ragozzino, A. (1995). Detection of Mycoplasma-like organisms (MLOs) in apricot by fluorescence microscopy (DAPI) in the South of Italy. Acta Horticulturae, 386, 506-510.
Peng, W., Datta, P., Ayan, B., Ozbolat, V., Sosnoski, D., & Ozbolat, I. T. (2017). 3D bioprinting for drug discovery and development in pharmaceutics. Acta biomaterialia, 57, 26–46. https://doi.org/10.1016/j.actbio.2017.05.025
Regent, F., Morizur, L., Lesueur, L., Habeler, W., Plancheron, A., Ben M'Barek, K., & Monville, C. (2019). Automation of human pluripotent stem cell differentiation toward retinal pigment epithelial cells for large-scale productions. Scientific reports, 9(1), 10646. https://doi.org/10.1038/s41598-019-47123-6
Strober W. (2015). Trypan Blue Exclusion Test of Cell Viability. Current protocols in immunology, 111, A3.B.1–A3.B.3. https://doi.org/10.1002/0471142735.ima03bs111
Timenetsky, J., Santos, L. M., Buzinhani, M., & Mettifogo, E. (2006). Detection of multiple mycoplasma infection in cell cultures by PCR. Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas, 39(7), 907–914. https://doi.org/10.1590/s0100-879x2006000700009
Uphoff, C. C., & Drexler, H. G. (2014). Detection of Mycoplasma contamination in cell cultures. Current protocols in molecular biology, 106, 28.4.1–28.4.14. https://doi.org/10.1002/0471142727.mb2804s106
Uphoff, C. C., Denkmann, S. A., & Drexler, H. G. (2012). Treatment of mycoplasma contamination in cell cultures with Plasmocin. Journal of biomedicine & biotechnology, 2012, 267678. https://doi.org/10.1155/2012/267678
Uphoff, C. C., Gignac, S. M., & Drexler, H. G. (1992). Mycoplasma contamination in human leukemia cell lines. I. Comparison of various detection methods. Journal of immunological methods, 149(1), 43–53. https://doi.org/10.1016/s0022-1759(12)80047-0
Uphoff, C. C., Meyer, C., & Drexler, H. G. (2002). Elimination of mycoplasma from leukemia-lymphoma cell lines using antibiotics. Leukemia, 16(2), 284–288. https://doi.org/10.1038/sj.leu.2402364.
Uphoff, C.C., Drexler, H.G. (2013). Detection of Mycoplasma Contaminations. In: Helgason, C., Miller, C. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology, vol 946. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-128-8_1
Work T.S., Work E., Adams R.L.P., & Burdon R.H. (1980). Laboratory Techniques in Biochemistry and Molecular Biology: Cell culture for biochemists. Vol. 8, New York: North-Holland Publishing Company.
Yin, Z. F., Zhang, Y. N., Liang, S. F., Zhao, S. S., Du, J., & Cheng, B. B. (2019). Mycoplasma contamination-mediated attenuation of plasmid DNA transfection efficiency is augmented via L-arginine deprivation in HEK-293 cells. Journal of Zhejiang University. Science. B, 20(12), 1021–1026. https://doi.org/10.1631/jzus.B1900380
Young, L., Sung, J., Stacey, G., & Masters, J. R. (2010). Detection of Mycoplasma in cell cultures. Nature protocols, 5(5), 929–934. https://doi.org/10.1038/nprot.2010.43
Yu, T., Wang, Y., Zhang, H., Johnson, C. H., Jiang, Y., Li, X., Wu, Z., Liu, T., Krausz, K. W., Yu, A., Gonzalez, F. J., Huang, M., & Bi, H. (2016). Metabolomics reveals mycoplasma contamination interferes with the metabolism of PANC-1 cells. Analytical and bioanalytical chemistry, 408(16), 4267–4273. https://doi.org/10.1007/s00216-016-9525-9
Prylutskyi Yu.I., Ilchenko O.V., Tsymbaliuk O.V., & Kosterin S.O. (2017). Statystychni metody v biolohii, Kyiv : Nauk. dumka.
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